奇壬醇对牛Ⅱ型胶原特异性淋巴细胞的体内和体外作用(英文)  

Effects of kirenol on bovine type Ⅱ collagen-induced rat lymphocytes in vivo and in vitro

作　　者：卢月[1] 肖娟[2] 吴再旺[1] 王喆明[1] 付宏征[3] 陈英玉[2] 钱瑞琴[1] 

机构地区：[1]北京大学医学部基础医学院中西医结合教研室,100191 [2]北京大学医学部人类疾病基因中心,100191 [3]北京大学医学部然与仿生药物国家重点实验室,100191

出　　处：《南方医科大学学报》2012年第1期1-6,共6页Journal of Southern Medical University

基　　金：Supported by National Key New Drug Creation Program of China(2012ZX09101);Natural Science Foundation of Beijing(7072043)~~

摘　　要：目的观察奇壬醇对牛II型胶原特异性淋巴细胞体内和体外的免疫调节作用,探讨奇壬醇对抗原特异性淋巴细胞的免疫抑制作用机制。方法将Wistar大鼠随机分为对照组、模型组、奇壬醇组和泼尼松龙组。除对照组外,均复制胶原诱导关节炎大鼠模型,造模当天开始给药。造模30 d后取脾脏和引流淋巴结,制备单细胞悬液,经体外培养48 h后,ELISA法检测培养上清中干扰素-γ(IFN-γ)、白介素-10(IL-10)、IL-4、肿瘤坏死因子-α(TNF-α)含量;在II型胶原诱导下,取正常大鼠淋巴结细胞、脾细胞与不同浓度的奇壬醇共同培养,检测胶原诱导的淋巴细胞增殖及凋亡。结果与模型组相比,奇壬醇组脾细胞培养上清中IFN-γ和TNF-α含量降低(P<0.05,P<0.01),IL-10和IL-4水平升高(P<0.05,P<0.01);淋巴结细胞培养上清中IFN-γ和TNF-α含量降低(P<0.05,P<0.001),IL-10和IL-4水平升高(P<0.05,P<0.001);体外实验中,奇壬醇抑制II型胶原特异性淋巴结细胞增殖,促进II型胶原特异性淋巴结细胞和脾细胞凋亡,均呈剂量依赖关系。结论奇壬醇可通过抑制II型胶原特异性淋巴细胞分泌促炎因子,促进胶原特异性淋巴细胞分泌抗炎因子,抑制胶原特异性淋巴细胞增殖和促进其凋亡,以多条途径发挥免疫抑制作用。Objective To investigate the effect of kirenol on bovine type II collagen（CII）-specific lymphocytes in vivo and in vitro,and explore the mechanism of kirenol-induced immunosuppression in antigen-specific lymphocytes.Methods Twenty-four Wistar rats were randomized into control group,collagen-induced arthritis（CIA） model group,kirenol group（2 mg/kg）,and prednisolone group（2 mg/kg）.After CII injection,the rats in the latter two groups received intragastric administration of kirenol and prednisolone for 30 days,and the spleens and draining lymph nodes of the rats were harvested to prepare single cell suspensions for measurement of the cytokine levels using ELISA.In the in vitro experiment,the lymphocytes from the control rats,with or without 20 μg/ml CII treatment in the presence of 0-80 μg/ml kirenol,were evaluated for cell proliferation and apoptosis using -thymidine incorporation and flow cytometry,respectively.Results Compared with those in CIA group,IFN-γ and TNF-α production was significantly reduced in splenocyte culture supernatant of kirenol group（P0.05 and P0.01,respectively）,and the level of IL-10 and IL-4 was up-regulated（P0.05 and P0.01,respectively）;IFN-γ and TNF-α secretion by the cultured lymph node cells（LNCs） significantly decreased（P0.05 and P0.001,respectively） and IL-10 and IL-4 production increased（P0.05,P0.001） in kirenol group.In the in vitro experiment,kirenol treatment caused obvious suppression of CII-induced LNC proliferation and dose-dependently induced antigen-specific apoptosis of the splenocytes and LNCs.Conclusion Kirenol treatment reduces pro-inflammatory cytokine secretion,increases anti-inflammatory cytokine production,inhibits cell proliferation and induces apoptosis of CII-specific lymphocytes in vitro,suggesting the potential of kirenol as an immunosuppressant.

关 键 词：奇壬醇 牛II型胶原 淋巴细胞 类风湿性关节炎 

分 类 号：R96[医药卫生—药理学]