狼疮定对SLECD4^+T细胞凋亡及其GRα表达的作用  

Regulating Effects of Langchuangding on the Apoptosis and Expressions of GRα of CD4^+T Cells in SLE

作　　者：张莉[1] 孙静[2] 沈洁[3] 杨燕青[3] 何昱君[3] 范永升[3] 谢志军[3] 

机构地区：[1]杭州市中医院,杭州310007 [2]浙江中医药大学附属第二医院 [3]浙江中医药大学基础医学院

出　　处：《浙江中医药大学学报》2016年第6期423-430,共8页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金项目(81273680);高等学校博士学科点专项科研基金(20123322120001)~~

摘　　要：[目的]明确狼疮定(LCD)是否通过影响糖皮质激素受体α(glucocorticoid receptor alpha,GRα)的表达来增强糖皮质激素诱导激素不同敏感型系统性红斑狼疮(Systemic lupus erythematosus,SLE)CD4^+T细胞的细胞凋亡作用。[方法]筛选符合入选标准的激素抵抗型和敏感型的SLE患者,采集外周血并立即分选出CD4^+T细胞,分别随机分为3组,空白组加空白血清,地塞米松(Dexamethasone,DXM)组加空白血清和10μmol·m L-1 DXM,LCD+DXM组加中药含药血清2.5m L和10μmol·m L-1 DXM。分别药物处理24h后收集各组细胞,流式细胞术检测细胞凋亡,免疫荧光技术检测细胞内GRα的表达。[结果]共纳入58例符合标准的SLE患者,其中激素抵抗型25例,激素敏感型共33例。取血分离的CD4^+T细胞,经药物分别处理24h后,LCD+DXM组的激素敏感型和抵抗型的SLE CD4^+T细胞的总凋亡率均明显高于DXM组和空白组(均为P﹤0.01);DXM组的敏感型CD4^+T细胞总凋亡率明显高于空白组(P﹤0.01),而抵抗型的CD4^+T细胞总凋亡率在两组间无显著差异;LCD+DXM联合处理GC敏感型SLE患者外周血CD4^+T细胞24h后,CD4^+T细胞内标记GRα的FITC荧光抗体表达强度明显强于DXM组(P﹤0.05),但弱于空白组的荧光强度(P﹤0.05);LCD+DXM联合处理GC抵抗型SLE患者外周血CD4^+T细胞24h后,CD4^+T细胞内标记GRα的FITC荧光抗体表达强度明显强于DXM组和空白组的荧光强度(P﹤0.01和P﹤0.05);DXM组的GC敏感型和抵抗型SLE患者外周血CD4^+T细胞的标记GRα的FITC荧光抗体表达强度均低于空白组(均为P﹤0.01),GC抵抗型SLE患者外周血CD4^+T细胞内标记GRα的FITC荧光抗体表达低于GC敏感型SLE患者的CD4^+T细胞(P﹤0.01)。[结论]狼疮定能增强DXM诱导激素敏感型和抵抗型的SLE CD4^+T细胞凋亡,其作用可能与狼疮定拮抗DXM对激素敏感型和激素抵抗型SLE患者CD4^+T细胞胞浆内GRα表达的抑制作用相关;激素抵抗型SLE患者CD4^+T细胞胞浆内GRα表达低于�[Objeetive] To clarify the effect of Langchuangding（LCD） on the apoptosis and expressions of GRct of CD4cr cells in different glucocorticoid（GC） sensitive systemic lupus erythematosus（SLE）. [Methods] Screening GC resistant and susceptible SLE patients who consistent with the selected standard to collect the peripheral blood and sort CD4＋T cells immediately. Cultured CD4＋T cells were randomly divided into three groups： blank group（adding blank serum）, dexamethasone group（adding blank serum plus 10 μmol/ml dexamethasone（DXM）; traditional Chinese medicine combined western medicine group（CW group, adding LCD medicated serum 2.5ml plus 10 μmol/ml DXM）. After treated for 24h, CD4＋T cells were collected. The apoptosis of CD4＋T cells was detected by flow cytometry and the expression of GRct in CD4 ＋T cells was detected by immunofiuorescence technique, respectively. [Results] Of the 58 cases of SLE patients consisting with the selected standard, 25 cases of were glucocorticoid resistant and 33 cases were GC-susceptible. For CD4＋T cells of GC-susceptible SLE, the total apoptosis rates in CW group were significantly higher than those in DXM group and blank group（P〈0.01, respectively）, and there were significant differences between DXM group and blank group（P〈0.01）. For CD4＋T cells of glucocorticoid resistant SLE, the total apoptosis rates in CW group were significantly higher than those in DXM group and blank： group（P〈0.01, respectively）, but there were no significant differences between DXM group and blank group. Fluorescence intensities of FIT（： labeled GRct in GC-susceptible SLE CD4＋T cells of LCD＋DXM group were stronger than those of DXM group, but weaker than blank group（P〈0.05）. FITC fluorescence intensities in GC-resistant SLE CD4＋T cells of LCD＋DXM group were stronger than those of DXM and blank group（P〈0.01 and P〈0.05）. FITC fluorescence intensities in GC-susceptible and-resistant SLE CD4＋T ceils in DXM group were lower than

关 键 词：系统性红斑狼疮 狼疮定 糖皮质激素受体 敏感型 抵抗型 细胞凋亡 CD4+T 

分 类 号：R593[医药卫生—内科学]