维生素D3减轻高糖暴露诱导氧化应激促进人脐带间充质干细胞的成骨分化  

Vitamin D3 attenuates high-glucose exposure-induced oxidative stress to promote osteogenic differentiation of human umbilical cord mesenchymal stem cells

作　　者：谢婷[1] 刘婷婷 曾雪慧 李亚敏 周庞虎[2] 易念华[1] Xie Ting;Liu Tingting;Zeng Xuehui;Li Yamin;Zhou Panghu;Yi Nianhua(Department of Women’s Health Care,Maternal and Child Health Hospital of Hubei Province,Wuhan 430070,Hubei Province,China;Department of Orthopedics,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei Province,China)

机构地区：[1]湖北省妇幼保健院妇女保健科,湖北省武汉市430070 [2]武汉大学人民医院骨科,湖北省武汉市430060

出　　处：《中国组织工程研究》2024年第19期2981-2987,共7页Chinese Journal of Tissue Engineering Research

基　　金：湖北省重点研发计划项目(2021BCA147),项目负责人:周庞虎。

摘　　要：背景:糖尿病骨质疏松症逐渐受到公众的关注,然而鲜有研究报道高糖环境对人脐带间充质干细胞成骨分化的影响和相应的治疗策略。目的:探讨维生素D3恢复高糖环境中人脐带间充质干细胞成骨分化的潜力。方法:通过CCK-8法检测人脐带间充质干细胞活力来筛选适宜的维生素D3干预浓度。在高葡萄糖条件下,通过RT-qPCR、蛋白质印迹、免疫荧光、JC-1线粒体膜电位、茜素红染色和β-半乳糖苷酶染色等实验评估维生素D3干预后人脐带间充质干细胞的成骨分化潜能、细胞内活性氧积累、线粒体膜电位改变和细胞衰老情况,并探讨了潜在的机制。结果与结论:①维生素D3在0.1μmol/L至1 mmol/L范围内均能显著促进人脐带间充质干细胞的增殖;②高糖环境下调了人脐带间充质干细胞中成骨相关基因α1-Ⅰ型胶原蛋白、碱性磷酸酶、Runt相关转录因子2、骨钙蛋白的mRNA和蛋白表达水平,诱发了氧化应激和细胞衰老;③维生素D3在10μmol/L的干预浓度下显著恢复了高糖条件下人脐带间充质干细胞的成骨表型,并通过激活Nrf2/HO-1信号通路减轻了细胞内的氧化应激和细胞衰老;④结果表明,高糖环境中人脐带间充质干细胞的成骨分化能力降低,维生素D3可以部分提高其成骨分化能力并减轻细胞损伤。BACKGROUND:Diabetic osteoporosis is gaining public attention.However,few studies have reported the effect of a high-glucose environment on the osteogenic differentiation of human umbilical cord mesenchymal stem cells and the corresponding therapeutic strategies.OBJECTIVE:To investigate whether vitamin D3 can restore the osteogenic differentiation potential of human umbilical cord mesenchymal stem cells in a high-glucose environment.METHODS:The viability of human umbilical cord mesenchymal stem cells was detected by CCK-8 assay to screen the appropriate vitamin D3 intervention concentration.Under the high-glucose environment,RT-qPCR,western blot assay,immunofluorescence,JC-1 mitochondrial membrane potential,alizarin red staining,andβ-galactosidase staining were used to evaluate the osteogenic differentiation potential,intracellular reactive oxygen species accumulation,mitochondrial membrane potential alteration,and cell senescence of human umbilical cord mesenchymal stem cells after vitamin D3 intervention.The underlying mechanism was also discussed.RESULTS AND CONCLUSION:(1)Vitamin D3 significantly promoted the proliferation of human umbilical cord mesenchymal stem cells in the range of 0.1μmol/L to 1 mmol/L.(2)High-glucose environment down-regulated the mRNA and protein level expressions of osteogenic-related genesα1-I collagen,alkaline phosphatase,Runt-associated transcription factor 2,and osteocalcin in human umbilical cord mesenchymal stem cells,which induced oxidative stress and cellular senescence.(3)Vitamin D3 at an intervention concentration of 10μmol/L significantly restored the osteogenic phenotype of human umbilical cord mesenchymal stem cells under high-glucose conditions and attenuated intracellular oxidative stress and cellular senescence by activating the Nrf2/HO-1 signaling pathway.(4)These findings suggested that the osteogenic differentiation ability of human umbilical cord mesenchymal stem cells was reduced in the high-glucose environment,and vitamin D3 could partially improve their osteoge

关 键 词：糖尿病骨质疏松症 人脐带间充质干细胞 维生素D3 高血糖 线粒体功能障碍 活性氧 细胞衰老 成骨分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R58