Mas基因沉默对血管紧张素(17)拮抗血管紧张素Ⅱ诱导的肾间质成纤维细胞活化的影响

《Mas基因沉默对血管紧张素(17)拮抗血管紧张素Ⅱ诱导的肾间质成纤维细胞活化的影响》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、-interstitialfibroblastactivation,weuseMasSiRNAthatselectedalreadytotransienttransfecttheNRK-49F.Afterculturedfor48h,cellsisdividedinto6groups:①normalcontrolgroup(NG):thecellsaremaintainedintheDMEM-F12mediumsupplementedwith10%FBSand1%penicillin-streptomycin.②AngⅡgroup:thecellsaremaintainedintheDMEM-

2、F12mediumsupplementedwith10%FBSandtheangiotensinⅡthatthefinalconcentrationwas10-6mol/L.③Ang-(1-7)group:thecellsaremaintainedintheDMEM-F12mediumsupplementedwith10%FBSandtheangiotensin-(1-7)thatthefinalconcentrationwas10-5mol/L.④AngⅡ+Ang-(1-7)group:thecellsaremaintainedintheDMEM-F12mediumsupplementedw

3、ith10%FBS,10-6mol/LangiotensinⅡand10-5mol/Langiotensin-(1-7).⑤AngⅡ+Ang-(1-7)+negativeSiRNAgroup(A+SiRNA-CG):AfterbeentransienttransfectedbythenegativeSiRNAfor48h,thecellsaremaintainedintheDMEM-F12mediumsupplementedwith10%FBS,10-6mol/LangiotensinⅡand10-5mol/Langiotensin-(1-7).⑥AngⅡ+Ang-(1-7)+MasSiRNA

4、(A+MasSiRNA):AfterbeentransienttransfectedbytheMasSiRNASiRNAfor48h,thecellsaremaintainedintheDMEM-F12mediumsupplementedwith10%FBS,10-6mol/LangiotensinⅡand10-5mol/Langiotensin-(1-7).Thecellsaboveareculturedfor72h.Thentheexpressionofα-smoothmuscleactin(α-SMA)isdetectedbyimmunocytochemistrymethod.Theco

5、ntentofCollagenⅠ(ColⅠ)inthe6----culturedsupernatantismeasuredbyELISA.Results:①theresultsofimmunocytochemistry:AfterculturedwithAngⅡandAng-(1-7)for72h,thecellsofMasSiRNAtransfectiongroupcouldseehypertrophyandtheratioofcytoplasmisincreasing.Thereareaamountofboundaryclearbrowngranulesinthecytoplasmandi

6、ssimilartothecellsofAngⅡgroup.Andcomparedwiththenon-transfectiongroupandnegativeSiRNAtransfectiongroup,theexpressionofα-SMAincreasedsignificantlyinthecellafteranalysisofdata(P<0.05).TherearenosignificantdifferencebetweenAngⅡ+Ang-(1-7)groupandAngⅡ+Ang-(1-7)+negativeSiRNAgroup(P>0.05).Incontrolgroup,t

7、hereisbasicexpressionsofα-SMA.②ELISA:AfterculturedwithAngⅡandAng-(1-7)for72h,thesecretionofColⅠisrisedsignificantlyintheculturedsupernatantofMasSiRNAtransfectiongroupthanthenon-transfectiongroupandneg