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补肾中药防治去睾大鼠骨丢失的组织学机理研究成羿;顾庆玲;朱杭;黄海;陈红梅;章煜铭【期刊名称】《中国中西医结合外科杂志》【年(卷),期】2001(007)002【摘要】目的:探讨补肾中药防治去睾大鼠骨丢失的组织学机理.方法:选用10月龄大鼠行去睾术作为病理模型.分组为:正常组、病理组和补肾中药组.用药12周后处死,取腰1～2脊椎,作不脱钙骨切片,测定骨形态计量学各骨量参数和表面参数.各组参数作方差分析显著性检验.结果:①病理组骨量参数小于正常组(P＜0.05);表面参数显示:成骨参数减少、破骨参数增加、骨转换加快(P＜0.05).②补肾中药组骨量参数高于病理组(P＜0.05);表面参数得到逆转.同时可见部分成骨细胞特别活跃,其下类骨质高出周围静止表面.结论:补肾中药防治去睾大鼠骨丢失有三方面的作用机理:①促进骨形成和抑制骨吸收的双重作用;②降低骨转换,提高骨质量;③骨重建过程的短路效应.为临床防治老年性骨质疏松症提供了理论依据.【总页数】4页(P98-101)【作者】成羿;顾庆玲;朱杭;黄海;陈红梅;章煜铭【作者单位】杭州市中医院,;杭州市中医院,;杭州市中医院,;杭州市中医院,;杭州市中医院,;杭州市中医院,【正文语种】中文【中图分类】R28【相关文献】1.去睾大鼠骨丢失的组织学机理研究 [J], 成羿;朱杭;黄海2.雄性大鼠去睾后骨丢失及健脾中药和钙剂对其治疗作用的实验研究 [J], 成羿;朱杭;黄海3.补肾活血方药对去睾大鼠血清性腺激素及IL-6水平的影响 [J], 陈芝喜;刘庆思;邵敏;庄洪;陈津岩;李志强4.补肾中药防治去睾大鼠骨丢失的组织学机理研究 [J], 黄海;成羿;朱杭;陈红梅;章煜铭5.益肾健脾胶囊配合钙剂防治雄性大鼠去睾后骨丢失的组织形态计量学研究 [J], 成羿;朱杭;黄海;陈红梅;章煜铭(指导)因版权原因，仅展示原文概要，查看原文内容请购买。
NEURAL REGENERATION RESEARCH Volume 7, Issue 21, July 2012Cite this article as: Neural Regen Res. 2012;7(21):1611-1617.1611Siyun Niu ☆, M.D., Associate professor, Master’ssupervisor, School of Basic Medical Science, Hebei University, Baoding 071000, Hebei Province, ChinaSiyun Niu and Suru Kou contributed equally to this study.Corresponding author: Siyun Niu, School of Basic Medical Science, Hebei University, Baoding 071000, Hebei Province, China nsy1688@Received: 2012-02-15 Accepted: 2012-05-13 (N20111107001/WJ)Niu SY , Kou SR, Zhou XC, Ding L. Heshouwu decoction, a Chinese herb for tonifying kidney, ameliorates hypothalamic-pituitary- testicular axis secretion in aging rats. Neural Regen Res. 2012;7(21):1611-1617.doi:10.3969/j.issn.1673-5374.2012.21.002Heshouwu decoction, a Chinese herb for tonifying kidney, ameliorates hypothalamic-pituitary- testicular axis secretion in aging rats****☆Siyun Niu 1, Suru Kou 1, Xiaochun Zhou 2, Liang Ding 11School of Basic Medical Science, Hebei University, Baoding 071000, Hebei Province, China2Institute of Basic Medical Sciences, Chengde Medical College, Chengde 067000, Hebei Province, ChinaAbstractAn increasing amount of evidence demonstrates the anti-aging effect of Heshouwu in pill form. In this study, a subacute aging rat model was established by continuous intraperitoneal injection of D-galactose and treated with Heshouwu decoction (a Chinese herb for tonifying the kidney,comprising Heshouwu pill, Herba Epimedii, Radix Salviae Miltiorrhiae, and Poria). Heshouwu pill treated rats were the positive control group. Radioimmunoassay, immunohistochemical staining, and western blot assay showed hypothalamic gonadotropin-releasing hormone, hypothalamic substance P , and serum gonadotropin levels to be significantly increased in the model rats; the concentrations of hypothalamic β-endorphin, and serum levels of insulin-like growth factor 1 and testosterone were significantly decreased. 17β- and 3β-hydroxysteroid dehydrogenase expression in testicular tissue was also decreased. Intragastric administration of Heshouwu decoction at high (9.6 g/mL/100 g), medium (4.8 g/mL/100 g), and low (2.4 g/mL/100 g) doses, Heshouwu decoction pretreatment at a medium dose (4.8 g/mL/100 g), and Heshouwu pill (2.06 g/mL/100 g) significantly reversed these changes. Heshouwu decoction pretreatment and high-dose Heshouwu decoction had the greatest anti-aging effects. These experimental findings indicate that Heshouwu decoction can improve hypothalamic-pituitary-testicular axis secretion in a subacute aging rat model, and prevent and delay gonadal axis aging, with an effect superior to that of Heshouwu pill.Key WordsHeshouwu decoction; pituitary gonadal axis; aging; gonadotropin-releasing hormone; gonadotropin; hydroxysteroid dehydrogenase; hypothalamic-pituitary-testicular axis; neural regenerationResearch Highlights(1) Gonadotropin-releasing