地塞米松体内及体外诱导系统性红斑狼疮患者CD4^+ CD25^- T淋巴细胞凋亡的作用  

The role of dexamethasone-induced systemic lupus erythematosus patients,CD4^+ CD25^-lymphocyte apoptosis in vivo and in vitro

作　　者：宋维亚[1] 李向培[1] 厉小梅[1] 马艳[1] 王俐[1] 俞宁[1] 曾小峰[2] 

机构地区：[1]安徽医科大学附属省立医院风湿免疫科,合肥230001 [2]中国协和医科大学北京协和医院风湿免疫科,北京100730

出　　处：《安徽医科大学学报》2010年第4期518-522,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家"十一五"科技支撑计划(编号:2008BAI59B02);国家自然科学基金(编号:C080701);安徽省自然科学基金(编号:090413129);安徽省临床医学重点学科应用技术(编号:05A008)

摘　　要：目的通过体内及体外途径研究系统性红斑狼疮(SLE)患者在应用地塞米松前后外周血CD4+ CD25- T淋巴细胞凋亡率的变化,探讨影响SLE患者经激素治疗后CD4+CD25+调节性T淋巴细胞(Treg)比率上调的机制。方法应用AnnexinV检测试剂盒及流式细胞仪检测未经激素治疗的10例SLE患者经地塞米松治疗0、1、3、7d后外周血单个核淋巴细胞(PBMC)中CD4+ CD25- T淋巴细胞的早期凋亡率;结合细胞培养技术检测未经激素治疗时SLE患者PBMC在体外经地塞米松刺激培养1、3、7d后CD4+ CD25- T淋巴细胞的早期凋亡率。结果地塞米松治疗后与未治疗之前相比,SLE患者PBMC中CD4+ CD25- T淋巴细胞早期凋亡率升高(P<0.05),治疗0、1、3、7d的凋亡率分别是0.21±0.03、1.39±0.22、1.57±0.11、1.83±0.31;而CD4+ CD25+ Treg细胞早期凋亡率无明显变化(P>0.05),治疗0、1、3、7d的凋亡率分别是0.44±0.06、0.47±0.04、0.43±0.07和0.35±0.07;未治疗的SLE患者PBMC经过体外地塞米松刺激培养后CD4+ CD25- T淋巴细胞早期凋亡率升高(P<0.05)。体外培养1、3、7d的凋亡率分别是2.02±0.29、2.50±0.59和3.72±0.36;未治疗的SLE患者PBMC经过体外地塞米松刺激培养后,CD4+ CD25- T淋巴细胞早期凋亡率随激素剂量的增加而上升(P<0.05)。体外培养7d后,DMX剂量为0、5、10、25μg时凋亡率分别为0.60±0.26、1.51±0.69、2.20±0.80和3.65±0.72;SLE患者CD4+ CD25- T淋巴细胞早期凋亡率与抗ds-DNA抗体、抗Sm抗体、IgG、IgA、IgM、抗SSA抗体、抗SSB抗体、C3、C4等均无相关性(P>0.05),与SLEDAI评分呈负相关(P<0.05)。结论 SLE患者经过地塞米松治疗后外周血CD4+ CD25- T淋巴细胞的凋亡增加并与剂量相关,引起外周血CD4+ CD25- T淋巴细胞数量的减少,可能是导致CD4+ CD25+ Treg细胞比例相对增加的原因。Objective To investigate the up-regulation mechanism of affecting Treg cells ratio in systemic lypus erythematosus（SLE） patients undergone glucocorticoid therapies,through the research of in vivo and in vitro observing changes of CD4^＋CD25^-T lymphocytes apoptosis in peripheral blood of SLE patients before and after glucocorticoid therapies.Methods Early stage CD4^＋CD25^-T lymphocytes apoptosis rate were tested with AnnexinV detection kit and flow cytometer in 10 cases of SLE patients,who had not been glucocorticoid therapies before and the detection were undertaked with dexamethasone therapies at 0,1,3 and 7 days.Cell cultivation technology was also adopted to test CD4^＋CD25^-T lymphocytes early apoptosis rate when dexamethasone stimulation were applied to peripheral blood mononuclear cells （PBMC）in vitro for 0,3 and 7 days.Results After dexamethasone therapies,SLE patients′ PBMC CD4^＋CD25^-T lymphocytes early apoposis rate rose obviously as compared with before the treatment（P〈.05）.The apoptosis rates were 0.21±0.03,1.39±0.22,1.57±0.11 and 1.83±0.31 at 0,1,3 and 7 days after treatment respectively.There was no significant change in early apoptosis rate for CD4^＋CD25^＋ Treg（P〈.05）.The apoptosis rates for 0,1 day,3 day and 7 day treatment were 2.02±0.29,2.50±0.59 and 3.72±0.36 repectively.After in vitro dexamethasone cultivation of untreated SLE patients′ PBMC,CD4^＋CD25^-T lymphocytes early apoptosis rate rose（P〈.05）.The apoptosis rates of vitro cultivation at 1,3 and 7 day were 2.02±0.29,2.50±0.59 and 3.72±0.36 respectively.After in vitro dexamethasone cultivation of untreated SLE patients′ PBMC,CD4^＋CD25^-T lymphocytes early apoptosis increased with the increase of Hormone dose（P〈.05）.Cultured in vitro at 7 days,apoptosis rates were 0.60±0.26,1.51±0.69,2.20±0.80 and 3.65±0.72 with the corresponding dose of DMX were 0,5,10 and 25 μg.Early apoptosis of SLE patients′ CD4^＋CD25^-effector T lymphocyte was not correlated to anti-ds-DNA antibodie

关 键 词：地塞米松 T淋巴细胞 细胞凋亡 红斑狼疮 系统性/化学诱导 

分 类 号：R593.241[医药卫生—内科学] R977.1[医药卫生—临床医学]