补脾益气中药对哮喘大鼠BALF中IL_5及GM_CSF含量的影响_杨玲
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黄芪对支气管哮喘大鼠支气管肺泡灌洗液及血清中NO、IL-5和炎症细胞的影响张雨;韦倩;伊鹏霏【摘要】目的:探讨黄芪对支气管哮喘模型大鼠血清及支气管肺泡灌洗液(BALF)中一氧化氮(NO)、白细胞介素5 (IL-5)和炎症细胞的影响,阐明黄芪对哮喘的治疗效果.方法:60只Wistar大鼠随机分为正常对照组、模型组、激素干预组及黄芪干预低、中、高剂量组,每组10只,除正常对照组外以卵白蛋白(OVA)致敏并吸入激发法制备大鼠哮喘模型,干预组分别于实验21 d给予黄芪2 mL(0.3g· 100g-1体质量)灌胃或普米克令舒2 mL雾化吸入,每天1次,连续14d,正常对照组与模型组大鼠给予等量生理盐水灌胃,每天1次,连续14 d.观察大鼠行为学变化,同时检测各组大鼠血清及BALF中NO、IL-5和炎症细胞的变化.结果:根据行为学观察大鼠哮喘造模成功.经治疗,大鼠行为状态转好,但未达到正常状态;黄芪干预组大鼠BALF和血清中NO和IL-5的含量较哮喘模型大鼠明显减少(P<0.01),BALF中炎症细胞总数、嗜酸性粒细胞百分比明显减少(P<0.01).结论:黄芪能够抑制哮喘模型大鼠BALF中IL-5和NO的增多,同时能够明显减少气道炎症细胞和嗜酸性细胞的活化和浸润,从而减轻呼吸道炎症,减轻哮喘症状或避免哮喘发作,达到防治哮喘的目的.【期刊名称】《吉林大学学报(医学版)》【年(卷),期】2011(037)006【总页数】4页(P1033-1036)【关键词】支气管哮喘;黄芪;一氧化氮;白细胞介素5;炎症细胞【作者】张雨;韦倩;伊鹏霏【作者单位】吉林大学和平校区门诊部,吉林长春130062;长春中医药大学附属医院心病科,吉林长春130021;吉林大学畜牧兽医学院兽医临床学教研室,吉林长春130062【正文语种】中文【中图分类】R285.5支气管哮喘是一种临床常见病,是由多种细胞(如嗜酸性粒细胞、肥大细胞、T淋巴细胞、中性粒细胞、呼吸道上皮细胞等)和细胞组分参与的呼吸道慢性炎症性疾病[1-3]。
补脾益气方药对哮喘大鼠嗜酸性粒细胞趋化因子表达的影响许珂玉;王艳杰;赵丹玉;柳春【期刊名称】《山东医药》【年(卷),期】2012(052)024【摘要】Objective To observe the intervention influence of traditional Chinese medicine (TCM) prescription for Reinfocing Pi Qi on the expression of eotaxin in asthmatic rat, and to explore its possible mechanism. Methods SPF male SD rats were randomly divided into normal control group (A group) . Asthmatic group (B group), normal saline group (C group) , therapeutic group of Dexamethasone (D group), therapaeutic group of TCM prescription for Reinforcing Pi Oi (E group). The asthmatic models were established by the classic way of OVA sensilization and excitation. Dexamethasone and normal saline were used as positive and negative control medicine. TCM prescription for Reinforcing Pi Qi were taken once a day in 1 mL/100 g for 14 days after seting up model in the rats of E group. The number of total cells, percentages of eo-sinophil (EOS), lymphocyte, neutrophil in bronchoaleolar lavage ( BALF) fluid were calculated. Pathological change of lung tissue were observed. The expression of eotaxin on level of mRNA and protein in lung tissue were detected by RT-PCR and Western blot assay. Results EOS number of BALF and expression of eotaxin in B group and C group were higher than those in A group (P < 0.01). EOS number of BALF and expression of eotaxin in Dgroup and E group were lower than those in B group (P < 0.01). However, there was no difference between D group and E group ( P > 0.05). Conclusion TCM prescription for Reinforcing Pi Qi can decrease the level of eotaxin in asthmatic rats, and alleviate inflammatory cells infiltration in lung, then inhibit airway inflamation.