中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2012年
1期
50-54
,共5页
邱晨%张婷%齐晖%彭文科%史菲
邱晨%張婷%齊暉%彭文科%史菲
구신%장정%제휘%팽문과%사비
哮喘%T淋巴细胞%STAT5转录因子
哮喘%T淋巴細胞%STAT5轉錄因子
효천%T림파세포%STAT5전록인자
Asthma%T-lymphocytes%STAT5 transcription factor
目的 探讨STAT5a/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、STAT5a沉默组、STAF5b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STAT5a/b基因.荧光定量PCR检测STAT5a/b基因沉默前后STAT5a/b的mRNA表达变化；蛋白免疫印迹法检测STAT5a/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/b mRNA表达量较正常组小鼠明显增高(T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,STAT5a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(T值分别为- 10.958、- 14.706,均P＜0.01)；STAT5a和STAT5b沉默组T细胞增殖率明显降低(T值分别为8.692、10.540,均P＜0.01)；STAT5a和STAT5b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)；STAT5a和STAT5b沉默组T细胞凋亡率明显升高(T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STAT5 a/b表达,抑制T细胞的增殖并促进T细胞凋亡.
目的 探討STAT5a/b基因沉默對支氣管哮喘(簡稱哮喘)小鼠T細胞增殖功能的影響.方法 將20隻雌性BALB/c小鼠按隨機數字法分為正常組和哮喘組,哮喘組按處理因素不同分彆分為空白對照組、STAT5a沉默組、STAF5b沉默組及錯義鏈對照組,採用免疫磁珠法篩選齣正常及哮喘模型小鼠的脾髒T細胞,利用RNA榦擾(RNAi)沉默T細胞內STAT5a/b基因.熒光定量PCR檢測STAT5a/b基因沉默前後STAT5a/b的mRNA錶達變化；蛋白免疫印跡法檢測STAT5a/b基因沉默前後的蛋白含量的變化；CCK-8檢測T細胞RNA榦擾前後增殖率的變化；流式技術檢測T細胞RNA榦擾前後細胞增殖週期及早期凋亡率的變化.結果 (1)哮喘小鼠氣道病理結果顯示明顯炎癥改變.(2)哮喘組小鼠脾髒淋巴細胞STAT5a/b mRNA錶達量較正常組小鼠明顯增高(T值分彆為11.71、33.04,均P＜0.05).(3)與空白對照組比較:特異性siRNA轉染哮喘小鼠T細胞後,STAT5a和STAT5b沉默組的mRNA錶達明顯降低(T值分彆為35.014、14.553,均P＜0.01)；STAT5a和STAT5b沉默組蛋白錶達也明顯降低(T值分彆為- 10.958、- 14.706,均P＜0.01)；STAT5a和STAT5b沉默組T細胞增殖率明顯降低(T值分彆為8.692、10.540,均P＜0.01)；STAT5a和STAT5b沉默組增殖期T細胞均明顯降低(T值分彆為-6.975、-5.567,均P＜0.05)；STAT5a和STAT5b沉默組T細胞凋亡率明顯升高(T值分彆為-6.404、-6.038,均P＜0.05).結論 RNA榦擾能特異性降低哮喘小鼠STAT5 a/b錶達,抑製T細胞的增殖併促進T細胞凋亡.
목적 탐토STAT5a/b기인침묵대지기관효천(간칭효천)소서T세포증식공능적영향.방법 장20지자성BALB/c소서안수궤수자법분위정상조화효천조,효천조안처리인소불동분별분위공백대조조、STAT5a침묵조、STAF5b침묵조급착의련대조조,채용면역자주법사선출정상급효천모형소서적비장T세포,이용RNA간우(RNAi)침묵T세포내STAT5a/b기인.형광정량PCR검측STAT5a/b기인침묵전후STAT5a/b적mRNA표체변화；단백면역인적법검측STAT5a/b기인침묵전후적단백함량적변화；CCK-8검측T세포RNA간우전후증식솔적변화；류식기술검측T세포RNA간우전후세포증식주기급조기조망솔적변화.결과 (1)효천소서기도병리결과현시명현염증개변.(2)효천조소서비장림파세포STAT5a/b mRNA표체량교정상조소서명현증고(T치분별위11.71、33.04,균P＜0.05).(3)여공백대조조비교:특이성siRNA전염효천소서T세포후,STAT5a화STAT5b침묵조적mRNA표체명현강저(T치분별위35.014、14.553,균P＜0.01)；STAT5a화STAT5b침묵조단백표체야명현강저(T치분별위- 10.958、- 14.706,균P＜0.01)；STAT5a화STAT5b침묵조T세포증식솔명현강저(T치분별위8.692、10.540,균P＜0.01)；STAT5a화STAT5b침묵조증식기T세포균명현강저(T치분별위-6.975、-5.567,균P＜0.05)；STAT5a화STAT5b침묵조T세포조망솔명현승고(T치분별위-6.404、-6.038,균P＜0.05).결론 RNA간우능특이성강저효천소서STAT5 a/b표체,억제T세포적증식병촉진T세포조망.
Objective To study the effect of silencing STAT5 gene on the proliferation of T lymphocytes in a mouse model of asthma.Methods Spleen T-lymphocytes of normal and asthmatic mice were selected by immunomagnetic beads,and the STAT5a/b genes of these T-lymphocytes 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-lymphocytes was evaluated by CCK-8,and the cell cycle and the cell apoptosis was measured by flow cytometry.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.
