Invited Review ANIMAL MODELS OF HUMAN LUNG DISEASE
Animal models of chronic obstructive pulmonary disease
Joanne L.Wright,1Manuel Cosio,2and Andrew Churg 1
1Department of Pathology,University of British Columbia,Vancouver,British Columbia;and 2Respiratory Division,Royal
Victoria Hospital,and Meakins-Christie Laboratories,McGill University,Montreal,Quebec,Canada
Wright JL,Cosio M,Churg A.Animal models of chronic obstructive pulmo-
nary disease.Am J Physiol Lung Cell Mol Physiol 295:L1–L15,2008.First
published May 2,2008;doi:10.1152/ajplung.90200.2008.—The mechanisms in-
volved in the genesis of chronic obstructive pulmonary disease (COPD)are poorly
de?ned.This area is complicated and dif?cult to model because COPD consists of
four separate anatomic lesions (emphysema,small airway remodeling,pulmonary
hypertension,and chronic bronchitis)and a functional lesion,acute exacerbation;
moreover,the disease in humans develops over decades.This review discusses the
various animal models that have been used to attempt to recreate human COPD and
the advantages and disadvantages of each.None of the models reproduces the exact
changes seen in humans,but cigarette smoke-induced disease appears to come the
closest,and genetically modi?ed animals also,in some instances,shed light on
processes that appear to play a role.
chronic obstructive pulmonary disease
CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD)is an increas-ingly important cause of morbidity and mortality.COPD is now the ?fth leading cause of death worldwide (109),and recent estimates suggest that the prevalence is as high as 9–10%of adults over age 40(38,131).In the developed world,cigarette smoking is by far the most important risk factor for COPD.Exposure to air pollution particles,occupational expo-sures to dusts and fumes (39),and,in the developing world,exposure to biomass fuels used for cooking is also believed to be etiological agents of COPD (38).There are few animal models of COPD related to air pollu-tion particles,dusts and fumes,and biomass fuels;most COPD models have either used cigarette smoke or other approaches,including genetic modi?cations to mice,believed to reproduce some of the mechanisms behind cigarette smoke.We have recently reviewed mechanistic studies of smoke-exposed ani-mal models (16).In this paper,we will focus on the pros and cons of different COPD animal models.The ?rst question to ask in this context is what would
constitute a perfect model of human COPD.This is not a simple question because human COPD consists of at least four anatomic lesions (further de?ned below):emphysema,small airway remodeling (including goblet cell metaplasia),chronic bronchitis,and pulmonary hypertension;a given patient may have any or all of these lesions.In addition,COPD patients may develop acute exacerbations that are believed to have an infectious basis.To further complicate matters,no matter which lesions are present,COPD develops and is slowly progressive over many years.Because COPD is closely related to the underlying anatomy of the lung,a good animal model should have pulmonary anatomy similar to that of humans.The fundamental mecha-nisms behind COPD should be similar in animal models and humans.The ideal model would allow the investigator to produce the various different anatomic lesions just listed,all in a short period of time.Unfortunately,all of the known animal models meet only some of the above criteria,and even another human would probably not meet all of them,because,as noted above,there is considerable human to human variation in the pattern of COPD.
A variety of different approaches have been used to attempt to model COPD in animals,but there are some more general principles that will affect any COPD model,namely,1)con-siderable differences among species in lung development and maturation;2)differences among species in lung anatomy;and 3)susceptibility to injurious agents among species and strains within a species (discussed in Ref.16).
Comparative Anatomy
Development and maturation.The timing of pre-and post-natal development and maturation of the lung varies widely between species,and this problem needs to be taken into account when planning the initial exposure to any injurious agents.
The rat and mouse have no true alveoli at birth and develop all of their alveoli postnatally,with the majority of alveolo-genesis taking place between days 4and 14(90),whereas guinea pigs are well alveolated at birth and have only a small
increase in numbers of alveoli as they age.Air space size
normally increases with age,but this also varies with species.
For example,guinea pigs (155)and rats (56,96)have a progressive increase in size until 12and 18mo [although there appears to be some differences between strains (56)],and Address for reprint requests and other correspondence:J.L.Wright,Dept.of Pathology,Univ.of British Columbia,2211Wesbrook Mall,Vancouver,B.C.,Canada V6T 2B5(e-mail:jlwright@interchange.ubc.ca).Am J Physiol Lung Cell Mol Physiol 295:L1–L15,2008.
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hamsters attain maximum size by6wk(reviewed in Ref.61). Air space size change also varies among mouse strains,with BALB/c mice demonstrating increasing size up to19mo(61), whereas strain129has constant size after4wk(97).
The importance of recognizing lung maturation and growth patterns cannot be overstated.Administration of an injurious agent in the immediate postnatal period can interfere with alveolarization,and the enlarged air spaces may be falsely interpreted as emphysema.This is also true of genetically modi?ed animals where the modi?cation itself alters lung development in such a way that the animal has larger than normal air spaces(see Genetic and Genetically Modi?ed Mod-els of COPD).
Anatomic structure:bronchial tree.The various animal species commonly used to model COPD differ signi?cantly from each other and from humans in anatomy of the bronchial tree.Mice and rats have less extensive airway branching than do humans(93).Rodents do not normally have respiratory bronchioles;rather,a respiratory bronchiole-like type of meta-plasia is induced by reaction to injury(112).This is consider-ably different from humans,where there are several genera-tions of membranous and respiratory bronchioles.In humans, the membranous,and to a lesser extent,the respiratory bron-chioles,are the targets of smoke-induced small airway remod-eling,whereas the respiratory bronchioles are the initial site of injury in centrilobular emphysema.
Anatomic structure:airway epithelium.In humans,the tra-cheal epithelium is pseudostrati?ed and changes to a simple epithelial type in the subsegmental bronchi(55).The majority of the cells are ciliated and are present throughout the bronchial tree,gradually decreasing in proportion,but extending into the respiratory bronchioles(99).Goblet cells are plentiful in the large airways and progressively decrease until they are rare in the terminal bronchioles(99).This pattern is recapitulated in the guinea pig(Fig.1)(55).
Rats and mice tend to have?10-fold greater numbers of nonciliated bronchiolar cells in the small airways compared with humans(113,166),and the cells vary ultrastructurally among species(113,114).The tracheal epithelium of the mouse is a single columnar layer,whereas in the rat,the transition between pseudostrati?ed and simple epithelium oc-curs at the hilum of the lung.In the mouse,goblet cells are rare in the trachea and almost totally absent in the membranous bronchioles;the majority of the tracheal epithelial cells are nonciliated Clara-like cells(105),although ciliated cells occur in large patches and account for?37%of the total cells(104). In the rat,the proportion of ciliated cells to nonciliated cells increases from upper trachea to bronchioles where they form ?65%of total cells(55).Only a few goblet cells are present in the trachea and large airways and are generally not seen in the small airways.
Anatomic structure:bronchial glands.Interestingly,this appears to be an area of controversy,and this issue is impor-tant,since human chronic bronchitis is characterized by hyper-trophied and hyperplasic bronchial glands and excess mucus secretion.Although most authors agree that the size of the bronchial glands in laboratory animals is smaller than in humans,there is disagreement as to the number and
positioning
https://www.doczj.com/doc/ed6510579.html,parison of tracheal epithelium from humans(A),guinea pigs(B),rats(C),and mice(D).In the human there are large numbers of ciliated cells and
goblet cells,a pattern that is also found in the guinea pig.The rat has scant goblet cells,and the mouse has none.Ciliated cells are common in the rat but are present only in clusters(arrows)in the mouse.Bar?100?m;hematoxylin and eosin(H&E)stain.
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of glands in the various species (32,54,152)(Fig.2).Despite these differences of opinion,it seems clear that bronchial glands are absent in the mouse and rabbit,almost absent in the hamster,present in the rat but concentrated in the upper portion of the trachea,and irregularly distributed in the guinea pig.Even when present,the number of glands in small animals is markedly less than large mammals,thus making small animals a very poor model for diseases such as chronic bronchitis.
Anatomic structure:pulmonary arteries and veins.In hu-
mans,the arterial and venous vasculature have de?nite courses
and are separate from each other:the arteries follow the path of
the airways,whereas the veins are present in the interlobular
septa (Fig.3).The main pulmonary artery and vessels larger
than 1,000?m in diameter are elastic in nature with short,
branching,elastic ?brils arranged in laminae.Vessels between
500and 100?m are muscularized,with a tunica media present
between two elastic laminae.Muscle becomes less apparent
with decreasing size as the spiral of muscle becomes increasing
separated until it disappears at the level of the distal arterioles.
Pulmonary venules have a single elastic lamina and gradually
acquire a muscular layer,the borders of which are generally ill
de?ned as it also contains irregular elastic ?brils.At the level
of the atrium,there may be a few cardiac muscle ?bers.
In the rat,guinea pig,and mouse,interlobular septa are
absent (94).The pulmonary artery branches follow the airways,
but the pulmonary vein is independent and follows a separate course from the periphery of the lung to the hilum.The main and large pulmonary arteries are muscular rather than elastic,
but there is a similar branching pattern and vessel distribution
to that found in the human (45).In the guinea pig (94),the
walls of the small arteries have intermittent areas of increased
muscular thickness.In rodents,there is a wide variation in the
point at which the arterial branches change from muscular to
nonmuscular,with ?50%of vessels adjacent to the terminal
bronchioles and 80%of vessels adjacent to the alveolar ducts
being nonmuscular (45).