%目的观察补脾益气方药对哮喘大鼠肺组织嗜酸性粒细胞趋化因子(eotaxin)表达的干预作用.方法 SPF级雄性SD大鼠随机分为对照组(A组)、哮喘组(B组)、生理盐水治疗组(C组)、地塞米松组(D组)和补脾益气方药治疗组(E组).哮喘组通过卵蛋白(OVA)致敏、激发建立大鼠哮喘模型,以生理盐水和地塞米松分别做对照治疗.E组大鼠在复制哮喘模型后,按照1 mL/100 g 体质量给予补脾方药1次/d,共14 d.计数肺泡灌洗液细胞总数、嗜酸性粒细胞(EOS)、巨噬细胞、淋巴细胞和中性粒细胞数;光镜下观察肺组织病理学改变;采用RT-PCR和Western bloting检测肺组织eotaxin的表达.结果与A组比较,B、C组肺泡灌洗液EOS计数、eotaxin的mRNA和蛋白表达均增加(P<0.01),D、E 组较B组降低(P<0.01),D、E组间无差异(P>0.05).结论补脾益气方药可能通过降低哮喘大鼠体内eotaxin水平,减轻炎症细胞在肺组织局部浸润,抑制气道炎症.【总页数】4页(P4-6,后插1)【作者】许珂玉;王艳杰;赵丹玉;柳春【作者单位】辽宁中医药大学,沈阳110032;辽宁医学院;辽宁中医药大学,沈阳110032;辽宁中医药大学,沈阳110032;辽宁中医药大学,沈阳110032【正文语种】中文【中图分类】R289.5;R562.2【相关文献】1.补脾益气方药对哮喘大鼠支气管肺泡灌洗液中内皮素-1及肿瘤坏死因子的影响[J], 李刚;柳春;魏庆宇2.补脾益气方药对脾虚哮喘大鼠肺组织I к B/NF-к B信号途径的影响 [J], 柳春;王艳杰;赵丹玉3.补脾益气方对脾虚哮喘大鼠肺组织黏蛋白5AC表达的影响 [J], 王艳杰;柳春;赵丹玉;闵广艳;曹阳;郭隽馥4.补脾益气方药对哮喘大鼠Th1/Th2失衡的调节及机制 [J], 许珂玉;王艳杰;赵丹玉;杨如意;柳春5.丹参注射液对哮喘大鼠肺IL-13和嗜酸性粒细胞趋化因子表达的影响 [J], 王成国;薛克营;程立;李威;石明因版权原因,仅展示原文概要,查看原文内容请购买。
益气健脾中药对脾气虚大鼠神经肽Y、血管活性肠肽和丝裂原活化蛋白激酶14基因表达的影响目的观察脾气虚大鼠血清、小肠神经肽Y(NPY)和血管活性肠肽(VIP)含量,小肠组织丝裂原活化蛋白激酶14(Mapk14)mRNA表达的变化及益气健脾中药的干预效应,以揭示脾气虚证发生的内在机制。
方法受试动物随机分为空白组、模型组(7、14、21 d组)、益气健脾组,每组10只。
除空白组外,其余各组采用大黄法、力竭法及饥饿法复合法建立脾气虚证大鼠模型,造模同时益气健脾组每日给予四君子汤20 g/kg干预21 d,各组大鼠分时段采用放免法测定小肠、血清NPY和VIP含量,采用实时荧光定量PCR检测小肠组织Mapk14 mRNA表达水平。
结果与空白组比较,模型组大鼠血清及小肠组织NPY含量降低(P<0.05),VIP含量升高(P<0.05),小肠组织Mapk14 mRNA相对表达量升高(P<0.05),且以模型21 d组显著;与模型21 d组比较,益气健脾组大鼠血清及小肠组织NPY含量升高(P<0.05),VIP含量降低(P<0.05),小肠组织Mapk14 mRNA相对表达量降低(P<0.05)。
结论益气健脾中药具有调节脑肠肽NPY、VIP分泌及抑制小肠组织Mapk14 mRNA异常表达的作用。
标签:脾气虚证;神经肽Y;血管活性肠肽;丝裂原活化蛋白激酶;四君子汤;大鼠中医临床“脾虚证”病程中出现纳呆、腹胀、大便溏结不调等症状均为胃肠道生理功能紊乱所致。
研究表明,胃肠激素分泌紊乱是导致胃肠功能障碍和脾虚发生的原因之一[1]。
而丝裂原活化蛋白激酶(mitogen-activated protein kinase,Mapk)级联是细胞内广泛存在的丝/苏氨酸蛋白激酶超家族,是将细胞质的信号传递至细胞核并引起细胞核发生变化的重要物质,它与细胞凋亡、增殖、肿瘤发生以及氧化应激性肠上皮细胞损伤等有密切关系,是细胞内信号传递的重要途径[2-4]。
健脾益气方对肝癌模型大鼠的治疗作用及机制研究作者:王超岳紫晨音金萍蒋筱卓少元来源:《中国药房》2021年第19期中圖分类号 R285.5 文献标志码 A 文章编号 1001-0408(2021)19-2342-05DOI 10.6039/j.issn.1001-0408.2021.19.06摘要目的:探讨健脾益气方对二乙基亚硝胺(DEN)诱导的肝癌模型大鼠的治疗作用及机制。
