通痹合剂乙酸乙酯部位调节活化T细胞双信号分子的机制研究  

Pharmacological mechanisms of ethyl acetate extraction of Tongbiheji for active T cell signaling molecules

作　　者：郭林凯[1] 陈光星[1] 刘丽娟[1] 彭麟钧[1] 谭选江[1] 廖黔华[1] 蒋川[1] 彭明欢[1] 

机构地区：[1]广州中医药大学金匮教研室,广东广州510405

出　　处：《细胞与分子免疫学杂志》2012年第3期265-267,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30873243);教育部新世纪优秀人才项目资助(2009年)

摘　　要：目的:明确通痹合剂抑制T淋巴细胞活化的活性部位,研究其对T细胞双信号分子的影响,阐明通痹合剂免疫调控作用的机制。方法:分离大鼠淋巴细胞,加入通痹合剂不同提取部位,ConA刺激48 h,流式细胞术(FCM)检测CD3阳性细胞表达CD71;加入不同浓度通痹合剂乙酸乙酯部位和甲氨蝶呤(MTX),ConA刺激48 h,FCM检测T细胞表达TCR、CD28和ICOS。结果:ConA刺激后CD3阳性细胞表达CD71显著上调至69.7%,通痹合剂乙酸乙酯部位(TBHJ)可抑制CD3+CD71+表达(32.5%);ConA刺激T细胞表达TCR、CD28和ICOS明显升高,与阴性对照组(Control)有非常显著性差异(P<0.001),TBHJ可明显抑制T细胞表达TCR、CD28和ICOS,尤其抑制ICOS作用较强,呈浓度依赖效应;MTX可明显抑制T细胞表达TCR、CD71。结论:TBHJ可明显抑制T淋巴细胞活化,其对T细胞活化的双信号分子途径均有下调作用,尤其以抑制ICOS明显,提示TBHJ可通过阻断CD28-ICOS第二信号途径,诱导免疫耐受。本研究为通痹合剂治疗类风湿性关节炎等自身免疫性疾病提供了实验依据。AIM： To confirm extracts of activity from Traditional Chinese Medicine TongBiHeJi,study effect on two signaling pathways of T cells and clarify the pharmacological mechanisms of TongBiHeJi.METHODS： Concanavalin（ConA） were added successively into rats lymphocytic culture with different extracts of activity from Traditional Chinese Medicine.After 24 hours,CD71 expression rate on rat T lymphocytes activated with ConA was analyzed by flow cytometry.TCR,CD28 and ICOS on T cells were detected after T lymphocytes of rat activated by ConA were cultivated with various EthylAcetate extraction of TongBiHeJi（TBHJ） and Methotrexate（MTX） for 48 hours.RESULTS： CD71 expression rate on rat T lymphocytes induced by ConA was increased to 69.7%.TBHJ inhibited the rate of CD3＋CD71＋ expression（32.5%）;ConA up-regulated TCR,CD28 and ICOS expression on T cells obviously.There was different between ConA and positive control significantly（P〈0.001）.TBHJ could down-regulate obviously TCR,CD28 and ICOS expression on ConA-activated T lymphocytes with Concentration-dependent,especially ICOS.MTX inhibited CD3＋CD71＋ and CD3＋TCR＋ expression also.CONCLUSION： TBHJ inhibited T cells activation by adjusting two signaling pathways.That implied TBHJ could block CD28-ICOS signaling molecules to induce immunological tolerance.This study provided an experimental basis for application of TongBiHeJi to treatment of rheumatoid arthritis.

关 键 词：通痹合剂乙酸乙酯部位 T淋巴细胞 TCR 

分 类 号：R392.12[医药卫生—免疫学]