The arteries have a dense mesh of ?-smooth muscle actin immunopositive cells which is orga-
nized in a spiral until the most distal vessels (106).Unlike humans,rodent pulmonary veins have a large amount of
cardiac muscle at the hilum of the lung (106),and this is
accompanied by a loose mesh of ?-smooth muscle actin
immunopositive cells,which decreases as the veins become
smaller.
Anatomic structure:bronchial arteries.The bronchial arte-
rial supply is known to be important in the overall lung reaction
to injury,and thus anatomy and distribution are important in
any animal model.There is considerable person-to-person
variation in the human bronchial artery supply (99,135).These
arteries invest the bronchi and extend throughout the length of
the bronchial tree,with communication between the bronchial
and pulmonary circulations in the terminal respiratory unit.In
mice (116),a bronchial artery supply exists,and there are
interconnections between the two vascular systems at the level
of the capillaries.Rats have a relatively similar system to
humans,but with slightly different vascular derivations (135).
Lung Mechanics
A careful quanti?cation of air space size utilizing morphom-
etry should be mandatory to de?ne “emphysema”as a ?rst step
to assess the response to the exposure or manipulation used in
the intended animal model.However,as noted by Henson and Vandivier (44),it is remarkably easy to induce alveolar en-largement with a very wide variety of manipulations in the mouse,the most widely used animal as a model for emphy-sema,and this raises the question of how speci?c such ?ndings might be,particularly since many such manipulations appear to be interfering with lung growth or producing abnormal
lung Fig.3.A lung lobule in a human.Note the venous septa that bound the lobule (arrows)and the airway-artery unit in the center of the lobule.Lobules are not
found in rodents.Bar ?1,000?m;H&E
stain.
https://www.doczj.com/doc/ed6510579.html,parison of bronchial glands between human (A )and guinea pig
(B ).The human gland is large and well formed,whereas the guinea pig gland consists of a few acini only.Bar ?200?m.H&E and alcian blue stain,
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development (see Elastase Emphysema Model and Genetic and Genetically Modi?ed Models of COPD ).Another approach is to use pulmonary function to de?ne emphysema on the basis that,while pulmonary function tests are less sensitive than morphometry and thus may detect only more severe degrees of airways remodeling or parenchymal de-struction,they demonstrate meaningful physiological changes.Although mild emphysema may have normal lung mechanics,more marked emphysematous lung destruction ought to have abnormal lung mechanics.The most speci?c function test representative of emphysema is loss of elastic recoil,deter-
mined from pressure-volume (P-V)curves (increased compli-ance or decreased elastance).This issue is nicely illustrated in the following papers.Foronjy et al.(25)found that long-term cigarette smoke expo-sure in two strains of mice (CBA/JXC57BL/GJ)and A/J resulted in a 17%and 32%increase in mean linear intercept (Lm),but no change in lung compliance.Furthermore,neither strain had a decrease in lung elastin by immunostaining or by elastin assay methods.In contrast,Cavarra et al.(14)and Takubo et al.(137)found that the pallid mouse,a strain with low levels of ?1-antitrypsin,developed signi?cant air space
enlargement,had a 19%decrease in lung elastin content,and signi?cant increases in compliance (Fig.4),thus more closely resembling human emphysema.Similar results have been found by us in the AKR/J mouse strain (36)and in the guinea pig (18,155).
Which of these approaches is “correct”?There is no simple answer to this question;at the present time,there are no de?nitive studies that illustrate a direct correlation of air space enlargement and compliance or a level of air space enlarge-ment beyond which alterations of compliance would be ex-pected,and it is unclear whether more marked emphysema (as de?ned in this way)is a better approximation to human disease mechanisms than less marked emphysema.Furthermore,the marked laboratory-to-laboratory variations in response in even the same (notional)mouse strains make comparisons almost impossible.
As a general rule,the greater the severity of emphysema,the greater the chance that a protective intervention is really signi?cant,rather than just a minor experiment-to-experiment variation,but this idea must be tempered by the realization that none of these models translates to human disease in any direct fashion,and an intervention that ameliorates fairly mild em-physema might turn out to be mechanistically important.For example,there is increasing evidence for the importance of
immunological reactions in the genesis of emphysema (1).In their investigations relating to the role of CD4and CD8T-lymphocytes in cigarette smoke-induced emphysema,Maeno et al.(81)found
a 19–21%increase in air space size in wild-type C57/Bl/6J
mice with a respective increase of 19%and 0.4%in CD4?/?
and CD8?/?cell-de?cient mice.One must ask,therefore,
whether the lack of lung mechanics invalidates the importance
of this mechanistic data.
Normally lung mechanics in mice and other rodents are
performed at the end of the experiment with the animal alive,
anesthetized,paralyzed,and with the chest opened or closed.
Mechanics measurements done in dead animals are not as
accurate or reproducible,and measurements deteriorate as
times from death increase.Lung mechanics and other pulmo-
nary function tests can be easily measured in guinea pigs (155)
or other large rodents utilizing a body plethysmograph,a
tracheal canula,and an esophageal tube to obtain P-V curves,
FRC,and other lung volumes.Flows can also be measured by
rapidly de?ating the lungs from TLC applying an airway
pressure of ?50cmH 2O.Measurement of lung mechanics can
also be performed in mice,although it is technically more
dif?cult,and we and others have used a small animal ventila-
tor,Flexivent Scireq (Montreal,Canada),equipped with the
necessary hardware and software to accurately measure elas-
tance,resistance,and tissue resistance along with inspiratory
and expiratory P-V curves.
Lung mechanics can also be measured in live rodents,
including mice,with the expectation of recovery after the
measurements (nonterminal event).After anesthesia,the ani-
mals can be (relatively)easily intubated and subsequent mea-
surements obtained (11,140).The obvious advantage of this
technique is that it allows the documentation of functional changes over time in the same animal (161).
Vendors
There are large numbers of animal suppliers available in North America,Europe,and Asia;regular strains,transgenics,genetic models,etc.,are available through these suppliers.The web site https://www.doczj.com/doc/ed6510579.html, is a useful and informative
starting
Fig.4.A :partial pressure-volume (P-V)curves between 3and 9cmH 2O pressure in the NZW Lac/J (resistant strain),C57BL6/J (moderately suscep-tible strain),and AKR/J (super-susceptible strain).[From Takubo et al.(137).]
Pallid P-V curves have been added to illustrate the susceptible strain from our
prior publication.SJ/L and A/J curves were superimposable to the C57BL/6/J P-V curves.*P ?0.001;**P ?0.0001.B :P-V curves in an emphysematous
mouse (A )and a control (B ).Curves were done in the traditional way in?ating the lung to 30cmH 2O and measuring pressure at the airway opening.The maneuver was done both in step-wise,no ?ow,and slow de?ation methods.Both curves are superimposable.The Flexivent machine was used.Invited Review
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point where most of the best suppliers of mice are listed with their www portals.It should be kept in mind that at least for the “regular”nonmodi?ed strains of mice,differences in response to speci?c manipulations in the same strain from different suppliers may occur.In one of our laboratories (M.Cosio),we have identi?ed changes in response to a manipulation (cigarette smoke)in the same strain from the same provider,after a colony had to be restarted from frozen embryos.Cigarette Smoke Exposure Model of COPD Given its position as the predominant cause of COPD,animal models using smoke exposure would appear to be the logical choice for investigation,but it is only relatively recently that such models have been created.We have recently re-viewed the ?ndings and conclusions from smoke-induced an-imal models of COPD (16);here we will consider only the advantages and limitations of such models.Methods of smoke exposure.There are a variety of commer-cial and home-built smoking machines that have been used with animal models;some systems use nose-only exposures and some whole body exposures.There is a theoretic negative to whole body exposure in that the animals may ingest nicotine or tar substances when cleaning their fur.However,Mauderly et al.(92)showed that although this was a factor,there was less carboxyhemoglobin in the animals given whole body exposure,and these animals had less weight loss than did animals given a nose-only continuous or intermittent types of exposure.When establishing any smoke exposure model,measure-ment of either serum cotinine (a nicotine metabolite)or blood carboxyhemoglobin (COHb)is useful as a method of con?rm-ing the relative amount of smoke exposure.In our guinea pig nose-only exposure model,we ?nd an acute post-smoke car-boxyhemoglobin of 15–20%and a chronic blood carboxyhe-moglobin level of ?5%,the latter comparable to that seen in many human smokers.In humans,the dissociation of carboxy-hemoglobin when breathing air results in a half-life of ?4–6h;animal data are not available,although COHb levels rapidly decline after smoke cessation (43).A serum cotinine level of 200–250ng/ml is equivalent to a mild to moderate human cigarette smoker (31,43,103,127).Choice and limitation of species.All modern studies of cigarette smoke-induced COPD have used small laboratory animals including mice,rats,and guinea pigs.Guinea pigs offer numerous advantages,including easily recognizable em-physema,more marked small airway remodeling than mice,and the development of considerable goblet cell metaplasia in the airways.Mice and rats develop only minimal goblet cell metaplasia in the smaller airways,although rats do demonstrate metaplasia in the larger airways.As well,it is easy to perform extensive pulmonary function evaluations along with cardiac catheterization in guinea pigs (155,162).Their main disadvan-tages are cost,the lack of tools for performing molecular studies,and the limited number of commercial antibodies that cross react with guinea pig proteins.In our experience,rats are a poor model for COPD.In most investigators’hands,they develop minimal disease,or,if smoked too heavily,develop nonspeci?c particle overload effects (see,for example,Ref.132).Most recent models of COPD have used mice.Detailed pulmonary function is technically dif?cult,although possible,in mice (36,84,137),but P-V curves and compliance changes can be measured fairly easily.Mice offer the advantages of low cost,extensive gene/protein sequence/antibody availability,and,most important,the availability of numerous naturally occurring mouse strains with different reactions to smoke and the ability to produce animals with genetic modi?cations that shed light on speci?c processes within COPD.
