锌转运蛋白1 siRNA转染骨髓间充质干细胞对肿瘤坏死因子α信号成脂通路的影响  

Effect of bone marrow mesenchymal stem cells transfected by Zrt/Irt-like protein 1 on adipogenic differentiation via tumor necrosis factor alpha signaling pathway

作　　者：章晓云[1] 黎金焕 袁振中[1] 陈跃平[1] 夏天[2] 卓映宏 冯洋[2] 蓝佼 董盼锋[1] 赵斌[2] 

机构地区：[1]广西中医药大学附属瑞康医院骨科,广西壮族自治区南宁市530011 [2]广西中医药大学,广西壮族自治区南宁市530011

出　　处：《中国组织工程研究》2017年第13期1986-1991,共6页Chinese Journal of Tissue Engineering Research

基　　金：广西自然科学基金课题(2015GXNSFAA139136);广西高校青年教师基础能力提升项目(KY2016YB204);广西卫生厅重点课题(S201419-05);广西中医药大学校级科研项目(2015LX027);2016年研究生教育创新计划资助项目立项课题(YJS201650);2014年广西中医药大学校级培育学科-骨外科学建设项目[J13167(7)]~~

摘　　要：背景:课题组前期研究已经证实乙醇能促进骨髓间充质干细胞成脂分化,上调肿瘤坏死因子α成脂通路蛋白PPARγ,aP 2的表达。锌转运蛋白1(Zrt/Irt-like Protein 1,ZIP1)作为ZIP蛋白家族的成员与骨代谢和成骨分化有密切的关系。目的:观察ZIP1 siR NA转染的骨髓间充质干细胞对肿瘤坏死因子α信号成脂通路的影响。方法:分离培养兔骨髓间充质干细胞,使用0.03,0.09,0.15,0.21 mol/L乙醇进行培养,再使用ZIP1 siR NA转染骨髓间充质干细胞,同时对兔骨髓间充质干细胞进行ZIP1转染培养。结果与结论:(1)aP 2,PPARγ蛋白的表达:不同浓度乙醇培养aP 2,PPARγ蛋白表达均较明显升高(P<0.05);除0.03 mol/L乙醇组之外,其他浓度乙醇组三酰甘油水平明显升高(P<0.05);沉默ZIP1基因转染乙醇培养的骨髓间充质干细胞后,各乙醇浓度组a P2,PPARγ蛋白的表达量及三酰甘油水平均明显升高(P<0.05),而骨髓间充质干细胞经ZIP1表达载体转染后,a P2,PPARγ蛋白表达明显降低(P<0.05);(2)结果表明:沉默ZIP1表达能激活骨髓间充质干细胞的肿瘤坏死因子α信号成脂通路的表达,从而发挥促进细胞的成脂分化能力。BACKGROUND：Previous studies have confirmed that ethanol can promote adipogenic differentiation of bone marrow mesenchymal stem cells （BMSCs）, and up-regulate the expression of PPARγ and aP2 in the tumor necrosis factor α （TNF-α） signaling pathway. As a member of the ZIP protein family, Zrt/Irt-like protein 1 （ZIP1） is closely related to bone metabolism and osteogenic differentiation.OBJECTIVE：To study the effect of BMSCs transfected by ZIP1 on TNF-α signaling pathway in the process of adipogenic differentiation.METHODS：The BMSCs from rabbits were isolated and cultured under different concentrations of alcohol （0.03, 0.09,0.15, 0.21 mol/L）, followed by transfection by ZIP1 siRNA and ZIP1 expression vector.RESULTS AND CONCLUSION：After culture in alcohol, the expression levels of aP2 and PPARγ proteins were both significantly increased （P 〈 0.05）, and the level of triglyceride was increased in all alcohol groups except for 0.03 mol/L alcohol group （P 〈 0.05）. After siRNA transfection, the expression levels of aP2 and PPARγ as well as the level of triglyceride were increased significantly in all the alcohol groups （P 〈 0.05）; however, ZIP1 transfection decreased the expression levels of aP2 and PPARγ proteins （P 〈 0.05）. To conclude, ZIP1 siRNA could promote the adipogenic differentiation of BMSCs through the activation of TNF-α signaling pathway.

关 键 词：干细胞 股骨头坏死 组织工程 骨髓干细胞 骨髓间充质干细胞 锌转运蛋白 成脂通路 三酰甘油 过氧化物酶体增殖物激活受体γ 脂肪细胞型脂肪酸结合蛋白 蛋白免疫印迹 乙醇 广西壮族自治区自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]