miR-194-3p调控模拟微重力条件下致成骨细胞功能异常的实验研究  

MiR-194-3p Regulation of Simulated Microgravity Induced�Dysfunction of Osteoblasts:An Experimental Study

作　　者：薛进冬 程路畅 王敏[1] 谭彩美 邓琪琪 朱海美 郭勇[1,2] 韩标 XUE Jindong;CHENG Luchang;WANG Min;TAN Caimei;DENG Qiqi;ZHU Haimei;GUO Yong;HAN Biao(Intelligent Medicine and Biotechnology School,Guilin Medical University,Guilin 541199,China;Key Laboratory of Medical Biotechnology and Translational Medicine,Guilin 541199,China)

机构地区：[1]桂林医学院智能医学与生物技术学院,桂林541199 [2]广西高校医药生物技术与转化医学重点实验室,桂林541199

出　　处：《医用生物力学》2024年第4期623-630,共8页Journal of Medical Biomechanics

基　　金：广西自然科学基金项目(2020GXNSFBA238006,桂科AD22080037);国家自然科学基金项目(12202113,32071309);广西硕士研究生创新项目(YCSW2022368)。

摘　　要：目的探讨模拟微重力环境下miR-194-3p对成骨细胞功能变化的调控作用,为极端力学环境下成骨细胞的力学响应机制提供理论基础。方法利用细胞旋转培养系统建立细胞水平微重力环境,分别采用MTT、RT-PCR、Western blot、荧光双染色法和茜素红染色法检测转染miR-194-3p抑制剂前后MC3T3-E1成骨细胞增殖、分化、凋亡和矿化的变化情况。结果模拟微重力环境下miR-194-3p表达上调,成骨细胞的增殖、分化和矿化均受到一定程度的抑制,同时促进了成骨细胞的凋亡。转染miR-194-3p抑制剂后,miR-194-3p表达显著下调,且能部分逆转由微重力导致的细胞成骨细胞增殖减弱、成骨分化标志物如ALP、OCN和COL-I基因和蛋白表达降低、骨矿化结节减少和成骨细胞凋亡数目增加的情况,说明miR-194-3p能有效改善微重力暴露下成骨细胞功能异常的情况。结论模拟微重力环境下miR-194-3p可能作为一种负调控因子,通过调控成骨细胞功能参与其力学响应的进程。Objective To investigate the role of miR-194-3p in regulating functional changes in osteoblasts in a simulated microgravity environment and to provide a theoretical foundation for understanding the mechanical response mechanisms of osteoblasts in extreme mechanical environments.Methods The effects of microgravity on osteoblasts were simulated by using a rotary cell culture system.MC3T3-E1 osteoblasts were transfected with an miR-194-3p inhibitor and changes in proliferation,differentiation,apoptosis,and mineralization were assessed using MTT assay,RT-PCR,Western blot,double fluorescence staining,and alizarin red staining.Results Elevated expression of miR-194-3p under simulated microgravity conditions led to the suppression of osteoblast proliferation,differentiation,and mineralization to a certain extent,while promoting osteoblast apoptosis.However,transfection with the miR-194-3p inhibitor significantly downregulated miR-194-3p expression and 623 partially reversed the reduced osteoblast proliferation,decreased expression of osteogenic differentiation markers such as ALP,OCN,and COL-I genes and proteins,decreased bone mineralization nodules,and increased osteoblast apoptosis induced by microgravity exposure.These findings indicated that miR-194-3p effectively ameliorates abnormal osteoblast function under microgravity conditions.Conclusions MiR-194-3p acts as a negative regulatory factor in the mechanical responses of osteoblasts under simulated microgravity.

关 键 词：模拟微重力环境 miR-194-3p 成骨细胞 细胞增殖 细胞分化 矿化 力学响应 

分 类 号：R318.01[医药卫生—生物医学工程]