One point that confounds all of the literature on cigarette
smoke-induced models is laboratory-to-laboratory variations in
response.Even when the same mouse strain is used,different laboratories get quite different degrees of disease.For example,Churg et al.found increases in mean air space size (Lm)of 37%(20)and 38%(19)after 6mo of exposure using C57Bl/6mice,but Cavarra et al.(14)and Guerassimov et al.(36)reported increases of only 14%and 13%in the same strain.In part,this problem is caused by differences in the strains from different providers,technical differences in morphometric techniques,the use of different smoking systems (nose only vs.whole body,smoke concentration,smoke time,and frequency),and by differences in the types of cigarettes smoked.It is our impression that currently available cigarettes (such as Ken-tucky 2R4F or 2R5F)cause considerably less emphysema than older ones (such as Kentucky 1R1or 2R1),and this is a serious problem,both in terms of producing disease and particularly in terms of examining interventions,since one never knows whether interventions that ameliorate small increases in air space size would really be effective with more severe disease.Cigarette smoke-induced models of emphysema.Emphy-sema is the anatomic lesion that,historically,has attracted the most attention in patients with COPD,and,similarly,has been the
focus of most animal models (Figs.5,6).Except for ?1-antitrypsin-de?cient pallid mice (14,137),models of smoke-induced emphysema typically produce dilated alveolar ducts;these changes progress with increasing amount of smok-ing and are,anatomically,similar to a mild form of the centrilobular emphysema commonly found in human cigarette smokers,but the overt tissue destruction seen in humans is much harder to demonstrate in animals (14).The parenchyma between the dilated alveolar ducts is abnormal,with increases in size and number of the pores of Kohn that connect adjacent alveoli (154),a phenomenon also found in the lungs of human cigarette smokers (21,65,101).There is evidence that women are more susceptible to the effects of smoke than men (29,70,85,148)and female A/J mice develop emphysema earlier than male A/J mice (84).
The lesions produced in laboratory animals can be subtle,even on microscopic examination;thus morphometric analysis is required to assess the degree of damage.Measurements of air space size (Lm)or surface-to-volume ratio (Sv)(reviewed in Ref.141)and sometimes destructive index (14)are used for
this purpose.Arguably,it would be better to measure only alveolar ducts,since that is the major site of the abnormality,and such measurements do provide more marked differences between smoke-exposed and control animals (see Ref.19,for
example).However,measurements of alveolar duct air require greater morphometric effort (137)and are not widely used.Smoke-exposed guinea pigs and mice show physiological changes that mimic mild COPD in humans (18,36,84,137,155).The residual volume increases as does functional residual capacity and total lung capacity.The P-V curve is shifted upwards and to the left,compliance increases,and the ?ow
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volume curve demonstrates diminished ?ow at the lower lung volumes.Ratios of FEV/FVC also decrease.These abnormal-ities progress with increasing length of smoke exposure (155).
Cigarette smoke-induced small airway remodeling.Small
airway remodeling (increased matrix components,in?amma-
tory cells,and goblet cell metaplasia in the airway wall with
luminal narrowing,distortion,and obstruction by mucus)is
now accepted as an important cause of air?ow obstruction in
human smokers (46,47,98,107,156).These changes increase
with increasing Global Initiative on Chronic Obstructive Lung
Disease (GOLD)stage (47).Excess mucus secretion is also
thought to be important in the pathogenesis of acute exacerba-
tions of COPD (reviewed in Ref.6).
In animals,early smoke inhalation studies demonstrated
increases in the numbers of goblet cells in the proximal trachea
in a variety of species (7,42,57,69,108,117),but there is a
considerable species effect and a large degree of variation in
the degree of metaplastic change.In the guinea pig model,
chronic smoke exposure induces secretory cell metaplasia in
the small airways (158),and similar to humans,smoking
cessation reduces the degree of metaplasia (160).
Little attention has been paid to small airway wall remod-
eling in animal models,probably because these changes are too
subtle to be reliably picked up by casual microscopic observa-
tion,and some deny their existence (84).Nonetheless,mor-
phometric analysis con?rms that the small airway walls are
thickened in both mice and guinea pigs after long-term smoke
exposure (10,17,18,159).Most of the increased wall area is
composed of collagen and ?bronectin,and smooth muscle is
not increased (10,17,95,159).There is also increased collagen
?ber alignment and more dense packing of the ?bers,?ndings
that imply increased mechanical stiffness in the airway wall
(159).In guinea pigs,the severity of changes in collagen
structure correlates negatively with PEF and FEV 0.1/FVC,and
positively with airway resistance.Physiological lung volumes
correlate with air space size,but RV and TLC also are related
to airway wall thickness (159).
Small numbers of lymphocytes are found around the bron-chioles in mice and guinea pigs and are analogous to the lymphoid aggregates seen in humans.Smoke causes quite marked increases in the number and size of these aggregates (9,
10,146).
Cigarette smoke-induced vascular remodeling and pulmo-
nary hypertension.In subjects with established pulmonary
hypertension related to COPD,there is intimal thickening and
muscular hyperplasia in the normally muscularized vessels
adjacent to the bronchioles and increased muscularization of
the usually partially muscularized arteries adjacent to the
alveolar ducts (reviewed in Refs.3,157).
In Hartley strain guinea pigs,chronic cigarette smoke expo-
sure produces about a 25%increase in pulmonary arterial
pressure (160,162),which,interestingly,appears after 1mo of
smoke exposure and does not progress.Similar to human
disease,PHT in guinea pigs is associated with muscularization
of the small,normally poorly muscularized pulmonary arteries
adjacent to the alveolar ducts (162).Increased pulmonary
artery pressure (?30%)has also been reported in one study of smoke-exposed rats (72)but thus far has not been reported in mice.Vascular remodeling,however,has been reported in
mice,and there are apparent strain differences in the severity of
remodeling (100).
Conclusions regarding smoke-induced models.Cigarette smoke exposure to animals appears to be the best
approxima-Fig. 5.Example of centriacinar emphysema in a human.Note that the respiratory bronchiole and alveolar duct complex is distorted and destroyed.Aggregates of pigmented alveolar macrophages can be found (arrow).Bar ?1,000?m;H&E
stain.Fig.6.Example of a guinea pig emphysema model induced by exposure to cigarette smoke for 6mo.A shows a representative control lung,whereas B shows a representative smoke-exposed lung with an overall increase in air
space size of ?43%.Bar ?200?m;H&E stain.
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tion to human COPD:a form of emphysema,small airway remodeling,and pulmonary hypertension can be produced (Table 1).Mucus hypersecretion is more problematic.One important downside to these models is that production of either emphysema or small airway remodeling takes months of smoke exposure (usually on the order of 6mo)and thus is an expensive and time-consuming proposition.However,the most important limitation of smoke models,no matter what species is selected and no matter how long animals are smoked,is that the disease they produce is mild,probably equivalent to human GOLD (24,115)stage 1or 2disease.In humans,the majority of morbidity and mortality occurs in patients with GOLD stage 3or 4disease (FEV 1?50%predicted),but the type of severe emphysema or severe small airway remodeling found in GOLD stage 3or 4patients simply cannot be reproduced in
animals using smoke https://www.doczj.com/doc/ed6510579.html,st,in advanced GOLD stage patients,COPD frequently progresses even after the patient stops smoking,but this phenomenon has not been reproduced in animal models,where cessation of smoke exposure leaves the animals with stable,nonprogressive emphysema,and gob-let cell metasplasia regresses (160).Apoptosis as a Model of COPD There are several excellent recent reviews of this subject (144,145,151,163).The idea that emphysema occurs as a failure of the lung maintenance and repair system follows from studies that have induced apoptosis by interference with vas-cular endothelial growth factor (VEGF)(60)or its receptor (VEGF-R)(139)or the VEGF-VEGFR complex (88).Apop-tosis generally involves activation of the caspase cascade and can be induced by intratracheal instillation of activated caspase-3as well as other agents (2).Activation of ceramide,an upstream regulator,(111)will also induce apoptosis.These models induce air space enlargement in a very short period of time (Fig.7)but do not appear to affect the airways.Inhibition of VEGF receptors suf?cient to induce air space enlargement also causes pruning of the vascular bed and pulmonary hypertension (60,138,143,150),with increased
muscularization of the small pulmonary arteries.Conclusions.Although inhibition of the VEGF system ap-pears to result in relatively permanent air space enlargement,instillation of a single dose of activated caspase-3produced
initial air space enlargement followed by diminution of air
space size within 15days (2).Thus far,none of these models
has been shown to be associated with matrix breakdown,an event that is believed to be central to the pathogenesis of emphysema (reviewed in Ref.16),nor have they been shown to reproduce the physiological changes of emphysema.Finally,it is important to recognize that apoptosis has only been found to be increased in human lungs with severe emphysema,and analysis of the smoke-exposed mouse or guinea model has shown inconsistent results,with no signi?cant increases in the number of apoptotic cells in some studies (Churg A and Wright JL,unpublished data)(25),whereas one study found focal areas of apoptosis,but only in DBA/2and not in C57Bl/6mice (5)(Table 2).