hormone, substance P , and gonadotropic hormone are significantly increased in the hypothalamic-pituitary-testicular axis of aging rats, whereas β-endorphin,insulin-like growth factor-1, testosterone concentration, 17β-hydroxysteroid dehydrogenase and 3β-hydroxysteroid dehydrogenase are significantly decreased. (2) Heshouwu decoction can reverse these phenomena and its effect is superior to that of Heshouwu pill.AbbreviationsGnRH, gonadotropin-releasing hormone; LH, luteinizing hormone; FSH, follicle-stimulating hormone; IGF-1, insulin-like growth factor 1; HSD, hydroxysteroid dehydrogenase1612INTRODUCTIONAging is an irreversible, natural phenomenon, andgonadal hormones may directly or indirectly regulate the aging process [1]. This study aimed to improve gonadal hormone levels, which are closely related to human aging. An increasing amount of evidence has demonstrated the anti-aging effect of Heshouwudecoction, and the underlying mechanism may depend on the following factors [2-3]: (1) improved antioxidantcapacity and regulation of blood lipids; and (2) influences on p53/pRb related protein expression in the aging pathway in testicular cells. However, the influence of Heshouwu decoction on hypothalamic-pituitary-testicular axis secretion and on the activity of the key enzymes in testosterone biosynthesis remains unclear.In the present study, Heshouwu decoction pretreatment and treatment was given to a D-galactose-induced rat aging model and hypothalamic-pituitary-testicular axis secretion was investigated in an attempt to understand the regulatory effect on gonadal axis secretion.RESULTSQuantitative analysis of experimental animalsEighty Sprague-Dawley rats, after 1 week of adaptive feeding, were divided randomly into seven groups. Normal group (n = 10) rats were injectedintraperitoneally with normal saline. For the model group (n = 10), subacute aging was induced by intraperitoneal injection of D-galactose. In themedium-dose Heshouwu decoction pretreatment group (n = 12), rats were pretreated with 4.8 g/mL/100 gHeshouwu decoction to observe the protective effect of this drug against D-galactose-induced subacute aging. In the Heshouwu decoction treatment groups (n = 12), following intraperitoneal injection of D-galactose, rats were injected with Heshouwu decoction at a high(9.6 g/mL/100 g), medium (4.8 g/mL/100 g), or low (2.4 g/mL/100 g) dose, or given Heshouwu in pill form (2.06 g/mL/100 g) suspension, to observe thetherapeutic effects on D-galactose-induced subacute aging. In the natural recovery group, model rats were allowed to recover spontaneously, to determine whether recovery from D-galactose-induced subacute aging can occur naturally over time. All 80 rats were involved in the final analysis.17β-hydroxysteroid dehydrogenase (HSD) and 3β-HSD expression in rat testicular tissueImmunohistochemical staining showed that 17β-HSD and 3β-HSD were expressed in a dispersed pattern inthe cytoplasm of rat Leydig cells. Semi-quantitative analysis by western blot assay demonstrated that17β-HSD and 3β-HSD protein expression in the model rats was significantly lower than that in the normal group (P < 0.01). Expression was increased to varying degrees in the Heshouwu decoction pretreatment group, theHeshouwu decoction treatment groups, and the Heshouwu pill group (P < 0.05). Levels in the Heshouwu decoction pretreatment group and the high-dose Heshouwudecoction treatment group were higher than those in the other groups (P < 0.05; Table 1, Figures 1 and 2). These symptoms were apparently reversed after intragastric administration of Heshouwu decoction at high (9.6 g/mL/100 g), medium (4.8 g/mL/100 g), and low (2.4 g/mL/100 g) doses, by Heshouwu decoction pretreatment (4.8 g/mL/100 g), and by Heshouwu pill (2.06 g/mL/100 g).Gonadotropin-releasing hormone (GnRH), substance P and β-endorphin changes in rat hypothalamus Radioimmunoassay showed that hypothalamic GnRH and substance P levels were significantly increased in the model rats compared with the normal group (P < 0.01), whereas β-endorphin was significantly decreased (P < 0.01). High-dose Heshouwu decoction andHeshouwu decoction pretreatment were superior to the other treatments (P < 0.05; Table 2).Serum levels of luteinizing hormone (LH),follicle-stimulating hormone (FSH), testosterone, and insulin-like growth factor 1 (IGF-1)Radioimmunoassay showed that serum LH and FSH levels in the model rats were significantly higher than those in the normal group (P < 0.01), whereastestosterone levels were significantly decreased (P < 0.01).Enzyme-linked immunosorbent assay demonstrated thatthe IGF-1 level in aging rats was significantly lower thanthat in the normal rats (P < 0.01). Serum LH and FSHlevels were decreased to varying degrees in theHeshouwu decoction pretreatment group, the Heshouwudecoction treated groups, and the Heshouwu pill treatedgroup, but IGF-1 and testosterone levels were increased(P < 0.01). Heshouwu decoction pretreatment and high-and medium-dose Heshouwu decoction were superior tolow-dose Heshouwu decoction and Heshouwu pill (P <0.05; Table 3).Figure 1 17β-HSD and 3β-HSD expression intesticular tissue (immunohistochemical staining,substance P method, optical microscope, × 400).17β-HSD (A–D) and 3β-HSD (E-H) were expressedpredominantly in the cytoplasm of Leydig cells.In the model group (A, E), 17β-HSD and 3β-HSDpositive products were stained light yellow.In the normal group (B, F), the Heshouwu decoctionpretreatment group (C, G), and the high-doseHeshouwu decoction group (D, H), the positiveproducts were brown, and their levels were greaterthan in the model group. (I) Negative control group.HSD: Hydroxysteroid dehydrogenase.A B CD E FG H IFigure 2 17β-HSD and 3β-HSD protein expression intesticular tissue.1: Model group; 2: normal group; 3: Heshouwu decoctionpretreatment group; 4: high-dose Heshouwu decoctiongroup; 5: medium-dose Heshouwu decoction group; 6:low-dose Heshouwu decoction group; 7: Heshouwu pillgroup; 8: spontaneous recovery group.17β-HSD and 3β-HSD protein expression was greatest inthe Heshouwu decoction pretreatment and high-doseHeshouwu decoction groups.HSD: Hydroxysteroid dehydrogenase.17β-HSD3β-HSDβ-actin1234567834 kDa43 kDa43 kDa1613DISCUSSIONThe D-galactose-induced subacute aging model is based on Senescence Metabolism Theory. The aging phenomena induced by D-galactose reflect the natural state of senescence within a short period of time, so this model is commonly used[4-5]. The drug is administered via intraperitoneal injection at a dose of 40-500 mg/kg per day for 20-60 days[6]. This study used intraperitoneal injection of D-galactose to produce a subacute aging model in the rat.Increasing evidence indicates the anti-aging effects of various single herbs and herbal compounds. Wanget al [7] demonstrated that saponins in Radix Ginseng, American Ginseng, and Radix Notoginseng can scavenge free radicals. Barbarum polysaccharide also exhibits an antioxidative capacity, with effects such as improving superoxide dismutase activity in the serum, heart, liver, and brain tissue in a D-galactose-induced aging mouse model, reducing malondialdehyde, and increasing serum and cardiac telomerase activity[8]. Weng et al [9] studied the influence of Ganodermasides A and B (two monomer compounds) in Ganoderma on the replication and survival of Saccharomyces cerevisiae, and found an anti-aging effect. Many Chinese herbal compounds, such as Apozem of Tremella and Wolfberry Fruit, Yangzhen Oral Liquid, and Yishuotiaozi Tablet, increase