沉默ERRα对过表达DKK1、Sost重组腺病毒载体转染的MG63细胞影响研究  

Effect of Silencing ERRa in DKK1,Sost Overexpressing Recombinant Adenovirus Vector Infected on MG63 Cells

作　　者：刘少津 乔荣勤[2] 万雷[2] 黄宏兴[2] 魏合伟[2] LIU Shao-jin;WAN Lei;HUANG Hong-xing;WEI He-wei;QIAO Rong-qin(Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510405;The Affiliated Orthopedics and Trauma Hospital, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510240)

机构地区：[1]广州中医药大学,广东广州510240 [2]广州中医药大学附属骨伤科医院,广东广州510405

出　　处：《按摩与康复医学》2019年第9期36-39,共4页Chinese Manipulation and Rehabilitation Medicine

基　　金：国家自然科学基金资助项目;编号:81302991;81373653;广东省科技计划项目;编号:2016A020216024

摘　　要：目的:探讨雌激素相关受体α(ERRα)对Wnt信号通路抑制因子Dickkopf(DKK)1、骨硬化蛋白Sclerostin(Sost)过表达腺病毒载体转染的MG63细胞和相关蛋白Lrp5、CTGF、FGF2、OPN的影响作用。方法:将培养好的MG63细胞经过表达DKK1、Sost转染后分为空白对照组、过表达DKK1组、过表达Sost组、过表达(DKK1+Sost)组,根据组别不同用包装好的沉默ERRα干预过表达DKK1、Sost重组腺病毒载体转染后的MG63细胞,采用四甲基偶氮唑蓝微量酶反应比色法(MTT)检测细胞增殖、考马斯亮蓝蛋白定量法测定ALP活性、流式分析法测定钙离子浓度,Western blot检测MG63细胞中Lrp5、CTGF、FGF2、OPN蛋白的表达量。结果:①与空白对照组比较,过表达DKK1、Sost、DKK1+Sost组、沉默ERRα干预空载腺病毒组的高细胞活性、ALP活性和Lrp5、CTGF、FGF2、OPN的表达量均降低,钙离子浓度均升高,差异均有统计学意义(P<0.05)。②与过表达DKK1、Sost、DKK1+Sost组比较,沉默ERRα干预过表达DKK1、Sost、DKK1+Sost组的细胞活性、ALP活性和Lrp5、CTGF、FGF2、OPN的表达量均降低,钙离子浓度均升高,差异均有统计学意义(P<0.05)。结论:沉默ERRα可降低过表达DKK1、Sost重组腺病毒转染后的MG63细胞增殖和ALP活性,升高钙离子浓度,对Lrp5、CTGF、FGF2、OPN蛋白表达有一定影响作用,尤其对DKK1、Sost同时过表达时的作用最为明显。Objective: To investigate the effect of the silencing of ERRa inhibitor on Dickkopfl(DKK1), sclerostin(Sost) reconmbinant adenovirus vector, the transfection MG63 cells and the Lrp5, CTGF, FGF2, OPN proteins expression. Methods: The cultured mature MG63 cells were divided into blank control group, DKK1 overexpression group, Sost overexpression group, and DKK1+Sost overexpression group. MG63 cells of different groups were infected by the corresponding of silence ERRa, after it divided into ERRa intervention empty adenovirus group, ERRa intervention overexpression DKK1 group, ERRa intervention overexpression Sost group, ERRa intervention overexpression DKK1 + Sost group, and then we observed the cell viability, alkaline phosphatase(ALP) activity, and Ca2+ concentration and the Lrp5, CTGF, FGF2, OPN proteins expression. Results:① Compared with the blank control group, the cells optical density(OD) value, ALP activity and Lrp5, CTGF, FGF2, OPN were significantly decreased, and Ca2+ concentration was significantly increased in overexpression Sost group, overexpression DKKl+Sost group, and ERRα intervention empty adenovirus group(P<0.05).②Compared with the overexpression DKK1, Sost, DKK1+Sost group, silencimg of ERRa interfered with overexpression of DKK1, Sost, and DKK1+Sost groups of the cells optical density(OD) value and ALP activity and the Lrp5, CTGF, FGF2, OPN were significantly decreased, and the Ca2+ concentration was significantly increased(P<0.05). Conclusion: The silencing of ERRa can decrease the overexpression of DKK1, Sost reconmbinant adenovirus vector infected on MG63 cell viability and ALP activity, increase Ca2+ concentration, and has certain effects on Lrp5, CTGF, FGF2, OPN related bone regulating proteins, and the effects of the overexpression of DKK1+Sost are strongest.

关 键 词：雌激素受体相关受体α(ERRα) DKK1 SOST 腺病毒 MG63细胞 过表达 转染 

分 类 号：R580[医药卫生—内分泌]