rho激酶抑制剂y-27632对c3h10t12细胞增殖与成脂分化的作用

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1、ISSN1007-7626中国生物化学与分子生物学报http://cjbmb．bjmu．edu．cn2013年7月CN11-3870/QChineseJournalofBiochemistryandMolecularBiology29(7):661～666Rho激酶抑制剂Y-27632对C3H10T1/2细胞增殖与成脂分化的作用1)，2)2)2)*王新鲁，姬云涛，屈长青1)2)(安徽大学生命科学学院，安徽合肥230601;抗衰老中草药安徽省工程技术研究中心，安徽阜阳236037)摘要探究Rho激酶抑制剂Y-27632对间充质干细胞(m

2、esenchymalstemcells，MSCs)C3H10T1/2增殖和成脂分化的影响．实验分为对照组、成脂诱导组和Y-27632处理组(Y-27632+成脂诱导)．利用MTT检测细胞增殖情况，油红O染色，异丙醇萃取法检测细胞成脂分化情况，半定量RT-PCR检测过氧化物酶体增殖物激活受体γ(peroxisomeproliferatoractiviatedreceptorγ，PPARγ)和CCAAT增强子结合蛋白α(CCAATenhancerbindingproteinα，C/EBPα)基因表达．结果表明，Y-27632能够显著抑制C

3、3H10T1/2细胞的增殖(P＜0.05)，并呈一定的浓度依赖性;高浓度Y-27632对C3H10T1/2细胞成脂分化具有显著抑制作用(P＜0.05);半定量RT-PCR结果显示，成脂诱导处理组PPARγ和C/EBPα表达量在第3d、5d和7d显著低于成脂诱导组(P＜0.05)．综上所述，Y-27632能够抑制C3H10T1/2细胞增殖与成脂分化．关键词Rho激酶抑制剂Y-27632;间充质干细胞;增殖;成脂分化中图分类号Q25EffectofRhoKinasesInhibitorY-27632onProliferationandAd

4、ipogenicDifferentiationofMesenchymalStemCellC3H10T1/21)，2)2)2)*WANGXin-Lu，JIYun-Tao，QUChang-Qing1)(CollegeofLifeSciences，AnhuiUniversity，Hefei230601，China;2)EngineeringTechnologyResearchCenterofAnti-agingChineseHerbalMedicine，Fuyang236037，Anhui，China)AbstractToinvestiga

5、tetheeffectofRhokinasesinhibitorY-27632ontheproliferationandadipogenicdifferentiationofmesenchymalstemcells，theC3H10T1/2cellswereculturedinDMEMmediumwith10%fetalbovineserumat37℃in5%CO2incubatoranddividedintothecontrol，adipogenicandY-27632groups．Thecellproliferationwasde

6、tectedbyMTT;theadipogenicdifferentiationwascharacterizedbyoilredOstaining．TheC3H10T1/2celldifferentiationintoadipocyteswasalsoexaminedbythegeneexpressionofPPARγandC/EBPαbyRT-PCR．TheresultsshowedthatY-27632inhibitedtheproliferationofC3H10T1/2cellinadose-dependentmanner(P

7、＜0.05)．HighconcentrationsofY-27632inhibitedadipogenicdifferentiationofC3H10T1/2cell(P＜0.05)．TheRT-PCRassaysshowedthatthePPARγandC/EBPαexpressionwereinhibitedbyY-27632atday3，5and7duringdifferentiationascomparedtothecontrol(P＜0.05)．Insummary，Y-27632isabletoinhibittheproli

8、ferationandadipogenicdifferentiationofC3H10T1/2cell．KeywordsRhokinasesinhibitorY-27632;mesenchymalstemcell;pro