Elastase Emphysema Model
The current generally accepted hypothesis of cigarette smoke-induced emphysema is the protease-antiprotease hy-pothesis.This hypothesis developed out of two key observa-tions:?rst,Laurell and Eriksson (71)reported that patients who were de?cient in ?1-antitrypsin developed early emphysema.Second,in 1965,Gross et al.(34)described the induction of
emphysema in rats by instillation of the plant protease,papain.
These observations led to the idea that emphysema develops as
a result of a cigarette smoke-mediated in?ux of in?ammatory
cells into the lower respiratory tract with resulting release of proteases that destroy the parenchymal matrix.
The protease-antiprotease hypothesis was in large measure formulated from experiments in which emphysema was caused by administration of elastolytic enzymes,either by intratra-cheal instillation or aerosol inhalation;these models are cur-rently referred to by the generic term “elastase emphysema”
Table 1.Pros and cons of cigarette smoke-induced model of COPD Pros:Induced by same insult as in humans Produces emphysema,airway remodeling,and vascular
remodeling/pulmonary hypertension in selected species Produces physiological alterations similar to humans
Cons:Does not produce the severe disabling disease seen in humans Requires several months of exposure Lesions do not appear to progress after cessation of smoke
exposure Fig.7.Example of a mouse apoptosis em-
physema model induced by installation of
active caspase-3.A shows a representative
control lung.B shows a representative image
from lungs 48h after installation of active
caspase-3,with an overall increase in air
space size of ?4%.The arrowheads indicate
enlarged air spaces.Bar ?50?m;H&E
stain.[From Petrache et al.(110).]
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(Fig.8).A variety of enzymes have been used for this purpose (52,53,74,75),and one of the key ?ndings in this regard was that only enzymes that could degrade intact elastin-produced emphysema;collagenases by themselves were ineffective (75,136).These ?ndings led to the current belief that destruction of the elastin framework of the lung is the fundamental patholog-ical abnormality in emphysema.While papain was used in initial studies,its elastase activity depends on its purity and source (75).Almost all recent studies have used either porcine pancreatic elastase (PPE)or human neutrophil elastase (HNE).PPE offers the advantages of being inexpensive and easy to obtain.Consistent quality human neutrophil elastase is now available but considerably more expensive.Experiments using crude preparations of human neutrophils ?rst and then puri?ed HNE were in fact instrumen-tal in arriving at the notion that smoke-evoked in?ammatory cells release elastolytic proteases that can destroy the lung (52,53,75).From a practical point of view,it is important to note that PPE and HNE do not have the same proteolytic spectrum;they have different primary endogenous inhibitors [?1-anti-trypsin for HNE,but ?2-macroglobulin for PPE,although
?1-antitrypsin is also effective (133)],and,even more impor-tant,a particular exogenous inhibitor may not inhibit both of these enzymes (for example,see Refs.67and 68and see below).This is a crucial point to bear in mind if one is attempting to show ef?cacy of a novel inhibitor.A wide variety of animals have been used in the elastase emphysema model (52,75).Much of the early work,and some recent work,was done in hamsters,based initially on the empiric observation that hamsters appeared to develop more severe dis-ease.The reason for this turns out to be that hamsters normally have relatively low levels of ?1-antitrypsin (52,75).Con-versely,
several investigators showed that administration of ?1-antitrypsin or normal serum but not serum from persons de?cient in ?1-antitrypsin prevented or ameliorated elastase
emphysema (58,86);this observation again was important in
formulating the protease-antiprotease hypothesis.
Table 3lists pros and cons of the elastase emphysema model.The major advantages are that emphysema can be rapidly induced by a single treatment with an inexpensive
reagent,which makes it vastly cheaper and easy to run than a 6-mo smoke exposure.Disease severity can be controlled by
selection of enzyme dose.As opposed to smoke-induced em-
physema,it is relatively easy to produce severe emphysema
with elastase instillation,and this facilitates detection of pul-monary function abnormalities,particularly when evaluating the effects of an inhibitor or intervention such as lung volume reduction surgery or abnormalities in connective tissue elastic-ity (15,51).Biochemical and in situ hybridization studies have shown that over time there is increased synthesis of elastin and to a lesser extent collagen (77),making the model a candidate for studies on alveolar repair and regeneration [however,stud-ies using all trans retinoic acid and/or bone marrow cell instillation or various colony-stimulating factors to attempt to drive repair are contradictory (49,50,76,102,164)].But probably the most important current reason to use the elastase emphysema model is the situation where an intervention re-quires scarce/expensive resources that would not be feasible for a 6-mo smoke exposure.For example,Houghton et al.(48)recently showed that antibodies against elastin fragments ame-liorated the in?ammatory response and emphysema in mice given PPE.
The major disadvantages of the elastase emphysema model center around two related problems:1)although both elastase and cigarette smoke produce emphysema through proteolytic attack on the lung matrix,the detailed mechanisms of this process are probably very different;and 2)the mechanisms behind elastase emphysema are not clear.Both light micro-scopic and electron microscopic studies (52,66,165)show rapid loss of elastin after enzyme instillation,but the disease progresses (depending somewhat on animal and dose)long after elastase activity can no longer be detected.In fact,the measured half-life of HNE or PPE in this model is ?45–50min (133).It has been suggested that progression is caused by enzyme bound to the matrix and not detectable with conven-tional assays,but a late intervention (day 20)with an
elastase Fig.8.An example of a murine emphysema model induced by elastase.The ?gure contrasts a representative control lung to a representative elastase-treated lung,with an overall increase in air space size of ?60%;H&E stain.Photos courtesy of Dr.S.Shapiro,University of Pittsburgh,PA.
Table 2.Pros and cons of apoptosis model of COPD Pros:Short-term model Induces enlarged air spaces (?“emphysema”)and vascular remodeling
Cons:“Emphysema”not permanent Apoptosis not consistently present in animal models of cigarette smoke-
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inhibitor did not prevent progression (79),which the authors interpreted to mean that other processes are involved.Single dose elastase inhibitors appear to work when given immedi-ately before or immediately after elastase administration,but are not effective if given 4or 8h later (35,63,134).However,two synthetic serine elastase inhibitors [ICI 200,800and ICI 200,855(153)]prevented both PPE and HNE driven emphy-sema when started 24h after enzyme dosing and continued on a daily basis.Other novel elastase inhibitors/elastin protectors (13,67,80)also provide some degree of protection.In fact,the model is certainly more complicated than pure elastase attack.Elastase emphysema itself is subtly different from human emphysema,with pure alveolar enlargement pre-dominating in elastase emphysema and increased alveolar wall destruction (measured by the destructive index)in human (23)and to some extent in murine smoke-induced emphysema (84).Soon after enzyme administration,there is both hemorrhage and an in?ammatory exudate which includes neutrophils and macrophages in the lower respiratory tract,and there is up-regulation of a variety of proin?ammatory mediators such as TNF ?,IL-1?,IL-6,and IL-8(28).It has been claimed that there are increased numbers of apoptotic parenchymal cells (126,164),but these studies have only used TUNEL staining and hence are inconclusive.In?ammatory processes clearly play an important role since mice lacking both TNF ?and IL-1receptors are ?80%protected against PPE-driven emphysema and in?ammation (78),as are mice overexpressing CuZnSOD (26)and mice given the anti-in?ammatory agent HO-1by
adenoviral transfer (129).Elgin-C,which inhibits rat NE but not PPE,was protective against PPE-driven emphysema,sup-
porting a role for endogenous elastases (68).On the other hand,a broad spectrum MMP inhibitor decreased emphysema but not in?ammation (8).One further practical complication is that animal age appears to be important,but the literature is contradictory,with claims that either younger or older animals are more resistant to PPE (33,59,87).Conclusions regard the elastase emphysema model.Overall,these ?ndings indicate that although elastase emphysema is simple to produce,the model is not mechanistically simple at all.At best,elastase emphysema is a screening technique for mechanisms and/or interventions that could apply to human smokers;however,this approach is fairly far removed from smoke exposure and in our opinion is generally inferior to animal models using smoke exposure.