方法:将80只雄性SD大鼠分为正常组、模型组、NOD样受体家族3(NLRP3)抑制剂组(MCC950,4.5 mg/kg)、胱天蛋白酶1(caspase-1)抑制剂组(VX-765,4.5 mg/kg)和健脾益气方低、中、高剂量组(5.25、10.5、21 g/kg),除模型组20只大鼠外(其中10只用于判断是否造模成功),其余每组10只。
除正常组大鼠腹腔注射生理盐水外,其余各组大鼠腹腔注射DEN(70 mg/kg)以复制肝癌模型。
造模成功后,正常组和模型组大鼠灌胃生理盐水,各抑制剂组大鼠腹腔注射相应药物,健脾益气方各剂量组大鼠灌胃相应药物,每日1次,连续4周。
末次处置后,观察大鼠肝组织病理学形态变化,检测大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的含量,检测大鼠肝组织中NLRP3以及细胞程序性坏死相关蛋白[衔接子凋亡相关斑点样蛋白(ASC)、caspase-1前体(pro-caspase-1)、受体相互作用蛋白激酶1(RIP1)、RIP3、混合系激酶区域样蛋白(MLKL)]的表达水平。
结果:与正常组比较,模型组大鼠的肝细胞可见不同程度的脂肪变性、细胞核增大和呈团块状,部分可见出血和坏死,并伴有增生灶和结节;其肝组织损伤指数和血清中TNF-α、IL-1β含量以及肝组织中NLRP3、ASC、pro-caspase-1、RIP1、RIP3、MLKL蛋白的表达水平均显著升高(P关键词健脾益气方;肝癌;NOD样受体家族3炎症小体;程序性坏死Study on Therapeutic Effects and Its Mechanism of Jianpi Yiqi Decoction on Liver Cancer Model RatsWANG Chao1,2,YUE Zichen1,YIN Jinping1,JIANG Xiao1,ZHUO Shaoyuan1(1. School of Basic Medicine, Guangxi University of TCM, Nanning 530200, China; 2. Guangxi Key Laboratory of Integrated Traditional Chinese and Western Medicine Translational Medicine for High Incidence Infectious Diseases, Guangxi University of TCM, Nanning 530200, China)ABSTRACT OBJECTIVE: To explore the therapeutic effects and its mechanism of Jianpi yiqi decoction on diethylnitrosamine (DEN) induced liver cancer model rats. METHODS: Totally 80 male SD rats were divided into normal group, model group, Nod-like receptor family 3(NLRP3) inhibition group (MCC950,4.5 mg/kg), caspase-1 inhibitory group (VX-765,4.5 mg/kg), Jianpi yiqi decoction low-dose, medium-dose and high-dose groups (5.25, 10.5,21 g/kg), with 10 rats in each group except for 20 rats in model group (10 of them were only used to judge whether modeling was successful). Rats in each group were intraperitoneally injected with DEN (70 mg/kg) to induce liver cancer model, except for the rats in normal group which were replaced by normal saline. After modeling, normal group and model group were given normal saline intragastrically; inhibitor groups were given relevant medicine intraperitoneally; Jianpi yiqidecoction groups were given relevant medicine intragastrically, once a day, for consecutive 4 weeks. After last administration, histopathological morphology of liver tissue was observed. The contents of serum inflammatory factors TNF-α an d IL-1β were detected. The expression of NLRP3 and programmed cell necrosis associated protein (ASC, pro-caspase-1, RIP1, RIP3 