P38丝裂原活化蛋白激酶短发夹RNA对H9C-2心肌细胞基因表达的影响  

Suppression of P38 MAPK expression in H9C-2 myocardial cells by short hair RNA

作　　者：梁远红[1] 周燕[2] 刘烈[1] 陈东骊[1] 林纯莹[1] 陈泗林[1] 林曙光[1] 

机构地区：[1]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),广东广州510080 [2]武汉大学人民医院急诊科,湖北武汉430060

出　　处：《中国现代医学杂志》2012年第32期22-25,共4页China Journal of Modern Medicine

基　　金：中国博士后科学基金(No:20060400780);广东省医学科研基金(No:A2007022);广东省自然科学基金(No:7001120)

摘　　要：目的探讨P38丝裂原活化蛋白激酶(P38MAPK)的短发夹环RNA(shRNA)质粒对H9C-2心肌细胞增殖变化的影响并探讨其发生机制。方法构建3种P38MAPK shRNA质粒并测序鉴定,将其转染入H9C-2心肌细胞中,应用RT-PCR和Western blotting检测心脏细胞P38MAPK mRNA和蛋白的表达。结果P38MAPK shRNA质粒分布在心肌细胞的胞浆及细胞核中,与心肌细胞组比较,AngⅡ组和shRNA阴性组的P38MAPK mRNA和蛋白水平显著升高(P<0.01,P<0.01);与AngⅡ组比较,shRNA1、shRNA2和shRNA3组的P38MAPK mRNA和蛋白水平明显降低(P<0.05,P<0.05,P<0.01)。结论 P38MAPK shRNA质粒成功转染心肌细胞,P38MAPK shRNA3质粒能最有效地抑制心肌细胞P38MAPK的表达。【Objective】 To evaluate the effects of P38 mitogen-activated protein kinase（MAPK） short hair RNA（shRNA） plasmid delivery by Lipofectamin on mRNA and protein expression in H9C-2 myocardial cells.【Methods】 Three P38MAPK shRNA sequences were designed and synthesized,and subcloned into GPU6/ GFP/Neo plasmid.After restriction enzyme digestion and sequencing,the selected positive recombinant plasmid was transfected into H9C-2 myocardial cells.The mRNA and protein expressions of P38MAPK in H9C-2 myocardial cells were analyzed by RT-PCR and Western blotting analysis respectively.【Results】 It showed that the P38MAPK shRNA GPU6/GFP/Neo plasmid was taken up into H9C-2 myocardial cells.Compared with those in the H9C-2 myocardial cell group,the P38MAPK mRNA and protein levels in Ang Ⅱ group and plasmid control group significantly increased（P 0.01,P 0.01;respectively）.Compared with those in the Ang Ⅱ group,the P38MAPK mRNA and protein levels in shRNA1,shRNA2 and shRNA3 group significantly decreased（P 0.05,P 0.05,P 0.01;respectively）.【Conclusion】 P38MAPK shRNA plasmid could be transfected into cultured myocardial cells triumphally.P38MAPK shRNA3 is the most effective shRNA which inhibited the level of P38MAPK mRNA and protein expression.

关 键 词：P38丝裂原活化蛋白激酶 短发夹RNA 心肌细胞 

分 类 号：Q786[生物学—分子生物学]