系统性红斑狼疮患者外周血T细胞Rho激酶活性检测及其意义  

Abnormal activity of Rho kinase in peripheral blood T cells from patients with systemic lupus erythematosus

作　　者：黄红维[1] 许韩师[1] 詹钟平[1] 梁柳琴[1] 杨岫岩[1] 叶玉津[1] 邱茜[1] 

机构地区：[1]中山大学附属第一医院风湿免疫内科,广州510080

出　　处：《中华风湿病学杂志》2009年第12期833-836,共4页Chinese Journal of Rheumatology

基　　金：基金项目：国家自然科学基金（30671951）;教育部出国留学回国人员启动基金（2006-331）

摘　　要：目的研究系统性红斑狼疮（SLE）患者外周血T细胞中Rho激酶（ROK）活化情况，并进一步探讨其临床意义。方法收集SLE患者30例，健康人12名和类风湿关节炎（RA）患者8例作为对照组。外周血T细胞分离采用RosettSepT细胞提取试剂盒提取，ROK活性用磷酸化MYPT1蛋白表达来表示，磷酸化MYPT1蛋白检测采用免疫印迹法，T细胞增殖采用四甲基偶氮唑蓝比色（MTT）法，细胞因子水平采用酶联免疫吸附试验（ELISA）检测。结果新鲜分离的SLE患者外周血T细胞ROK活性均显著高于健康对照组和RA组；SLE患者T淋巴细胞在体外培养24h后ROK活性与健康对照组差异无统计学意义；ROK特异性抑制剂Y27632显著抑制SLE患者T淋巴细胞增殖以及分泌白细胞介素（IL）-6，但对IL-10分泌无显著影响。中、重度病情活动的SLE患者外周血T淋巴细胞ROK活性显著高于轻度活动者（P均〈0．05）。SLE患者关节炎、神经精神症状和蛋白尿表现阳性组T淋巴细胞ROK活性显著高于上述临床表现阴性组。结论SLE患者T淋巴细胞存在ROK信号活化异常，并且可能与T淋巴细胞活化、狼疮疾病活动性和某些脏器损害有关。Objective To investigate the activity of Rho kinase （ROK）, a cytoplasmic kinase, which is known to play an important role in immune regulation, in peripheral blood T ceils of SLE patients. Methods T cells were isolated by RosettSep T cell purification kit. ROK activity was assessed by Western blot. T cell proliferation and cytokine production was examined by MTT test and ELISA, respectively. Results Compared with healthy controls and rheumatoid arthritis patients, the activity of ROK in ex vivo T cells from SLE patients was significantly increased. Furthermore, T cells allowed to ＂rest＂ in vitro for 24 hours in culture medium showed a reversal of the changes in activity of ROK. Proliferation and secretion of IL-6 in cultured T cells from SLE patients were inhibited by Y27632, a specific inhibitor of ROK, however, IL-10 secretion was not affected. There is positive correlation between ROK activity of SLE T cell and SLE disease activity index （SLEDAI）. The ROK activities of T cells from SLE patients with arthritis, neuropsychiatrie symptoms and clinical renal injury were significantly higher than those patients who did not have above involvement, respectively. Conclusion The results indicate that ex vivo T cells from patients with SLE have increased activity of ROK, and suggest that alterations of ROK activity observed in SLE T cells may contribute, at least in part, to the pathogenesis of SLE in disease activity and tissue injury.

关 键 词：红斑狼疮 系统性 T淋巴细胞 信号转导 RHO激酶 

分 类 号：R686[医药卫生—骨科学]