and MLKL) in liver tissue were detected. RESULTS: Compared with the normal group, the hepatocytes of model group showed varying degrees of steatosis, enlarged nuclei, lumpy,bleeding and necrosis, accompanied by proliferative foci and nodules. Liver tissue injury index,serum content of TNF-α and IL-1β as well as the protein expression of NLRP3, ASC, pro-caspase-1, RIP1, RIP3 and MLKL in liver tissue were significantly increased (PKEYWORDS Jianpi yiqi decoction; Liver cancer; Nod-like receptor family 3 inflammasome; Programmed necrosis肝癌是最常见的恶性肿瘤之一。
补脾益气中药对脾虚哮喘大鼠环核苷酸水平的影响杨月妍;朱晓明;王巍;朴仁范;魏庆宇【期刊名称】《中华中医药学刊》【年(卷),期】2008(26)3【摘要】目的:探讨补脾益气中药对脾虚哮喘大鼠环核苷酸水平的影响。
方法:以中医理论为指导,建立了大鼠脾虚哮喘病证结合模型。
观察补脾益气法对模型动物支气管肺泡灌洗液(BALF)中细胞总数、嗜酸细胞数(Eos)及BALF上清液中环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)含量及二者的比值的影响,以及补脾益气中药和β2-受体激动剂(β2-AA)对BALF细胞孵育液中cAMP、cGMP含量及二者比值的影响。
结果:与正常对照组比较,脾虚哮喘动物和哮喘动物均表现为Tot、Eos升高,cAMP含量降低,cGMP含量升高,cAMP/cGMP比值降低。
脾虚哮喘动物的变化更为明显。
补脾益气中药对脾虚哮喘和哮喘动物均可降低其Tot、Eos降低cGMP含量,升高cAMP含量和cAMP/cGMP比值。
BALF细胞孵育液中,补脾益气中药与β2-AA联合应用能更为显著的降低cGMP含量,升高cAMP含量和cAMP/cGMP比值。
结论:补脾益气中药可升高cAMP含量,降低cGMP含量,纠正cAMP/cGMP比值。
补脾益气中药与β2-AA合用药可加强抗哮喘作用。
【总页数】3页(P528-530)【关键词】哮喘;补脾益气;环磷酸腺苷;环磷酸乌苷;动物模型【作者】杨月妍;朱晓明;王巍;朴仁范;魏庆宇【作者单位】辽宁省中医药研究院药剂科,辽宁沈阳110034;沈阳军区第202医院变态反应科,辽宁沈阳110003;延边大学中医学院,吉林延边133000【正文语种】中文【中图分类】R285.5【相关文献】1.补脾益气方药对脾虚哮喘大鼠肺组织I к B/NF-к B信号途径的影响 [J], 柳春;王艳杰;赵丹玉2.补脾益气方对脾虚哮喘大鼠肺组织黏蛋白5AC表达的影响 [J], 王艳杰;柳春;赵丹玉;闵广艳;曹阳;郭隽馥3.培土生金对脾虚哮喘大鼠气道环核苷酸水平的影响 [J], 孙晓玲;闫英海;张庆荣4.补脾益气中药对哮喘大鼠模型BALF中IL-5GM-CSF TNF-α和sIL-2R的影响[J], 杨月妍;魏庆宇;杨春山;朱晓明;谭雯;王巍因版权原因,仅展示原文概要,查看原文内容请购买。
益气合剂对免疫受抑大鼠免疫功能的影响马锦;杨如意【期刊名称】《青海医学院学报》【年(卷),期】2016(037)001【摘要】Objective To study the effect of YiQi Mixture on immune function of immunosuppressed rats. Methods The immunosuppressed model was established by intraperitoneal injecting of cyclophosphamide in rats.ser-um IgG,IgM and IgA,IL_2 production capacity,spleen index and thymus index were detected afteradministrating YiQi Mixture.Results SerumIgG,IgM,IgA and IL_2,the spleen index and thymus index in therapeutic group of YiQi Mixture were raised compared those with the model group.Condusion The YiQi Mixture can improve the im-mune function of immunosuppressed rats.%目的:研究益气合剂对免疫受抑大鼠的免疫调节作用。
方法腹腔注射环磷酰胺建立免疫受抑大鼠模型。
测定模型大鼠血清 IgG、IgA、IgM 含量,IL_2生成能力,胸腺指数与脾脏指数等免疫指标,评价益气合剂免疫调节作用。