the activity of antioxidant enzymes, reduce malondialdehyde accumulation, and improve immunity. Vitality Reinforcing Prescription downregulates expression of the tumor suppressor gene p53 in spleen and liver cells of aging male mice at the transcription and translation levels. Jinguishenqi Pills are well known as antagonists of the DNA damage caused by cyclophosphamide. Liuwei Dihuang decoction can prolong the survival of Drosophila and increase telomerase activity in the brain and gonadal tissue of aging mice[10]. However, there is little evidence available on pituitary-testicular axis secretion, and the present study is the first demonstration of the action of Heshouwu decoction on pituitary-testicular axis secretion.Heshouwu decoction can regulate serum antioxidant capacity and lipid metabolic disorders[2]; through regulation of Rb/p16 and p53/p19/p21, this decoction also relieves cell conduction blockade, promotes the proliferation of ovarian and testicular cells, and inhibits apoptosis[3, 11-13]. Less attention had been paid to the regulation by Heshouwu decoction of testosterone. Our study showed that serum testosterone levels were significantly lower in aging rats than in the normal group, and Heshouwu decoction apparently improved serum testosterone levels in a dose-dependent manner. Pretreatment with Heshouwu decoction achieved optimal anti-aging effects.Testosterone synthesis and secretion are modulated by hypothalamic GnRH, peptide neurotransmitters, gonadotropins, and growth hormones secreted from the pituitary gland. Pituitary endocrine dysfunction is closely related to hypothalamic regulatory function[14], and the pulse and wave coordination of pituitary gonadotropic hormone and sex hormone secretion is decreased or absent in older males and females[15-16]. The hypothalamus-generated neuropeptides substance P and β-endorphin are also involved in GnRH synthesis and secretion[17]. By a feedback mechanism, testosterone can inhibit hypothalamic GnRH, pituitary FSH, LH, and IGF-1 levels. The role of Heshouwu decoction in this process has not yet been elucidated. In the present study, increased levels of hypothalamic substance P and GnRH, significantly decreasedβ-endorphin, and higher levels of serum FSH, LH, and IGF-1 were observed in aging rats and normal rats, and reversed by Heshouwu decoction. There is evidence that kidney-tonifying Chinese herbs regulate testosterone through hypothalamic GnRH secretion and release of the neurotransmitters β-endorphin and substance P, thereby affecting the secretion of FSH, LH, and IGF-1, or through the regulation of testosterone secretion by Leydig cells, providing negative feedback regulation of hypothalamic and pituitary function when gonadal axis secretion becomes abnormal as aging proceeds. Our findings support the conclusion that kidney-tonifying Chinese herbs regulate testosterone levels through the hypothalamic-pituitary axis.Leydig cell-secreted steroid dehydrogenases play an important regulatory role in testosterone synthesis and secretion. Various factors contribute to the influence on testosterone synthesis of 3β-HSD and 17β-HSD activity[18-20], but the role of Heshouwu decoction on3β-HSD and 17β-HSD synthesis and secretion in Leydig cells is unclear. This study observed 3β-HSD and 17β-HSD expression in testis tissue from aging rats and explored the regulatory effect of Heshouwu decoction.We found that 3β-HSD and 17β-HSD protein expression was significantly lower in aging rats than in normal rats, and Heshouwu decoction upregulated their expression in a dose-dependent manner. These experimental findings indicate that Heshouwu decoction can increase testosterone secretion in Leydig cells and regulate 3β-HSD and 17β-HSD levels. In