Starvation-Induced Model of COPD
In humans,chronic malnourishment or actual starvation appears to be associated with emphysema-like alterations of the lung parenchyma (reviewed in Ref.22).Starvation
models in rats have varied in the amount of reduced feeding
and also have varying degrees of alteration in lung structure and physiological changes (119,121–125).Alterations of
lung mechanics were generally characterized by a decrease in lung volume related to transpulmonary pressures,and in one study (125),the shift in the saline P-V curve was accompanied by signi?cant reductions in elastic recoil,and chord compliance (slope of the straight line between 80and
40%maximal lung volume)was increased by 47%,with a
53%increase in air space size.Mechanical changes reverted
to normal with refeeding (123),and although bulk lung hydroxyproline also reverted to normal,elastin remained decreased (122).Cycles of starvation and refeeding resulted in overall ?ndings similar to those above,but there was also a decreased alveolar surface area interpreted as a decreased number of alveoli (120).Calorie depletion accompanied by protein depletion showed the unexpected ?nding of less severe emphysema than in those animals with similar caloric intake that included protein.All of the calorie-restricted groups had smaller lung volumes and fewer alveoli than did the control group indicating that the effect of starvation is a lack of lung growth.
A comparison between starvation (45%loss of body wt)and elastase-induced emphysema showed considerable differences between the two models (41).While elastase-induced emphy-sema mimicked human emphysema,with increased lung vol-umes and subdivisions of lung volume,a shift to the right of the P-V curve,and reduced expiratory ?ow rates,the P-V curve in the starvation group was shifted to the right,the lung volumes were not increased,and there was no decrease in air?ow.Both groups had enlarged air spaces suggesting that the two models may represent lung destruction in the case of elastase emphysema and alterations of lung growth in the case of starvation emphysema.But recent studies in mice (89,91)have questioned this explanation since alveoli appear to be lost early in calorie restriction and gained after refeeding.Starva-tion appears to stimulate caspase gene expression and Fas
Table 3.Pros and cons of elastase model of COPD Pros:Single treatment (as opposed to 6mo of smoke)/economic and easy Rapid onset Can produce severe disease depending on dose Progressive in some hands Easy to measure functional changes Good choice when dealing with scarce resources where impossible or too
expensive to run a 6-mo smoke Possibly relevant to certain aspects of regeneration of elastin network
Cons:Choice of enzyme,animal species,age of animals,dose protocol crucial for
consistent results Neutrophil elastase and porcine pancreatic elastase appear to act somewhat differently Relevance to mechanisms that actually occur with smoke exposure is unclear Elastase emphysema does not model all the very complicated events seen with smoke Relevance of therapies/interventions even less clear than with animal models of smoke exposure Table 4.Pros and cons of starvation model of COPD Pros:
Short-term model with relatively consistent results
Cons:
Does not match human physiological changes
May represent a model of abnormal lung maintenance/repair/abnormal lung growth
Ethical considerations of induction of 45%weight loss by starvation
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death receptor expression,with depression of cell replication,while refeeding increases cell replication,DNA synthesis,and
increased RNA expression for a large number of cell replica-tion-related genes.Conclusions regarding starvation models.Starvation models are relatively short term in duration and produce enlarged air spaces,but they do not produce the physiological changes of emphysema and more likely represent models of abnormal lung growth or abnormal repair (Table 4).No airway or vascular alterations have been described.LPS-Induced Models of COPD
LPS is a glycolipid component of gram-negative bacterial
cell walls and is used because of its proin?ammatory effects.It is present as a contaminant in cigarette smoke,air pollution,
and organic dusts.In humans,chronic exposure to LPS-laden
dusts results in decreased lung function (reviewed in Ref.147).
In the acute model,LPS induces a mixed in?ammatory reac-
tion with increases in neutrophils (8)and increases in BAL
TNF ?and IL-1?,and increases in MMP-9and MMP-12(73,
147)(Fig.9A ).In mice (147),repeated intratracheal LPS
installations induces marked in?ammation with predominately
CD4?T cell lymphoid aggregates around the airways and
large vessels,and an increase in intra-epithelial bronchiolar
epithelial cell lymphocytes.This was associated with large
numbers of intra-alveolar macrophages.Cytokine mRNA pro-
?les showed increases in TNF ?,IFN ?,and IL-18,a ?nding
that persisted after exposures ceased.
With chronic administration over several weeks,LPS instil-
lation produces enlarged air spaces (Fig.9B )and remodeled
airways with thickened walls and increased goblet cells in the
larger airways in mice,guinea pigs,and rats (40,142,147).In
the guinea pig model (40),progressive exposures resulted in a
decline in speci?c airway conductance and an increased dura-
tion of bronchoconstriction after exposure.These changes
resemble human emphysema and small airway remodeling
(albeit again relatively mild)and may be driven by mecha-
nisms that are somewhat similar to human disease;however,
these mechanisms are poorly de?ned.
Conclusions regarding LPS models.LPS instillation is a
short term model which produces some anatomic features of
human COPD,but the in?ammatory in?ltrate differs from that
in smoke models,suggesting different mechanisms (Table 5).
Probably the most important use of LPS is to mimic acute
exacerbations,either given alone or given to animals also
receiving cigarette smoke.The topic of models of acute exac-
erbation has been recently reviewed in Refs.30,62,83.
Genetic and Genetically Modi?ed Models of COPD
Much of the recent literature on COPD models has used
either naturally occurring mouse strains or laboratory-produced
animals that either overexpress or lack particular genes.Sev-
eral excellent reviews of such models are available (12,82,
128,144,149),and we will only comment brie?y here on their
advantages and disadvantages.
Different naturally occurring mouse strains show markedly different responses to smoke which depend in part on naturally occurring levels of antioxidant and antiprotease protection and
in part on genetic factors that control the innate and acquired in?ammatory response to smoke (14,81).Different strains have a variety of genetic differences,and thus one usually cannot attribute different responses to an insult such as
smoke
Fig.9.A :example of a mouse subacute LPS model induced by exposure to LPS for 1h per day for 5days.Note the in?ammatory reaction in the air spaces and interstitium.H&E stain.Magni?cation,??10,courtesy of Dr.M.K.Lee,
Battelle Science and Technology,Richland,WA.B :example of a hamster emphysema model induced by installation of LPS twice per week for 4wk and death after 28days after the ?nal installation.Emphysematous areas are surrounded by relatively normal areas with an overall increase in air space size of ?30%.The arrows point to iron depositions.[From Rudolphus et al.(118).]Table 5.Pros and cons of LPS model of COPD
Pros:
Short-term model with airway and parenchymal changes,and in?ammatory cytokine upregulation
May be helpful in investigations of exacerbation
Cons:
In?ammatory in?ltrate not analogous to that found with cigarette smoke Unclear whether mechanisms involved recapitulate those of cigarette smoke-induced lesions
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to a single gene change,as is possible with man-made genet-ically targeted animals (there are exceptions to this rule,however,such as pallid mice that are ?1-antitrypsin de?cient)(14,137).Genetically altered animals allow investigation of the effects of a speci?c gene/protein on all of the different anatomic
lesions of COPD and potentially also are useful in designing therapeutic agents.Most attention has been directed to models of emphysema,with much less information available about small airway remodeling (64).All such models need to be interpreted very cautiously,because constitutive overexpres-
sion or knockout of a particular gene can interfere with lung development and lung growth,resulting in enlarged air spaces that super?cially appear to represent emphysema but are really developmental abnormalities.Conversely,constitu-tive expression of a gene might also lead to accelerated aging,and accelerated senescence mice are known to de-velop emphysema (130).Several different approaches have been taken to try and get around the issue of developmental abnormalities.Some con-stitutive transgenic models of emphysema have concentrated on selection of strains in which transgene expression is present at a very low level and disease only appears to manifest late in adulthood (27).Such models provide support for the impor-tance of the gene in question but by de?nition cannot rule out either subtle developmental or aging abnormalities.Inducible models generally use constructs in which feeding of doxycycline drives the gene in question with a lung-speci?c promoter.This approach in theory prevents problems with developmental or premature aging effects.However,there may be small amounts of “leakage”of the transgene even in the absence of doxycycline,and it has been reported that the reverse tetracycline-transactivator gene that is used to create inducible constructs can itself can produce enlarged air spaces (130).As well,doxycycline is a matrix metalloprotease inhib-itor that inhibits MMPs such as MMP-2and MMP-9that are thought to play a role in COPD (37).Thus careful controls are essential in interpreting such https://www.doczj.com/doc/ed6510579.html,st,it is important to remember that overexpression of a particular gene may lead to production of huge amounts of protein,and there is no guarantee that “pharmacological”doses of a particular protein act in a similar fashion to physiological doses.Conclusions regarding models using genetically altered an-imals and different naturally occurring strains.These prob-lems notwithstanding,genetically modi?ed animals and natu-rally occurring mouse strains have produced a wealth of data,either by themselves or when such animals are exposed to cigarette smoke.We have recently reviewed the topic of smoke-induced models and the mechanisms behind such mod-els in this journal (16).How closely these models mimic the pathogenesis of human disease is still open to question (4).Conclusions A variety of quite widely differing types of animal models has been created to attempt to reproduce human COPD.None of them succeeds perfectly in this attempt,and all have prob-lems.Nonetheless,cigarette smoke-induced models of either naturally occurring or man-made animal strains appear to be the best choice for most purposes if one has the resources and patience to carry out the very long exposures required.Elastase emphysema has a limited role,particularly when resources are scarce.Models that examine failure of lung maintenance or failure of repair will probably be important in understanding the pathogenesis of emphysema,but as yet don’t clearly approximate events in the human lung.
