目的 研究鸢尾素(irisin)对炎症状态下骨髓间充质干细胞(BMSCs)成骨分化的保护作用。方法 分离、培养小鼠BMSCs并将其分为3组:对照组(Vehicle),牙龈卟啉单胞菌脂多糖诱导炎症组(LPS),Irisin干预组(LPS+irisin;Irisin)。CCK-8试剂盒检测细胞活力;成骨诱导培养后检测碱性磷酸酶(ALP)活性,qPCR检测成骨相关基因Runt相关转录因子2(Runx2)、锌指结构转录因子(Osx)、ALP和骨钙素(OCN)的转录表达,Western blot检测成骨相关蛋白ALP和OCN的表达,茜素红染色半定量分析检测矿化水平。结果 与Vehicle组相比,LPS显著促进BMSCs增殖(P＜0.05);LPS下调Runx2、Osx、ALP和OCN的转录水平,降低ALP和OCN蛋白表达(P＜0.05),而irisin干预显著上调Runx2、Osx和ALP转录表达,提高ALP和OCN蛋白表达,并提高矿化水平(P＜0.05)。结论 Irisin可促进炎症微环境中小鼠BMSCs成骨分化。
Abstract
Objective To investigate the effects of irisin on osteoblast differentiation of bone-marrow mesenchymal stem cells (BMSCs) in the inflammatory microenvironment.Methods BMSCs were isolated, cultured and divided into 3 groups: Vehicle, LPS and Irisin (LPS plus irisin intervention). CCK-8 was used to access cell proliferation. After cultured in osteogenic differentiation medium, ALP activity was evaluated, mRNA levels of Runx2, Osx, ALP and OCN and protein levels of ALP and OCN were also investigated. Moreover, alizarin red was used to determine mineralization level.Results Compared with the Vehicle group, LPS significantly promoted BMSCs proliferation (P＜0.05); LPS downregulated Runx2, Osx, ALP and OCN mRNA expression and reduced ALP and OCN protein levels, while irisin intervention could obviously upregulate Runx2, Osx and ALP mRNA expression, enhanced ALP and OCN protein levels, and raised mineralization level (P＜0.05). Conclusions Irisin can promote osteoblast differentiation of BMSCs in the inflammatory microenvironment.
关键词
鸢尾素 /
炎症微环境 /
骨髓间充质干细胞 /
成骨分化
Key words
irisin /
inflammatory microenvironment /
bone-marrow mesenchymal stem cells /
osteoblast differentiation
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基金
博士后科学基金资助项目(2019T120977)
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