summary, Heshouwu decoction can ameliorate hypothalamic-pituitary-testicular axis secretion in a subacute aging rat model, and prevent and delay gonadal axis aging, with an effect greater than that of Heshouwu pills.1614MATERIALS AND METHODSDesignA randomized, controlled animal experiment.Time and settingExperiments were performed from November 2009 to September 2010 at the Cell Biology Laboratory, School of Basic Medical Science, Hebei University, China.MaterialsAnimalsNinety clean, healthy male Sprague-Dawley rats, weighing 180-220 g, were provided by the Experimental Animal Center of Hebei Medical University, China (license No. 1005050).All experimental use of animals complied with the Guidance Suggestions for the Care and Use of Laboratory Animals, issued by the Ministry of Science and Technology of China[21].Chinese medicine compoundsHeshouwu decoction prescription: The decoction comprised Radix Polygoni Multiflori, Herba Cistanches, Achyranthes bidentata Blume, epimedium, Salvia miltiorrhiza, and tuckahoe (Traditional Chinese Medicine Hospital of Hebei Province, China) according to the ratio 3:2:3:2:5:3, immersed in eight times the volume of distilled water for 1 hour. The herbs were then decocted twice in simmer water, for 30 minutes each time. The decoction was condensed to high-, medium-, and low-dose liquids containing 2.4, 4.8, and 9.6 g/mL crude drug, respectively. All decoctions were stored at 4°C and rewarmed to 25-30°C before administration.Heshouwu pill prescription: The pill comprised Radix Polygoni Multiflori, Herba Cistanches, and Achyranthes bidentata Blume (Traditional Chinese Medicine Hospital of Hebei Province, China) according to the ratio 3:2:3, immersed in eight times the volume of distilled water for 1 hour. The herbs were then decocted twice in simmer water, for 30 minutes each time. The decoction was condensed to liquid containing 2.06 g/mL crude drug, which was stored at 4°C and re-warmed to 25-30°C before administration.Drug doses: All drug doses were calculated based on the adult dose[22]. Heshouwu decoction containing 2.4 g/mL crude drug was equivalent to a low adult dose, the medium dose was twice the low adult dose, and the high dose was four times the low adult dose. Heshouwu pill (2.06 g/mL) corresponded to the adult dose. Rats were given 1 mL/100 g at 15:00 each day for 60 consecutive days. MethodsEstablishment of subacute aging rat modelEighty 8-week-old, clean grade, healthy maleSprague-Dawley rats, weighing 180-220 g, were used in this study to produce the subacute aging model. Under aseptic conditions, D-galactose (Beijing Chemical Reagent Company, Beijing, China) was soaked in normal saline to prepare a 6% solution, which was given to the rats via intraperitoneal injection at a dose of 300 mg/kg per day for 60 consecutive days[3-6]. Other groups of rats were injected with normal saline solution for 60 days, once per day.Rat testis tissue and brain tissue samplesRats were killed under 10% chloral hydrate anesthesia, and blood samples of 2 mL were collected using a capillary pipette inserted into the medial orbital venous plexus and centrifuged at 800 ×g for 20 minutes. The supernatant was discarded and stored at -80°C. Following thoracotomy, cannulation was performed from the left ventricle to the ascending aorta for rapid infusion of cold saline (approximately 200 mL), then bilateral testicular tissue and brain tissue were rapidly removed. The left testis and the brain tissue were preserved in liquid nitrogen for 30 minutes and stored at -80°C. The right testis was fixed in paraformaldehyde, embedded in paraffin, and stained immunohistochemically.Radioimmunoassay for serum LH, FSH, and testosterone levelsA radioimmunoassay kit (Beijing Northern Biotechnology