GRANTS
This work was supported by Canadian Institutes of Health Research Grants 42539and 81409,a grant from the Heart and Stroke Foundation of British Columbia and Yukon,and the Lloyd Carr-Harris Foundation.
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最新人教版新起点四年级英语上册期末测试卷 班级_________ 姓名__________ 成绩________ 一.写出26个字母的大小写。 二.单项选择。 ( )1.I’m going to make ___invitation. A.a B.an C./ ( )2.I have ____. A.scisors B. scissors C. scissor ( )3.Happy ____day. A.Teacher B. T eacher’s C. Teache rs’ ( )4.Do you have glue? ______. A.Yes. I do. B.No. I do. C.Yes.I don’t. ( )5.Is it time for lunch?_______. A.Yes. it is. B. No. it is. C. Y es. it isn’t. ( )6.Do you have a pen? _______. A.Y es. I don’t. B. No. I do. C. Yes. I do. ( )7.Can I have more turkey?_______. A.Yes，you can . B.No ，you can’t. C.Y es, you can’t. ( )8.She often ____cycling. A.play B.go C.goes ( )9.He often ___tennis. A.go B.goes C.plays ( )10.Lily____hockey. A.are good B.is good at C.are good at ( )11.Do you often play tennis?_____. A.Yes, I am. B.Yes, I do. C.N o, I’m not. ( )12.Are you good at it? ______. A.Yes, I am. B.N o, I don’t. C.Yes, I do. ( )13.T they can’t play ___the field. A.in B.on C.at ( )14.They can throw it ___player___player. A.from···to··· B.from···for··· C.for···to··· ( )15.I have some ___. A.boxs B.boxes C.box ( )16.Today basketball need 2teams of ___player. A.6 B.7 C.5 ( )17.___do you do for fun? A.How B.W hat’s C.What ( )18.One student ___to do puzzles. A.like B.does like C.likes ( )19.Do you like to go to concerts? Yes,____. A.I do. B.I like. C.I can. ( )20.Is here your homepage? Yes, ___. A.it is. B.here is. C.there is. ( )21.why do you like to drive bumper cars? ____it’s exciting. A./ B.Because C.Do ( )22.I’m___firecrackers. A.light B.lights C.lighting ( )23.___careful! A.You B.I’m C.Be ( )24.My leg____. A.hurt B.is hurt C.hurts ( )25.Call_____ambulance. A.a B.an C.the ( )26.We are taking you ___the hospital. A.for B.go C.to ( )27.Good morning!_____.
Unit 1 My Classmates 单元教学目标 能力目标： 1、能够正确使用描述人物外貌和性格特征的单词及相应的物主代词。 2、能够使用本单元所学的句型，谈论自己的同学或朋友。 3、能够拼写与同学有关的词汇，能仿写与同学有关的句子和短文。 4、能够开展课堂活动，交换与同学有关的信息，进行听、说、读、写技能训练。 5、能够通过故事的学习，提高朗读、理解的能力和积极参与故事表演的能力。 知识目标： 1、能够听、硕、读、写与描述人物有关的17个单词：twins, blond, curly, slim, straight, freckles, talkative, honest, careless, popular, hardworking, shy, lazy, quiet, helpful, friendly, funny以及三个形容词性物主代词：his, her, their. 2、能够理解故事中出现的新词汇：Camp Pine, paddle, trick, laugh, fishing pole, fall. 3、能够听、硕、读、写与同学有关的以下交际用语：“What does your new classmate look like? Her hair is blond and curly. Tell me about your classmates. What’s he/she like? He/she is friendly.” 4、知道字母组合wh在单词中的发音，并根据读音规则正确读出相关的单词和句子。 5、能够在仿写练习中正确携句子，能够正确使用标点符号和书写隔世。 Lesson 1 1--1 教学重点： 1、认读本书中新出现的人物。 2、词汇：his, her, their, twins, blond, curly, slim, straight, freckles. 3、句型：（1）What does your classmate look like? (2) Tell me about your classmates. (3) What’s he/she like? He/she is friendly. 教学难点： 1、三个形容词性物主代词his, her, their的理解。 2、可数名词既不可数名词与be动词的搭配，如：“Her hair is ...Their eyes are (i) glasses are …” 教学准备：录音机、单词卡片、教学挂图 教学过程： 一、教师组织学生就主题情景图进行讨论，如：“Look! This is a classroom. What can you see in the classroom? Who are they? What color is Ann’s hair? 等等。在讨论的过程中，教师通过指挂图、做手势等方式，引导学生初步理解，学习生词及主句型。 二、Do Part A: listen and write down their names. 1.First time listen and find which one is right. 2.Second time listen and write. 3.Third time check and correct the answers. 三、Practice Part B: Choose, ask and answer.
新起点人教版英语四年级上册教案 人教版英语四年级上册备课 执教教师 于涵
教学进度表 四年级英语任课教师：于涵
Unit1 单元分析 【内容来源】人民教育出版社（一起点）四年级上册Unit1 Sports and Games 一、语言技能目标 第一层次：
1. 能够听懂、会说running, basketball, roller skating, jumping rope和ping-pong等描述运动项目的单词和短语；能够根据语境恰当使用它们,并能借助拼读规律认读并识记单词。 2. 能够听懂、会说用来询问某人体育锻炼习惯及特长的功能句：Do you often ...? Are you good at it?及答句：Yes, I do. / No, I don’t.和Yes, I am. / No, I’m not. 能够初步在情境中进行口头运用,并能根据问句写出答句。 3.能够读懂关于朋友体育爱好和锻炼习惯的日记,提取关键信息匹配图片,并能使用结构I often ... I am good at ... 简单地书面介绍自己的体育爱好情况。 4. 能够比较流利地朗读小文段。 5. 能够借助熟悉的旧单词感知字母组合or的发音规律,借助图片理解新单词的词义,并利用该规律 拼读新单词,进而基本能做到“听词能写,见词能读”。 第二层次： 1. 能够借助图片读懂其他国家小学生的体育爱好情况,并模仿范例进行小制作。 2. 学唱与本单元话题相关的歌曲。 3. 能够借助图片和其他阅读策略读懂Story Time的故事,完成故事后的句子排序活动,并在教师的指导下表演小故事。 二、其他目标 1. 通过谈论体育运动增强体育锻炼意识。 2. 能够积极参与课堂上的各种活动或游戏,并能与同伴合作完成活动或游戏。 3. 能够通过看图或提问题预测文段内容,初步养成读前或听前预测的习惯。 4. 能够在活动中认真倾听同伴发言,并积极表达自己的观点。 Unit1 第1课时教学设计 【内容来源】人民教育出版社（一起点）四年级上册Unit1 Sports and Games 【课时】第1课时课件展示 一、教学目标 1. 能够听懂、会说running, basketball, roller skating, jumping rope和ping-pong等描述运动项目的单词和短语；能够根据语境恰当使用它们,并能借助拼读规律认读和识记单词。 2. 能和同学用Do you like ...? I like ... It’s interesting. What about you?等句子询问、交流体育爱好。 3. 能够根据自己的体育爱好,选择本课所学词汇完成句子填空,同时强化良好的英文书写习惯。 二、教学建议 A. Look, listen and repeat.
六年级英语分类复习阅读训练题 班级：_______________ 姓名：_____________ （1） Ben: Hi, Kitty. Kitty: Hi, Ben. Ben: Let’s have a camp trip this Saturday. Kitty: Great! What do we need? Ben: We need some bread and drinks. Kitty: What drink do you like? Ben: I like apple juice. What about you? Kitty: Milk. It’s my favourite drink. Ben: All right. Let’s go to the supermarket now. Kitty: OK. But I haven’t got enough (足够的) money. 快速阅读，用T或F判断下列句子的正误。 ( ) 1. Ben and Kitty want to go for a camp this Sunday. ( ) 2. They only need something to drink. ( ) 3. They want to go to the supermarket to buy these things. ( ) 4. Ben’s favourite drink is milk. ( ) 5. Kitty want to take some apple juice with him. ( ) 6. They have enough money now. （2） A: Hi, Ben! It was National Day holiday last week. What did you do? B: On Monday I watched a film with my family. On Tuesday and Wednesday I was at a camp. I played football with my family. On Thursday I played computer games with my friends. On Friday and Saturday I visited my u ncle and aunt. They’re farmers. On Sunday I stayed at home and watched TV. A: Wow, you were busy. 根据对话，在横线上写出动词词组。 Sunday _____________ Monday _____________
Unit 1 My classmates 一、用适当的人称代词和物主代词填空。（每空一词） (1 ) ________am a bus driver.This is ______bus._________like it very much.. (2 ) Hi,________name is Tom.And you ? (3 ) He is a pupil._________name is Peter._________is tall. (4 ) That is ________school.I like it. (5 ) –Look there,is________Miss White ? –Yes,________is a teacher. (6 ) What are ________over there ? __________are monkeys. (7 ) –Where are the pears from ?--__________are from Shandong. --Are ________sweet ? --Yes,_________think so. (8 ) –What can _________do for you?—I’d like some apples. 二、用适当的指示代词填空。 (1) _______keys are mine. Those are yours. (2) -- What’____________? --It’s an egg. --What’s__________over there ? --It’s an orange.
