CTGF反义寡核苷酸对AngⅡ诱导的人近曲肾小管上皮细胞转分化的影响.doc

CTGF反义寡核苷酸对AngⅡ诱导的人近曲肾小管上皮细胞转分化的影响                                      作者:陈珑,刘必成,马坤岭,刘宏,罗冬冬目的:探讨结缔组织生长因子(CTGF)反义寡核苷酸对血管紧张素Ⅱ(AngⅡ)诱导的肾小管细胞转分化的可能影响。方法:采用体外培养的人近曲肾小管上皮细胞株(HK2),分别观察CTGF反义寡核苷酸和AngⅡ干预细胞对细胞CTGFmRNA和蛋白质的表达、细胞内超微结构及细胞α平滑肌肌动蛋白(αSMA)表达的影响。结果:AngⅡ干预HK2细胞48h后,细胞CTGFmRNA和蛋白的表达显著增高(P<);AngⅡ干预96h后,细胞超微结构显示出间质细胞样结构;这些作用都可被CTGF反义寡核苷酸显著抑制。AngⅡ可以呈时间依赖性地刺激HK2细胞表达αSMA,这些作用可以被CTGF反义寡核苷酸显著抑制(P<)。对照组错义寡核苷酸无上述作用。结论:CTGF反义寡核苷酸对AngⅡ诱导HK2细胞转分化具有显著的抑制作用,提示CTGF可能介导了AngⅡ诱导的肾小管上皮细胞转分化。[关键词]人近曲肾小管上皮细胞;血管紧张素Ⅱ;结缔组织生长因子;细胞转分化 Abstract:Objective Toinvestigatetheeffectofconnectivetissuegrowthfactorantisenseoligonucleotide(CTGFAS)onAngⅡ TheinfluenceofCTGFASonAngⅡinducedHK2cellsCTGFmRNAandproteinexpressionwereobservedbyRTPCRandconfocalmicroscopyrespectively. TheeffectofCTGFASonAngⅡinducedcellularultrastructurewereobservedbytransmissiveelectronicmicroscope(TEM). Theexpressionofαsmoothaction(αSMA)wereobservedbyimmunocytochemistry. Thecontrolgroupwastreatedwithscrambledoligonucleotide(SC).Results ItwasshownthatAngⅡsignficantlyinducedtheincreasingexpressionofCTGFmRNAandprotein(P<,respectively).IfthecellswerestimulatedwithAngⅡfor96h,thecellularultrastructureshowedmesenchymalfeatures. AndtheseeffectscouldbepartiallyattenuatedbyCTGFAS. AngⅡsignificantlystimulatedtheexpressionofαSMAinatimedependentmanner,whichcouldbemarkedlyattenuatedbythetreatmentwit