Research Institute, Beijing, China) was used to determine the serum levels of LH, FSH, and testosterone in strict accordance with the manufacturer’s instructions. Levels were calculated using a standard curve method.Enzyme-labeled immunosorbent assay for serum IGF-1 levelsAn enzyme-labeled immunosorbent assay kit (Wuhan Boster Biological Engineering Co., Ltd., Wuhan, Hubei Province, China) was employed to measure serum IGF-1 (rabbit anti-IGF-1 monoclonal antibody, goat anti-rabbit IgG; 78-5 000 pg/mL) in strict accordance with the manufacturer’s instructions. Levels were calculated using a standard curve method.Radioimmunoassay for hypothalamic GnRH, substance P, and β-endorphin levelsFive rats in each group were selected for harvesting of brain tissue. The hypothalamus was removed to the rear of the optic chiasma at the ventral side and weighed (approximately 30 mg). The specimens were then homogenized with 400 μL cell lysate, ground in an ice bath, and centrifuged at 4°C; the supernatant was1615discarded. GnRH, substance P, and β-endorphin were measured using a radioimmunoassay kit (PLA Navy Radioimmunoassay Center, China) according to the manufacturer’s instructions. Serum levels were calculated using a standard curve method.Immunohistochemical staining for 17β-HSD and3β-HSD expression in testicular tissueSlices were dewaxed to water and rinsed with PBS (0.01 M, pH 7.4) three times for 5 minutes each time, then incubated with 3% hydrogen peroxide-methanol solution at room temperature for 30 minutes to remove endogenous peroxidase activity. The specimens were incubated with normal goat serum at 37°C for30 minutes, after which the serum was absorbed with filter paper and the specimens were incubated with rabbit anti-17β-HSD (1:100) and anti-3β-HSD monoclonal antibody (1:50; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C overnight (20 hours), biotin-labeled secondary antibody (goal anti-rabbit IgG; Santa Cruz) at 37°C for 30 minutes and horseradish peroxidase-conjugated streptavidin (Beijing Zhongshan Jinqiao Biological Co., Ltd., Beijing, China) at 37°C for 30 minutes, and subjected to 3,3’-diaminobenzidine coloration (Beijing Zhongshan Jinqiao) under optical microscopy (Leica DM6000M, Wetzlar, Germany). Between each culture step, the specimens were rinsed with PBS three times for 5 minutes each time. After termination of the coloration, the specimens were counterstained with hematoxylin, dehydrated in an alcohol gradient, rendered transparent in xylene, and mounted with neutral gum. For negative controls, PBS was used instead of antibody, with the other steps being the same as those described above. The appearance of brownish yellow particles or fine particles in a diffuse distribution was considered a positive response.Western blot assay for 17β-HSD and 3β-HSD expression in testicular tissueTotal protein was extracted with a cell lysis kit and the total protein content of the specimens was determined with a bicinchoninic acid quantitative protein detection kit. Four per cent stacking gel and 10% separation gel were prepared, and 60 µg of testicular tissue protein was mixed with sampling buffer solution, boiled in a water bath for 5-10 minutes, and placed in the gel sample holes after natural cooling. Electrophoresis was performed under 80 V on the stacking gel and 100 V on the separation gel. The extracted proteins were transferred to polyvinylidene fluoride membrane using a water-bath electric transfer device at 4°C, at 30 V constant pressure overnight. The membrane was blocked with 5% skimmed milk powder and shaken gently for 1 hour at room temperature. The blocked polyvinylidene fluoride membrane was dried, placed into the hybridization bag, and incubated with rabbitanti-17β-HSD, 