2016—2017学年度第一学期人教版英语四年级上册备课 执教教师 于涵
教学进度表 四年级英语任课教师：于涵
Unit1 单元分析 【内容来源】人民教育出版社（一起点）四年级上册Unit1 Sports and Games 一、语言技能目标 第一层次： 1. 能够听懂、会说running, basketball, roller skating, jumping rope和ping-pong等描述运动项目的单词和短语；能够根据语境恰当使用它们，并能借助拼读规律认读并识记单词。 2. 能够听懂、会说用来询问某人体育锻炼习惯及特长的功能句：Do you often ...? Are you good at it?及答句：Yes, I do. / No, I don’t.和Yes, I am. / No, I’m not. 能够初步在情境中进行口头运用，并能根据问句写出答句。 3.能够读懂关于朋友体育爱好和锻炼习惯的日记，提取关键信息匹配图 片，并能使用结构I often ... I am good at ... 简单地书面介绍自己的体育爱好情况。 4. 能够比较流利地朗读小文段。 5. 能够借助熟悉的旧单词感知字母组合or的发音规律，借助图片理解新单词的词义，并利用该规律拼读新单词，进而基本能做到“听词能写，见词能读”。 第二层次：
1. 能够借助图片读懂其他国家小学生的体育爱好情况，并模仿范例进行小制作。 2. 学唱与本单元话题相关的歌曲。 3. 能够借助图片和其他阅读策略读懂Story Time的故事，完成故事后的句子排序活动，并在教师的指导下表演小故事。 二、其他目标 1. 通过谈论体育运动增强体育锻炼意识。 2. 能够积极参与课堂上的各种活动或游戏，并能与同伴合作完成活动或游戏。 3. 能够通过看图或提问题预测文段内容，初步养成读前或听前预测的习惯。 4. 能够在活动中认真倾听同伴发言，并积极表达自己的观点。 Unit1 第1课时教学设计 【内容来源】人民教育出版社（一起点）四年级上册Unit1 Sports and Games 【课时】第1课时课件展示 一、教学目标 1. 能够听懂、会说running, basketball, roller skating, jumping rope和ping-pong等描述运动项目的单词和短语；能够根据语境恰当使用它们，并能借助拼读规律认读和识记单词。 2. 能和同学用Do you like ...? I like ... It’s interesting. What about you?等句子询问、交流体育爱好。 3. 能够根据自己的体育爱好，选择本课所学词汇完成句子填空，同时强化
盐亭华宸国际学校五年级上册英语复习资料第二部分句子 Yaoyao： Bill： Joy：He’s my favourite teacher. S1:What does he teach? Joy: Linda: Binbin: Linda: Binbin: How much is the Saleswoman: Binbin:OK. Andy:Binbin, Binbin:My favourite TV shows are sports shows. Andy: Binbin:I usually watch them on Saturday Evening. Binbin: Yaoyao:Yes,I do. Binbin:What chores do you usually do? Yaoyao: Binbin: Yaoyao:Three times a week. 第三部分作文
范文一 My Friend I have a friend. Her name is Lily. She is polite. She often says hello and thank you to everyone in the family. She is active. She often answers questions in English class. She is helpful too. She often helps her mother clean the room. She is clever. She is good at maths. She has short straight hair. She has big eyes. She wears glasses. We all like her. Do you want to be her friend? 范文二 My Favourite Teacher My favorite teacher is Mrs. Wu. She teaches us English. She is tall and slim. She has long curly hair. She doesn't wear glasses.She is very interesting. She often sing English songs in class. She is very popular too. She often plays ping pong with us. I like her very much. 范文三 My Favourite Animal My favorite animal is the panda. It lives in the b amboo forest. It eats bamboo. It is black and white. I t's very cute, too. It has a big body and two big eyes.It can climb up trees.What’s your favourite animal? 范文四 Shopping Day Today is Sunday.My parents are going to go shopping in a supermarket.My mother wants to buy a dress and a pair of pants.The pink dress is 100 yuan.The pants are 12 yuan.She likes them very much.My father wants to buy a new pencil sharpener for me.He buys the orange one.It’s only 12 yuan. I like it too. 范文五 My Favourite TV Shows My favourite TV shows are nature shows.I think they are great.I can learn about animals from nature shows.I usually watch them on Sunday.What about you? 范文六 My Day for Chores On Saturday I usually help my mother do chores at home. After getting up ， I usually make the bed. Then I sweep the floor and water the plants.In the morning, I often walk the dog. I help my parents make lunch, too. After lunch, I often do the dishes.In the afternoon,I wash clothes with my mother. I enjoy doing chores.
五年级英语上第六单元+复习2知识点练习 一, 单词与词组 1, __________________ 做家务 2, _____________________ 帮助你的父母3, __________________ 在家 4, _____________________ 打扫房间 5, _________________ 铺床 6, _____________________ 倒垃圾 7, _________________ 洗衣服 8, _____________________ 整理书桌 9, _________________ 扫地 10, ______________________ 帮助某人做某事11, _________________ 多久 12, _____________________ 一天一次 13, _________________ 一周两次 14, ____________________ 一个月三次 15, _________________ 和我玩 16, ______________________ 给植物浇水17, _________________ 喜欢做某事 18, ____________________享受做某事19, _________________ 喂我的鱼 20, _____________________给我打电话21, _________________ 当你有空的时候22, ________________ 晚饭前 23, _________________ 晚饭后 24, ______________________宠物猫 25, _________________ 起床 26, _______________________做午饭 27, _________________ 午饭后 28, ______________________洗碗碟 29, _________________ 忙碌的 30, ______________________进入 31, _________________ 去野营 32, more and more helpful_________ 33, _________________ 病人 34, sick kids ______________ 35, _________________ 使人振奋 36, how to cook _____________ 37, _________________ 100年以后 38, write an ad _____________ 39, _________________ 返回家 40, ________________ 寻找 41, _________________ 关心 42, ________________ 玩游戏 43, ____________________________________________ 愿意做某事 44, be the host of ________________________ 45, a news announcer_____________________ 46, an exciting job ________________________ 47, ___________________举办(穿上) 48, a quiz show __________ 49, ___________________ 来我家 50, arrive at _____________ 51, _____________________________ 举办一次生日聚会 52, _________________ 在周日 53, __________________ 打算，计划 53, turn on the TV _________ 54, ___________________ 跟某人说话 55, have a surprise _________ 56, to one’s surprise __________ 二，字母组合的单词 1, ______椅子 2, ______桃子 3, ________午饭 4, ______捉 5, ______比赛 6, ______痒 7, _____购物 8, _____ 9, _______洗涤 10,_________嘴 11, ______轮子 12,_______胡须 13,_______ 父亲 14, ________ 鱼 三, 词型转换 1, chore (复数) _____ 2, out (对应词) _____ 3, make (现在分词)_______ 4, wash (三单) _____ 5, tidy (三单) _______ 6, usually (同类词)__________ 7, time (复数) ____ 8, I’m (完全形式)_____ 9, feed(现在分词)______ 10, me (主格) __ 11, you’re (完全形式)_____ 12, too (同音词) _____ 13, do (单三) ____ 14, Susan (名词所有格) ______________ 15, before (对应词) _________ 16, get (现在分词) __________ 17, dish (复数) ____________ 8, catch (单三) ____________ 19, kids (同义词) __________ 20, can (否定形式) _______或 _____ 21, I’ll (完全形式) _________ 22, do (过去式) ______ 23,one (序数词) ____ 24, two (序数词) __________ 25, three (序数词)____________ 26,I’d(完全形式) ___________ 27, let’s(完全形式) ____________ 28, they’re(完全形式)______ 29, excite(形容词) ____________ 30, that’s (完全形式) ______ 31, they(物主代词) ____________ 32, see(同音词) __________ 33, talk(现在分词) ___________ 34, lady(复数) ____________ 35, he(物主代词) _____________ 36, shop(现在分词) _______ 37, has(原形) _________________ 38, child(复数)____________ 39, surprise(形容词)___________ 40, become(现在分词)_____ 41, present(同义词) ___________ 四, 句子 1, _____________________________________________________________ 你通常在家做什么家务？ 2, ______________________________________________________________ 我通常帮助我的父母倒垃圾。 3, _____________________________________________________________ 你通常在家做家务吗？ 4, ____________________________________________________________ 是的，我通常在家给植物浇水。 5, ___________________________________________________________ 你正在做什么？ 6, __________________________________________________________ 我正在打扫我的房间。 7, __________________________________________________________ 你多久扫一次地? 8, __________________________________________________________ 我一周扫三次地。 五, 句型转换. 1, Do you help your parents do chores at home ? ______________ （肯定回答） _________________ （否定回答） 2, I usually make the bed at home .（主语改为Binbin） _____________ usually __________ the bed at home . 3, Bob usually tidies the desk at schooln . (改为否定句) Bob ____________usually ____________the desk at school . 4, I usually help my mother make liunch .（改为一般疑问句） ________ _________ usually help your mother make lunch ? 5, I walk the dog in the afternoon .( 主语改为he） _____ _________ the dog in the afternoon .
