STAT5基因沉默对支气管哮喘小鼠T细胞增殖的影响  

Effect of STAT5 gene silencing on the proliferation of T lymphocytes in a mouse model of asthma

作　　者：邱晨[1] 张婷[1] 齐晖[2] 彭文科[1] 史菲[1] 

机构地区：[1]暨南大学第二临床医学院呼吸内科,深圳518020 [2]暨南大学第二临床医学院中心实验室,深圳518020

出　　处：《中华结核和呼吸杂志》2012年第1期50-54,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：深圳市科技局重点课题（200801014）

摘　　要：目的探讨STATSa／b基因沉默对支气管哮喘（简称哮喘）小鼠T细胞增殖功能的影响。方法将20只雌性BALB／c小鼠按随机数字法分为正常组和哮喘组，哮喘组按处理因素不同分别分为空白对照组、STATSa沉默组、STAT5b沉默组及错义链对照组，采用免疫磁珠法筛选出正常及哮喘模型小鼠的脾脏T细胞，利用RNA干扰（RNAi）沉默T细胞内STATSa／b基因。荧光定量PCR检测STATSa／b基因沉默前后STAT5a／b的mRNA表达变化；蛋白免疫印迹法检测STATSa／b基因沉默前后的蛋白含量的变化；CCK-8检测T细胞RNA干扰前后增殖率的变化；流式技术检测T细胞RNA干扰前后细胞增殖周期及早期凋亡率的变化。结果（1）哮喘小鼠气道病理结果显示明显炎症改变。（2）哮喘组小鼠脾脏淋巴细胞STAT5a／bmRNA表达量较正常组小鼠明显增高（r值分别为11．71、33．04，均P〈0．05）。（3）与空白对照组比较：特异性siRNA转染哮喘小鼠T细胞后，STATSa和STAT5b沉默组的mRNA表达明显降低（T值分别为35．014、14．553，均P〈0．01）；STAT5a和STAT5b沉默组蛋白表达也明显降低（r值分别为-10．958、-14．706，均P〈0．01）；STAT5a和STATSb沉默组T细胞增殖率明显降低（r值分别为8．692、10．540，均P〈0．01）；STATSa和STATSb沉默组增殖期T细胞均明显降低（T值分别为-6．975、-5．567，均P〈0．05）；STATSa和STATSb沉默组T细胞凋亡率明显升高（r值分别为-6．404、-6．038，均P〈0．05）。结论RNA干扰能特异性降低哮喘小鼠STATSa／b表达，抑制T细胞的增殖并促进T细胞凋亡。Objective To study the effect of silencing STAT5 gene on the proliferation of T lymphoeytes in a mouse model of asthma. Methods Spleen T-lymphoeytes of normal and asthmatic mice were selected by immunomagnetie beads, and the STAT5a/b genes of these T-lymphoeytes were silenced by siRNA. The mRNA of STAT5a/b was detected by fluorescence quantitative PCR, and the protein by Western blot. The proliferation rates of T-lymphoeytes was evaluated by CCK-8, and the cell cycle and the cell apoptosis was measured by flow eytometry. Results （ 1 ） Significant inflammatory cell infiltration was present in asthmatic mouse airways, as compared to the normal control mice. （2） The STAT5a/b mRNA expression of the asthmatic group was significantly higher compared with the normal control group. （3） Compared to the blank control group, the mRNA expression of STAT5a/b was reduced in the siRNA-specific intervention（ T： 35.014 vs 14. 553, P 〈0. 01 ）. The protein expression of STAT5a/b was also reduced after siRNA-specific intervention（ T： - 10. 958 vs - 14. 706, P 〈 0.01 ）. The proliferation rates of T lymphocytes was reduced after siRNA-specific intervention （ T： 8. 692 vs 10. 540, P 〈 0. 01 ）, and the number of T-lymphocytes in proliferative phase was all significantly reduced after siRNA-specific intervention （T： 6. 975 vs - 5. 567, P 〈 0. 05 ）. The apoptosis rates of T lymphocytes were significantly increased after siRNA-specific intervention （ T： - 6. 404 vs - 6. 038, P 〈 0. 05 ） . Conclusion RNA interference specifically reduced the expression of STAT5a/b, inhibited the proliferation of T lymphocytes, and promoted the apoptosis of T lymphocytes in a mouse model of asthma.

关 键 词：哮喘 T淋巴细胞 STAT5转录因子 

分 类 号：R562.25[医药卫生—呼吸系统]