3β-HSD, and β-actin monoclonal antibodies (1:200; Santa Cruz) for 2 hours. The membrane was rinsed with TBST membrane three times for 10 minutes each time and oscillated gently, then dried and incubated with horseradish peroxidase-conjugated secondary antibody (goat anti-rabbit IgG antibody,1:2 000; Santa Cruz) at 37°C for 1 hour and rinsed in TBST three times for 10 minutes each time. Following polyvinylidene fluoride membrane coloration, Pierce chemiluminescent substrate solutions A and B were mixed for 15 minutes at room temperature and the scanned image was analyzed with a FUJI Mini-4000 (Fuji, Tokyo, Japan).Statistical analysisMeasurement data were expressed as mean ± SD using SPSS 16.0 software (SPSS, Chicago, IL, USA). Differences between groups were compared withone-way analysis of variance, and paired comparisons were made using the Student-Newman-Keuls test. P values less than 0.05 were considered significant.Funding:This study was supported by the Talent Introduction Fund of Hebei University, No. 2010-183; the Medical Science Special Fund of Hebei University, No. 2012A1005; the Key Project of Hebei Provincial Health Department, No. 20110151; and a grant from Hebei Provincial Administration of Traditional Chinese Medicine, No. 2011104.Author contributions: Siyun Niu had full access to the study concept and design, wrote the manuscript and managed the funds. Suru Kou was responsible for data collection and integration. Xiaochun Zhou performed statistical processing and data analysis. Liang Ding validated the study.Conflicts of interest:None declared.Ethical approval:This pilot was approved by the Experimental Animal Ethics Committee of Hebei University in China.REFERENCES[1] Bartke A. 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何首乌抗衰老作用研究近况何首乌抗衰老作用研究近况更新日期： 12-20 杨朝晔提要综述近十年来何首乌抗衰老作用方面的概况。
关键词何首乌；抗衰老；药理作用何首乌为蓼科植物何首乌Polygonum multilorum Thunb.的块根，系著名滋补中药，功能补肝肾，益精血，壮筋骨。
现代研究表明，何首乌具有增强免疫功能、降低血脂、抗动脉粥样硬化、延缓衰老、保肝、抗菌等作用［1］。
近十年来，关于何首乌抗衰老方面的研究十分活跃，现将其抗衰老作用近况概述如下。
1 增强免疫功能免疫机能下降学说是衰老发生的重要学说之一，免疫机能具有增龄性变化，即年龄增长则其免疫机能下降或紊乱，结果导致机体的衰老。
何首乌可以通过延缓免疫器官胸腺的衰老，提高机体非特异免疫和特异性免疫功能，从而延缓衰老。
1.1 提高免疫器官的重量：胸腺随增龄而逐渐萎缩，继而使机体胸腺依赖性免疫功能衰退。
何首乌可延缓胸腺退化与萎缩，使老年小鼠胸腺重量和体积明显增大，而对照组胸腺呈明显退化［2］，也可以提高老年大鼠“胸腺重/ 体重”比值［3］。
何首乌还能提高小鼠脾脏的重量［4］。
1.2 使胸腺组织学结构逆转：魏氏等通过何首乌对胸腺组织学结构研究发现［2，5］，对照组胸腺呈明显退化，何首乌组与对照组相比，皮质显著增厚，皮髓质分界清楚，细胞密度明显增大，其胸腺细胞一般呈圆形或卵圆形，并多见有丝分裂；而在明型上皮性网状细胞、巨噬细胞和交错突细胞内，线粒体嵴部分或全部或清晰可见，粗面内质网或轻度扩张、或呈扁平囊状；明型上皮性网状细胞、巨噬细胞和交错突细胞各自与胸腺细胞紧贴形成的花环也较常见。
表明何首乌有促进老龄小鼠胸腺超微结构明显逆转变化的作用。
1.3 增加胸腺核酸和蛋白质含量：何首乌能增加老年大鼠胸腺胞浆蛋白和核酸含量［3］，从而促进胸腺细胞增生，保护胸腺组织，延缓老年大鼠胸腺年龄性退化作用。
1.4 提高免疫细胞的功能：何首乌能使小鼠腹腔巨噬细胞吞噬指数明显上升［4］，提高机体的非特异性免疫功能。
2021年4月第23卷第2期April 2021 ,V〇1.23,No. 2湖北中医药大学学报Journal of Hubei University of Chinese Medicine【论著】首乌丸对衰老大鼠海马组织PPlY及病理形态的影响黄万凌u,徐荣鹏2,张超、汤琪、陈刚1(1.湖北中医药大学基础医学院，湖北武汉430065; 2.湖北中医药高等专科学校中医系，湖北荆州434020)摘要：目的通过观察衰老大鼠海马组织中蛋白磷酸酶ly(PPlY)的表达及海马组织病理形态的变化，探讨首乌丸对学习记忆能力的作用机制。
方法将SPF级SD雄性大鼠40只随机分为正常组、模型组、首乌丸组及维E组，除正常组外，其余各组均用D-半乳糖造成衰老模型，同时首乌丸组以首乌丸进行干预，维E组予维生素E。
6周后取海马组织，Real-timePCR检测海马组织中PPlymRNA的转录水平，HE染色观察海马组织病理形态学变化。
结果正常组PPlY的mRNA表达显著低于模型组(P<0.01),首乌丸组PPI y的mRNA表达低于模型组(P<0.05 ),首乌丸组海马组织中锥体细胞胞核固缩，形状不规则等情况较模型组明显减少，结论首乌丸可能通过降低PP1Y的mRN A表达,减轻海马组织的病理损伤来改善学习记忆功能。
关键词：衰老；首乌丸；海马；PPlY;病理形态；学习记忆功能中图分类号：R285.5 文献标识码：A doi：10.3969/j.issn. 1008-987x.2021.02. 