上册UNIT 1 invitation [ ???????????] crayon [ ??????] scissors [ ?????] pen [???] glue [?●◆?] paper [ ?????] party [ ?????] get ready [???] [ ????] draw [????] cut [ ??] stick [????] write [????] first [?????] next [?????] then [???] finally [ ?????●?] with [???] practice [ ???????] 邀请卡 蜡笔 剪刀 钢笔 胶水 纸 聚会 准备好 画 剪，切 粘贴 写，写字首先，第一其次 然后，于是最后用。。。。。。练习
UNIT 2 sport [?????] game [????] jogging [ ??????] hiking hike [????] tennis [ ?????] cycling [ ????●??] hockey [ ????] baseball [ ???????●] also [ ??●??◆] sometimes [ ????????] often [ ??????] 运动 比赛，竞赛 慢跑 徒步旅行 网球，网球运动 自行车赛，骑自行车兜风曲棍球 棒球，棒球运动 也，同样 有时，不时 常常，经常
UNIT 3 concert [ ??????] go to the movies [ ?◆???] lego drive [?????] pumper car [?????????] puzzle [ ???●] weekend [????????] exciting [?????????] cool [ ◆?●] just [?????] so-so [ ??◆??◆] 音乐会，演奏会 去看电影 乐高拼装玩具 开车，驾驶 碰碰车 智力玩具，拼图玩具 周末 激动人心 棒极了，酷的 只是，不过 平常的，不好不坏的
Unit 1 In China Lesson 1 世界上有许多美丽的国家，其中我们伟大的祖国中国他屹立在世界的东方，她是一位美丽善良的母亲。 在这辽阔的国土上有着神奇的景观，山川与河流，还有一群勤劳美丽而善良的民族。那么，就让我们一起更加真切地了解认识我们的祖国吧。 【知识目标】 1. Be able to listen and read the words and dialogues: morning tea, 早茶 soup 汤; the Potala Palace, 布达拉宫；the Summer Palace, 颐和园； the Terracotta Army 兵马俑。能够运用已学功能句操练本课新学词汇。 2. Be able to ask and answer questions. 【能力目标】 对听说读写能力都要有意识地发展锻炼，学生能够掌握重点单词的读写及其对应含义，这也有助于自然地学习英语句型。What did you go on your summer vacation? I went to... with my parents. 能够用英语流利表达自己过去做的事情，学会用一般过去时回答完别人的问题。 【情感目标】 学习英语最大的乐趣就是能够理解运用。多了解英语与汉语不同的地方，也要善于要拓展学习其他方面，多参加课外活动。老师要学着寓教于乐地教学，让英语学习成为孩子们生活中的一种乐趣，沟通和学习的工具。 【教学重点】. 1. The relevant words and dialogues. 2. Be able to do some free talk with the words.
小学五年级期末考试 英语试卷（人教新起点版） 题号ⅠⅡⅢⅣⅤⅥⅦⅧⅨⅩⅪ总分 得分 I.选出每组单词中画线部分读音与其它两个发音不同的选项。 (共5 小题，计5 分) ( ) 1. A. cow B. brown C. yellow ( ) 2. A. doctor B. short C. morning ( ) 3. A. cake B. cap C. ant ( ) 4. A. engineer B. driver C. English ( ) 5. A. cloudy B. mouth C. young Ⅱ. 看图片,写单词。 (共 5 小题，计 10 分) 6.7. 8.9. 10. Ⅲ. 根据首字母提示并联系上下文写出恰当的单词，使句子或对话意思完整。(共 5 小题， 计10 分) 11.—What color do you like? —I like red, but I d like yellow. 12.—is that boy? The small one. —Oh, that’s Bob. 13.—Mom, I want to go out to play. —OK. It’s cold outside. P on your coat, gloves and boots. 14.—What’s your grandfather l , David? —He is short and has short white hair. 15.It’s r today. Don’t forget the umbrella and the raincoat. Ⅳ. 根据句意及上下文选择合适选项，将其标号写在相应的括号内。(共 5 小题，计 5 分) A. play chess B. Good idea C. They are whales D. Which one E. weather report 16.—Mom, we are going hiking tomorrow. What’s the weather like? —Let’s listen to the . 17.—Are they seals? —No. . They’re very kind and friendly to people.
新起点英语四年级上册 LESSON 1 马上就到教师节了你要为这个晚会做些什么呢？ 我要做一张邀请卡。你呢？我也是，噢，我们一起做吧。 好的，你有剪刀吗？是的，我有 LESSON 2 让我们做一张邀请卡。准备好了，准备好了，你准备好纸。 第一；画画，用蜡笔画画。接下来用剪刀剪。 然后用胶水贴。最后，用钢笔写。 好了好了，多漂亮的邀请卡。 LESSON 3 你好，焦，你要为这个晚会做些什么？我们要准备一些歌舞。 马上就要到教师节了，我要做一张邀请卡。我们一起做吧。 用剪刀剪首先，用剪刀剪一些星星。然后，接着，最后，写上 用钢笔写：教师节快乐； 儿童节快乐新年快乐； 生日快乐；圣诞节快乐。 打扫教室，练习歌曲和舞蹈 做一张请柬擦黑板 同学们，马上就要到教师节了，我们要为晚会准备一下了，你要为晚会做些什么呢？ 亲爱的高老师：我们将在教师节那天会准备一个晚会，请下午四点到我们班来，教师节快乐。四年一班。 LESSON 4 亲爱的焦， 在这天晚上，我们通常在家里举办一个晚会，首先，家人在一吃晚餐，然后，我们在院子里聊天，我们一起赏月，最后一起吃月饼和水果。 你呢，瑶瑶。 亲爱的吉姆 在这天，我们通常去天安门广场，首先，我们看升国旗，然后，我们绕着广场，看看花再照一些照片，最后，晚上一起回到家再看看电视。 你呢，比尔。 亲爱的瑶瑶 在这天，我们通常有一个晚会，首先家人围坐在圣诞树下，接着，我们打
开来自圣诞老人的礼物，然后，我们吃火鸡晚餐，最后，我们一起做游戏，唱歌或看电视。 你呢，焦 在这天，我们通常有一个晚会，我们围坐在圣诞树下，接着，我们打开来自圣诞老人的礼物。我们吃火鸡晚餐，最后我们做游戏，唱歌或者在一起看电视。 今天是我的生日，是快乐的日子，请留下一起玩。 LESSON 5 看这些红黄相间的树叶，它们很漂亮。 现在还没有，我们到树林里去散散步吧。 吃晚餐的时间到了吗？我喜欢火鸡和南瓜饼。 现在还没有，火鸡还在做。 奶奶感恩节快乐，爷爷感恩节快乐，吃晚餐的时间到了吗？我喜欢火鸡和南瓜饼。 嗯，闻起来很香，吃晚餐的时间到了吗，我很爱吃火鸡和南瓜饼。 现在还没有，南瓜饼还在做。 嗯，看起来很好，吃晚餐的时间到了吗，我很爱吃火鸡和南瓜饼。 是的，来，桌子这边，已经好了。 感谢你为我们准备这些食物。 嗯，味道好极了，我能多要一些火鸡吗？ 是的，我很高兴你喜欢。 鲍伯，你还要火鸡或是饼吗？ 噢，是的，两个都要。 鲍伯，你怎么了？ 我胃疼，叫医生。 没关系的，只是吃太多的火鸡和饼了。 LESSON 6 用蜡笔画，我得到100分。 我们正准备9月10号教师节晚会。首先，我们要有用蜡笔和剪子制作邀请卡，接着，我们要打扫教室，然后，我们要做一些歌舞练习，最后，我们要举办晚会。 首先我们要打扫教室。 然后我们要举办晚会。 接着我们要做一些邀请卡 最后我们要准备一些歌舞 LESSON 7 你喜欢打棒球吗？
新起点英语五年级上册知识要点总结 Unit 1 一. 语音 字母组合 bl ,pl ,cl, gl, fl , sl 在单词中的发音 二,词组总结 1. talk about 谈论 2. look at 看 3. take sb to sp 带某人到某地 4. so many 如此多 5. make model planes 制做飞机模型 6. like to do 喜欢做某事 7. like doing 8. forget to do 忘记将要做的事 9. forget doing 忘记已做过的事 10. want to do 想要做某事11. introduce … to 介绍 12. let`s help… 让我们来帮助… .. 13. stay with 和… . 呆在一起 14. in English class 在英语课上 15. play football 踢足球 16. every day 每天 17. Thank you to every one in the family 她经常对家里的每个人说谢谢你。
18. in the family 在家里 19. clean the car 清洁汽车 20. walk the dog 溜狗 21. play with 和… .. 玩 22. become good friends 成为好朋友 23. go to school 上学24. new friends 新朋友 25. birthday party 生日聚会 26. big hands 大手 27. long arms 长胳膊 28. answer the questions 回答问题 29. be good at 擅长 30. be famous for 以… . 而著名 31. play tricks on 和… 开玩笑 32. come to 来到 33. Beijing duck 北京烤鸭 34. at first 起初 35. a lot 许多 36. at camp 露营 37. in a boat 在小船上 38. catch fish 捕鱼 39. put … on 把… 放在… 上 40. pick up 拾起