葛根素上调miR-155-3p降低内脏脂肪素诱导人脐静脉内皮细胞损伤  

Puerarin alleviates HUVEC injury induced by visfatin through up-regulating miR-155-3p

作　　者：周凤华[1] 黄志勇[2] 张蕾[1] 孙学刚[1] 贾钰华[1] 

机构地区：[1]南方医科大学中医药学院,广州510515 [2]南方医科大学第三附属医院眼耳鼻咽喉科,广州510630

出　　处：《中国实验动物学报》2016年第5期465-469,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(No.81373574;81403339);广东省自然科学基金博士启动项目(No.2014A030310150);广东省中医药管理局科研项目(No.20141186);广东省教育部产学研结合项目(2012B091100157);南方医科大学科研启动计划(No.PY2013N014)

摘　　要：目的研究葛根素对内脏脂肪素(visfatin)诱导人脐静脉内皮细胞(HUVEC)炎症因子及miR-155-3p表达的影响。方法采用visfatin干预HUVEC建立细胞损伤模型,不同浓度(0.5,1.0,2.0 g/L葛根素分别干预细胞24 h,采用MTT法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,ELISA法检测细胞内CRP和NF-κB的水平,RT-PCR法检测细胞内miR-155-3p水平,western blotting测髓样分化因子(myeloid differentiation factor 88,My D88)蛋白水平。结果 Visfatin能显著抑制HUVEC增殖,并且诱导其凋亡,与对照组相比差异有显著性(P<0.01),葛根素中、高浓度组可明显抑制visfatin对HUVEC细胞损伤作用;此外,葛根素中、高浓度组能显著降低细胞上清CRP和NF-κB水平,增加细胞内miR-155-3p基因表达,与模型组相比差异有显著性(P<0.01)。增加miR-155-3p水平可显著降低下游My D88蛋白表达,抑制HUVEC分泌CRP与NF-κB,与模型组相比差异有显著性(P<0.05)。结论葛根素能显著减轻visfatin诱导HUVEC炎症损伤效应,其机制可能与葛根素上调miR-155-3p水平抑制靶基因My D88蛋白表达,从而降低细胞内CRP与NF-κB水平有关。Object To study the effect of puerarin on the expression of inflammatory factors and miR-155-3p in human umbilical vein endothelial cells（ HUVEC） induced by visfatin. Methods The HUVEC cell injury model was established with visfatin. Cell proliferation was measured by MTT assay. Cell apoptosis was detected by flow cytometry. The level of CRP and NF-κB was detected by ELISA,and the expression of miR-155-3p was detected by RT-PCR. The expression of myeloid differentiation factor 88（ My D88） was identified by western blotting. Results Visfatin induced cell proliferation and inhibited apoptosis in HUVEC,meanwhile the expressions of both CRP and NF-κB were significantly increased,compared with that of the control group（ P〈0. 01）. Puerarin at moderate and high concentrations obviously reduced the HUVEC injury induced by visfatin,mainly through down-regulating the expression of CRP and NF-κB,as well as up-regulating the level of miR-155-3p in the HUVEC. MiR-155-3p mimic markedly decreased the level of My D88,CRP and NF-κB in the HUVEC induced by visfatin（ P〈0. 05）. Conclusions Pueprarin obviously alleviates HUVEC injury inducedby visfatin,probably related to down-regulating the level of MyD 88,CRP,NF-κB,and up-regulating the expression of miR-155-3p in HUVEC.

关 键 词：人脐静脉内皮细胞 内脏脂肪素 葛根素 miR-155-3p 

分 类 号：Q95-33[生物学—动物学]