01Effect of Shouwu Pills on PPly and pathomorphism in aging rats' hippocampusHUANG Wanling12,XU Rongpeng2,ZHANG Chao',TANG Qi',CHEN Gang1(1 .College of Basic Medicine,Hubei University of Chinese Medicine,Wuhan430065 ；2.Department of Chinese Medicine,Hubei College of Chinese Medicine,Jingzhou434020)Abstracts：Objective To investigate the mechanism of Shouwu Pills on learning and memory by observing the expression of protein serine /threonine phosphatase ly (P P ly) and pathologic changes in aging rats* hippocampus. Methods The rats were randomized divided into normal group,model group,Shouwu Pills group and vitamin E group,except normal group,the other groups were induced aging modelby D-galactose,at the same time,Shouwu Pills group was treated with Shouwu Pills, and vitamin E group was treated with vitamin E. After 6 weeks, the hippocampus was dissected, the expression of PPly mRN A was detected by real-time PCR,the pathologic changes in hippcampus were observed by HE stain method. Results The mRNA expression of PPly in normal group was significantly lower than that in model group (尸<0.01 )，and the mRN A expression of PPly in Shouwu Pills group was lower than that in model group (尸<0.05 )• Conclusion Shouwu Pills may improve learning and memory function by reducing the mRNA expression of PPly.Keywords：aging；Shouwu Pills；hippocampus；P P ly；pathomorphism；learning and memory function延衰增寿一直都是医学热点，随着老龄化社会的来 中不可忽视的普遍问题[n,如现今受到社会广泛关注的临，如何延缓衰老，减少各种老年性疾病的发生更是具 阿尔兹海默症，其主要的临床表现即进行性的记忆功有了重要的社会意义。
补肾化痰中药对烟熏大鼠慢支模型气管和肺组织AR表达的影响蒋士卿;王淑玲;闫爱华【摘要】目的:通过观察补肾化痰中药对烟熏大鼠慢支模型气管和肺组织中AR表达的影响,探讨补肾化痰中药防治慢支的作用机理.方法:将健康雄性Wistar大鼠72只随机均分为正常对照组、慢支模型组、气管炎咳嗽痰喘丸组、补肾化痰大剂量组、补肾化痰小剂量组和自然恢复组,每组12只,采用香烟烟雾吸入法建立雄性大鼠慢支模型,在模型复制成功后分别灌胃给药补肾化痰中药和气管炎咳嗽痰喘丸,治疗时间为30d,采用免疫组化SP法检测各组大鼠气管和肺组织中雄激素受体(AR)的表达,采用放免法测定各组大鼠血清睾酮(T)的含量.结果:与正常对照组比较,慢支模型组和自然恢复组大鼠气管和肺内支气管AR的表达水平均明显升高(P＜0.01,P＜0.05),血清T水平明显下降(P＜0.01);与慢支模型组比较,补肾化痰大、小剂量组和气管炎咳嗽痰喘丸组大鼠气管和肺内支气管中AR表达水平显著降低(P＜0.01,P＜0.05,P＜0.001),血清T水平明显升高(P＜0.05).结论:补肾化痰中药能使烟熏雄性大鼠慢支模型的气管和肺组织中AR的高水平表达降低,血清低水平T含量升高,提示补肾化痰中药调节雄激素及其受体水平可能是其治疗慢支的作用机理之一.【期刊名称】《中国中医基础医学杂志》【年(卷),期】2010(016)009【总页数】3页(P772-774)【关键词】雄性大鼠;慢性支气管炎;补肾化痰中药;气管炎咳嗽痰喘丸;雄激素受体(AR)、睾酮(T)、香烟烟雾吸入;免疫组化【作者】蒋士卿;王淑玲;闫爱华【作者单位】河南中医学院,河南,郑州,45000;郑州大学基础医学院,河南,郑州,450052;郑州大学基础医学院,河南,郑州,450052【正文语种】中文【中图分类】R285.5本研究采用香烟烟雾吸入法建立雄性大鼠慢支模型，采用免疫组织化学SP法检测大鼠气管和肺组织中雄激素受体(AR)的相对含量，探讨补肾化痰中药对雄性慢支大鼠气管和肺组织中AR表达的影响，为研究补肾化痰中药防治慢支的作用机制积累资料。
湖南农业大学课程论文学院：资源环境学院班级：农资一班姓名：金光日学号：200940409102 课程论文题目：何首乌的药用价值课程名称：中医药膳学评阅成绩：评阅意见：成绩评定教师签名：日期：年月日何首乌的药用价值学生：金光日1 药理作用1.1抗衰老作用许多学者认为，衰老动物体内积累大量脂质过氧化产物，并伴随超氧化物歧化酶活性的降低。
实验结果表明，何首乌可明显降低老年小鼠脑和肝组织丙二醛含量，增加脑内单胺类递质含量，增-4-强SOD活性，还能明显抑制老年小鼠脑和肝组织内单胺氧化酶-B 的活性，从而消除自由基对机体的损伤，延缓衰老和疾病的发生。
自由基学说认为，脂质过氧化物的生成和沉积可以引起一系列的衰老症状，因此脂质过氧化物的含量是评价衰老的主要指标之一。
何首乌提取物对小鼠皮肤脂质过氧化物的生成具有明显的抑制作用，说明何首乌具有延缓衰老的作用，可以作为良好的皮肤抗衰老化妆品添加剂，或是做成药膳食用。
此外，何首乌还能明显提高老年大鼠的外周淋巴细胞DNA损伤修复能力，通过抑制脑内单胺氧化酶-B 活性，影响生物体中枢神经递质的含量，从而调节中枢神经活动，延缓大脑的衰老。
1.2对免疫系统的影响制何首乌能拮抗免疫抑制剂氧化考的松或强的松龙引起的小鼠胸腺萎缩与退化作用，增加其胸腺、肾上腺、脾脏和胸腔淋巴结的重量，提高白细胞总数，促进胸腔巨噬细胞的吞噬功能，降低小鼠循环免疫复合物的含量。
免疫学认为，免疫功能的衰退与机体的老化密切相关，胸腺是免疫系统的中枢器官，能有效地维持机体的免疫功能。
何首乌能延迟随衰老出现的胸腺退化，可能是其延缓衰老、提高机体免疫力的重要机制。
此外，何首乌还能增加胸腺核酸和蛋白质含量，延缓老年大鼠胸腺年龄性退化作用，并能促进老龄小鼠胸腺超微结构明显逆转变化，使小鼠胸腔巨噬细胞吞噬指数明显上升，从而提高机体的非特异性免疫功能。
1.3降血脂及抗动脉粥样硬化作用实验研究表明，制何首乌的水提物可明显提高小鼠血清高密度脂蛋白胆固醇含量，降低TC水平。
一味何首乌，乌发、抗衰老，绝了！贪食伤肝现代研究表示，何首乌化学成分包含蒽醌类、二苯乙烯苷类、磷脂类、酚类和黄酮类等。
拥有抗衰老、提高免疫力、降血脂、抗动脉粥样硬化、抗炎、抗菌、抗癌、抗诱变等药理作用，毒理作用主要表现为肝脏毒性。
何首乌功能与作用何首乌为蓼科植物何首乌Polygonummultiflorumthunb.的干燥块根，主产于河南、湖北、广西、广东、贵州、四川、江苏等地，味苦、干、涩，性微温，归肝、心、肾经。
生首乌味甘、苦、性平，拥有解毒消痈、截疟、润肠通便的功能，用于疮痈、瘰疠、风疹瘙痒、久疟体虚、肠燥便秘等症[2]。
制首乌味甘、涩、微温，拥有补益精血、固肾乌须之功能。
现代研究表示，何首乌主要活性成分有蒽醌类、二苯乙烯类、磷脂类、黄酮类和酚类，可用于降血脂、抗衰老、提高免疫力、益智等1何首乌的化学成分[4]对采自泰山的何首乌化学成分进行了系统的研究，从中获取9个蒽醌类化合物，分别判定为大黄素（e modin）、大黄素甲醚（physcion）、拟石黄衣醇（迷人醇，fallacinol）、大黄素-8-甲醚（questin）、桔红青霉素（emodin-6,8-dimethylether）、ω-羟基大黄素（citreorosein）、大黄素-6,8-二甲醚、大黄素-8-O-β-D-吡喃葡萄糖苷（emodin-8-O-β-D-g lucopyranoside）、大黄素-8-O-(6-O-乙酰基)-β-D-吡喃葡萄糖苷[emodin-8-O-(6′-O-acetyl)-β-Dglucopyranoside]和大黄素甲醚-8-O-β-D-吡喃葡萄糖苷（physcion-8-O-β-Dglucopyranoside）。
不一样炮制工艺显示，总游离蒽醌含量：黑豆汁蒸片＞黑豆汁炖片＞清蒸片＞生片，总蒽醌含量：生片＞黑豆汁蒸片＞黑豆汁炖片＞清蒸片。
道的二苯乙烯苷单体成分有2,3,5,4-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（二苯乙烯苷），为什么首乌中的主要水溶性成分；2,3,5,4-四羟基二苯乙烯-2-O-(6′-O-α-D-吡喃葡萄糖)-β-D-吡喃葡萄糖苷；何首乌丙素（2,3,5,4-四羟基二苯乙烯-2,3-二-O-β-D-葡萄糖苷）；2,3,5,4-四羟基二苯乙烯-2-O-(6′-O-乙酰基)-β-D-葡萄糖苷等[3,6-8]。
