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ID:26890353
大小:555.50 KB
页数:48页
时间:2018-11-29
《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
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