首页|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

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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 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.

simulated microgravitymiR-194-3posteoblastcell proliferationcell differentiationmineralizationmechanical response

薛进冬、程路畅、王敏、谭彩美、邓琪琪、朱海美、郭勇、韩标

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桂林医学院 智能医学与生物技术学院,桂林 541199

广西高校医药生物技术与转化医学重点实验室,桂林 541199

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

广西自然科学基金广西自然科学基金国家自然科学基金国家自然科学基金广西壮族自治区硕士研究生创新项目

2020GXNSFBA238006桂科AD220800371220211332071309YCSW2022368

2024

10.16156/j.1004-7220.2024.04.008

医用生物力学
上海第二医科大学

医用生物力学

CSTPCD北大核心
影响因子：0.858
ISSN：1004-7220
年,卷(期)：2024.39(4)