结果益气合剂组大鼠血清 IgG、IgM、IgA 的含量, IL_2的生成能力及胸腺指数、脾脏指数与模型组比较明显升高(P﹤0.05)。
结论益气合剂能增强免疫受抑大鼠的免疫功能。
补脾益气中药对哮喘大鼠BALF 中IL -5及GM -CSF 含量的影响杨玲1,朱晓明2,王磊2,薛丹2,魏庆宇2(1.解放军309医院旃坛寺门诊部,北京100034;2.沈阳军区第202医院,辽宁沈阳110003)摘要:目的:探讨白细胞介素-5(IL -5)及粒细胞-巨噬细胞集落刺激因子(GM -CSF )在哮喘发病过程中的作用及补脾益气中药对其含量的影响。
方法:24只大鼠随机分成3组,每组8只,即:正常对照组、哮喘模型组和中药组。
采用卵蛋白、灭活百日咳杆菌疫苗和氢氧化铝干粉配制的溶液腹腔注射致敏以及卵蛋白雾化吸入激发制备大鼠哮喘模型;正常对照组用等量生理盐水替代抗原液;中药组则胃饲补脾益气中药(加味玉屏风散)水溶液。
观察支气管肺泡灌洗液(BALF )中IL -5及GM -CSF 含量的变化。
结果:与正常组比较,模型组BALF 中IL -5及GM -CSF 含量均显著增高(均P <0.05或P <0.01),尤其是IL -5增高更显著;相关分析表明,GM -CSF 与IL -5、呈显著正相关(r 1=0.519,r 2=0.521,均P <0.05)。
补脾益气中药能降低BAFL 中IL -5及GM -CSF 的含量(P <0.05或P <0.01)。
结论:IL -5、GM -CSF 在哮喘大鼠的气道炎症细胞浸润过程中起重要作用,可能加速了哮喘非特异性气道炎症的发生和发展;GM -CSF 与IL -5之间存在密切的联系,可能对嗜酸粒细胞的聚集、迁移、活化和黏附有协同作用。
补脾益气中药(加味玉屏风散)可以通过降低细胞因子水平来改善气道的炎症状况。
关键词:补脾益气中药;白细胞介素-5;粒细胞-巨噬细胞集落刺激因子(GM -CSF );哮喘动物模型中图分类号:R256.12文献标识码:B 文章编号:1000-1719(2011)11-2291-02Effect of Prescription and Medicine of Strengthening Spleen and BenefitingVital Qi on Content of IL -5and GM -CSF in BALF of Asthma Rat YANG Ling 1,ZHU Xiao-ming 2,WANG Lei 2,XUE Dan 2,WEI Qing-yu 2(1.Department of Zhantanshi Clinic ,No.309Hospital of PLA ,Beijing 100034,China ;2.Department of Allergy Diagnosis ,No.202Hospital of PLA ,Shenyang 110003,Liaoning ,China )Abstract :Objective :To explore the role of IL -5and GM -CSF in the course of asthmatic incidence ,and the effect of theprescription and medicine of strengthening spleen and benefiting Vital Qi on content of both of them.Methods :Twenty -four male rats were randomly divided into three groups :normal control group ;model group ;Chinese Herbal group.Every group has eight rats.A sensitized asthmatic rat model was obtained by intraperitoneal injection egg albumin 、inactivated Bordetella pertussis vac-cine and aluminum hydroxide dry flour of prepared solution ,and provocation by atomization breathe in egg albumin.Normal control group was supplied with partes aequales physiological saline to substitute antigen liquid ;Chinese Herbal group was given prescrip-tion and medicine of strengthening spleen and benefiting Vital Qi (Add Ingredients Jade Screen Powder.The change of content of IL -5and GM -CSF in broncho -alveolar lavage fluid (BALF )were observed.Results :Compared with normal control group ,Model group BALF has