青藤碱对T淋巴细胞活化增殖的影响  

Effect of Sinomenine on Activation and Proliferation of T Lymphocytes

作　　者：陈光星[1] 李晓娟[1] 刘清平[1] 刘晓玲[1] 黄可儿[1] 陈纪藩[1] 

机构地区：[1]广州中医药大学第一附属医院风湿专科,广东广州510405

出　　处：《广州中医药大学学报》2008年第5期425-428,共4页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金重点资助项目(编号:39930230);广东省中医药管理局课题资助(编号:102019)

摘　　要：【目的】观察青碱藤(SINO)对T淋巴细胞增殖和活化的影响及其细胞免疫调控机制。【方法】培养Jurkat细胞,分别加入不同浓度的SINO(终浓度分别为1、0.5、0.1 mmol/L),采用四甲基偶氮唑盐(MTT)法检测细胞增殖率;培养Jurkat细胞,分别设空白对照组,甲氨喋呤(MTX)阳性对照组(剂量为5 mg/L),不同浓度的SINO组(终浓度分别为1、0.5、0.1 mmol/L),采用流式细胞术检测各组细胞周期。无菌分离Wistar大鼠肠系膜淋巴结淋巴细胞,以1、0.5、0.1 mmol/L SINO预培养60 min后,分别加入刀豆蛋白(ConA,终浓度为10 mg/L)或佛波醇酯(PDB,终浓度为0.20μmol/L)+离子霉素(ionomycin,终浓度为1 mg/L)继续培养48 h,以抗大鼠CD3和CD71单抗标记后,采用流式细胞术检测各组CD3+CD71+表达情况。【结果】不同浓度SINO呈浓度依赖性抑制淋巴细胞增殖,且主要抑制Jurkats细胞G0/G1期,MTX作用于S期;ConA或PDB+ionomycin刺激可显著升高CD3+CD71+表达(均P<0.01),1、0.5 mmol/LSINO可显著抑制ConA刺激下的CD3+CD71+表达(P<0.01),1 mmol/LSINO可显著抑制PDB+ionomycin刺激下的CD3+CD71+表达(P<0.05)。【结论】SINO可抑制T淋巴细胞活化和增殖,将细胞阻断于G0/G1期,其作用可能与下调T细胞转铁蛋白受体的表达,减少细胞对铁离子摄入有关。Objective To observe the effect of sinomenine （SINO） on the proliferation and activation of T lymphocytes and to explore its cellular immune regulatory mechanism. Methods Jurkats cells were cultured with various concentrations of SINO （ at final concentrations of 1,0. 5 and 0. 1 mmol/L）, and the proliferative rate was detected by methyl thiazolyl tetrazolium （MTT） assay. The cultured Jurkats cells were allocated to blank control group, methotrexate （ MTX, 5 mg/L） group, and SINO groups （ at final concentrations of 1, 0. 5 and 0. 1 retool/ L）, and the cell cycle was examined in different groups after the culturing. Lymphocytes extracted from Wistar rats mesenteric lymph nodes under aseptic condition were pre-cuhured with SONO （ at final concentrations of 1,0.5 or 0. 1 mmoL/L） for 60 min, and then were stimulated by concanavalin （ ConA, at final concentration of 10 mg/L） or phorbol estersil （ at final concentration of 0. 20μmol/L） ＋ ionomycin （at final concentration of 1 mg/L） for another 48 hours. After marked by monoclonal antibody of CD3 and CD71, the expression of CD3＋ CD71＋ lymphocytes was observed by flow cytometry. Results SINO inhibited the lymphocytes proliferation in a concentration-dependant manner, and particularly had an inhibition on Jurkats cells at G0/G1 phase; MTX had an inhibition on S phase. The stimulation of ConA and phorbol estersil ＋ ionomycin obviously increased the expression of CD3＋ CD71＋ lymphocytes （P 〈 0. 01 ） ; SINO at the concentrations of 1 and 0. 5 mmol/L inhibited the expression of CD3＋CD71＋ lymphocytes after ConA stimulation, and SINO at the concentration of 1 mmol/L inhibited the expression of CD3＋ CD71＋ lymphocytes after stimulation of phorbol estersil ＋ ionomycin （P 〈0. 05）. Conclusion SINO can inhibit the activation and proliferation of T lymphocytes, and block the cells at G0/G1 phase. Its mechanism may be related with the down-regulation of transferring receptor of T lymphocytes and with the decrease of i

关 键 词：青藤碱/药理学 关节炎 类风湿/中药疗法 T淋巴细胞 细胞培养 

分 类 号：R285.5[医药卫生—中药学]