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
薛进冬 1程路畅 1王敏 1谭彩美 1邓琪琪 1朱海美 1郭勇 2韩标2
作者信息
- 1. 桂林医学院 智能医学与生物技术学院,桂林 541199
- 2. 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
- 折叠
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.
Abstract
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.
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Key words
simulated microgravity/miR-194-3p/osteoblast/cell proliferation/cell differentiation/mineralization/mechanical response引用本文复制引用
基金项目
广西自然科学基金(2020GXNSFBA238006)
广西自然科学基金(桂科AD22080037)
国家自然科学基金(12202113)
国家自然科学基金(32071309)
广西壮族自治区硕士研究生创新项目(YCSW2022368)
出版年
2024
NETL
NSTL
万方数据