dramatic growth of the content of IL -5and GM -CSF (P <0.05in all or P <0.01),especially the IL -5.Correlation analysis showed that GM -CSF and IL -5were significantly direct correlated (r 1=0.519,r 2=0.521,P <0.05in all ).The prescription and medicine of strengthening spleen and benefiting Vital Qi can decrease the contents of IL -5and GM -CSF in BALF (P <0.05or P <0.01).Conclusion :IL -5and GM -CSF play an important role in the course of the asthmatic rat `s air way inflammatory cell infiltration.They may accelerate the emergence and development of the nonspecific airway inflamma-tion of asthma.There exists close link between IL -5and GM -CSF ,and it is expected that this has synergistic action on the ag-gregation ,migration ,activation and adherence of the eosinophil (EOS ).The prescription and medicine of strengthening spleen and benefiting Vital Qi (Add Ingredients Jade Screen Powder )can improve the airway ’s inflammation state through depressing the level of cell factor.Key words :Chinese herb for strengthering spleen and benefiting Qi ;IL -5;GM -CSF ;Asthma ;Anima model 收稿日期:2011-03-26基金项目:国家医药技术创新博士基金(96-901-06-15)作者简介:杨玲(1954-),女,主任医师,硕士,研究方向:耳鼻咽喉科临床。
通讯作者:魏庆宇(1964-),吉林长春人,主任医师,硕士研究生导师,博士,研究方向:过敏疾病的基础及临床。
细胞因子网络失调,可能是支气管哮喘(以下称哮喘)发病的分子生物学基础之一。
因此弄清各种细胞因子、炎性递质的功能及其相互关系,是研究哮喘气道炎症机理的重要课题。
本文观察了哮喘大鼠BALF 中IL -5及GM -CSFR 含量的变化,目的是探讨这两种细胞因子在哮喘发病中的协同作用,同时观察了补脾益气中药加味玉屏风散对它们的影响,探索中药治疗哮喘的理论基础,提供客观依据。
现报道如下。
1材料及方法1.1哮喘大鼠动物模型的复制[1-3]雄性Wistar 大鼠24只,体重160 250g ,随机分成生理盐水对照组(对照组)、哮喘发作组(哮喘组)和加味玉屏风散组(中药组)。
在实验的第1天、3天、5天大鼠腹腔注射抗原液1mL(含卵蛋白100mg,灭活百日咳杆菌疫苗5ˑ109个,氢氧化铝干粉100mg)致敏,对照组则用等量的生理盐水替代抗原液。
中药组给予加味玉屏风散(黄芪、白术、防风、冬虫夏草等)制成的100%浓度的药液,每日予2mL胃饲。
2周后用超声雾化器向自制体描箱内喷雾1%卵蛋白,吸入20min激发。
以后隔日定时激发1次,共3次,记录激发前后10min的呼吸曲线。
对照组则用生理盐水代替抗原液以同样方法激发和记录。
所有大鼠均经呼吸曲线描记及肺组织病理学检查,7天后按设计取材结果:连续抗原激发后哮喘组大鼠即时表现为呼吸急促,严重者呼吸减慢或节律不整,四肢瘫软,行动迟滞或俯伏不动,反应迟钝;连续激发后对照组大鼠RR和VT均无明显变化。
而哮喘组激发前VT即逐日下降,至第6天下降近50%(P<0.01),激发后下降更显著,病理组织学改变;对照组支气管和肺泡结构均正常。
哮喘组细、小支气管壁和伴行动脉周围有较多嗜酸细胞、浆细胞和淋巴细胞浸润。
平滑肌增厚,管腔缩窄,内有黏液栓。
部分肺泡壁变薄或断裂,融合成肺气肿。
上述结果符合哮喘的病理改变。
1.2实验方法1.2.1取材(1)腹腔注射1.5%戊巴比妥钠(0.3mL/100g)麻醉后,开胸气管插管,用生理盐水10mL注入气管肺组织内,反复抽吸3次,将抽取的肺泡灌洗液(BALF)放入离心管中离心10min(1000r/ min,离心半径10cm),取上清液用于检测IL-5及GM-CSFR的含量。
1.2.2方法及试剂(1)IL-5的测定:采用双抗体夹心免疫法(ELISA)试剂盒(美国Endoger公司产品),按说明书进行。
(2)GM-CSF的测定:采用双抗体夹心免疫法(ELISA)试剂盒(第四军医大学细胞因子试剂盒),按说明书进行。