慢病毒介导的p38丝裂原活化蛋白激酶RNA干扰对心肌成纤维细胞结缔组织生长因子的影响  

Effects of ientiviral-mediated p38 mitogen-activated protein kinase RNA interference on connective tissue growth factor expression in myocardial fibroblasts

作　　者：周燕[1] 魏捷[1] 梁远红[3] 陈静[1] 唐其柱[2] 

机构地区：[1]武汉大学人民医院急诊科,430060 [2]武汉大学人民医院心内科,430060 [3]广东省心血管病研究所广东省医学科学院广东省人民医院

出　　处：《中华实验外科杂志》2013年第5期1002-1004,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：湖北省自然科学基金资助项目（2010CDB06806）;广东省医学科研基金资助项目（A2007022）

摘　　要：目的观察慢病毒介导的p38丝裂原活化蛋白激酶（p38MAPK）的短发夹环RNA（shRNA）对心肌成纤维细胞结缔组织生长因子（CTGF）的影响并探讨其机制。方法构建慢病毒p38MAPKshRNA（PGLV—shRNA）并测序鉴定，观察其对转化生长斟子-β（TGF—β）诱导心肌成纤维细胞的影响，检测p38MAPK、CTGFmRNA和CTGF、α-平滑肌肌动蛋白（α—SMA）蛋白的表达。结果TGF—β4nmol／L刺激24h使心肌成纤维细胞p38MAPKmRNA、CTGFmRNA、CTGF蛋白和α—SMA蛋白明显升高（P〈0．01）。PGLV—shRNA明显减少TGF．13诱导的p38MAPKmRNA、CTGFmRNA、CTGF蛋白和α-SMA蛋白表达（0．252±0．041比0．652±0．089，P〈0．01；0．418±0．071比0．838±0．099，P〈0．01；0．418±0．076比0．991±0．117，P〈0．01；0．465±0．069比0．875±0．100，P〈0．05）。结论TGF—经p38MAPK信号通路诱导心肌成纤维细胞CTGF表达，PGLV—shRNA能有效抑制TGF—α诱导的心肌细胞纤维化。Objective To explore the effects of p38 mitogen-activated protein kinase （MAPK） short-hair llNA （shlRNA） delivered by lentiviral vectors （PGLV） on connective tissue growth factor （CTGF） expression in myocardial fibroblasts. Methods The PGLV-shtlNA was transfected into myocardial fihroblasts to to explore the role of p38MAPK pathway activation in transforming growth factor （TGF）-β-mediated myoeardial fibrosis. The mtlNA expression levels of p38MAPK and CTGF were detected by using reverse transeriptase-polymerase chain reaction （ RT-PCR）, and the protein expression levels of CTGF and α-smooth muscle actin （α-SMA） by using Western blotting. Results TGF-13 stimulation inereased p38MAPK and CTGF retINA, and CTGF and α-SMA protein expression in myocardial fibroblasts （P 〈 0. 01）. As compared with those in TGF-β group, PGLV-shRNA transfeetion decreased p38MAPK and CTGF retINA, and CTGF and β-SMA protein expression in myocardial fibroblasts [ （0. 252 ± 0. 041 vs. 0.652.±0.089,P〈0.01; 0.418 ±0.071 vs. 0.838 ±0.099,P〈0.01; 0.418 ±0.076 vs. 0.991 ± 0. 117,P〈0. 1; 0.465 ±0.069 vs. 0.875 ±0. 100,P〈0.05） ]. Conclusion TGF-β directly upregulates CTGF expression through p38MAPK pathway and gene silence of p38MAPK may protect TGF-β-mediated fibrosis in cardiac myoeytes.

关 键 词：P38丝裂原活化蛋白激酶 慢病毒 结缔组织生长因子 RNA干扰 

分 类 号：R542[医药卫生—心血管疾病]