骨髓间充质干细胞成脂分化介导酒精性股骨头缺血性坏死发生机制的实验研究

Experimental study on mechanism of alcoholic femoral head necrosis mediated by adipogenic differentiation of bone marrow mesenchymal stem cells

目的探讨骨髓间充质干细胞(Bone mesenchymal stem cells,BMSCs)成脂分化对大鼠酒精性股骨头缺血性坏死的影响及其发生机制。方法构建酒精诱导的大鼠股骨头缺血性坏死模型,进行大鼠股骨头标本HE染色及免疫组织化学染色,同时检测LncRNA-HOTAIR/miR-122/PPARγ信号表达变化。采用不同剂量(0、10、50、100 mM)的乙醇(80%浓度)处理大鼠BMSCs 72 h,然后检测BMSCs的成脂和成骨分化能力,以及LncRNA-HOTAIR/miR-122/PPARγ信号表达变化。结果动物实验结果显示,与对照组相比,乙醇组中miR-122的表达降低,而LncRNA-HOTAIR的表达增加,PPARγ的蛋白表达显著增加(P<0.05)。细胞实验结果显示,乙醇以剂量依赖性方式增加了FABP4、FAS、LPL的mRNA表达(P<0.05);碱性磷酸酶染色结果显示,乙醇以剂量依赖性的方式抑制细胞外矿化的形成(P<0.05);乙醇可剂量依赖性方式抑制miR-122的mRNA表达,并促进LncRNA-HOTAIR和PPARγ的表达(P<0.05)。结论乙醇可能通过促进LncRNA-HOTAIR的表达抑制miR-122的表达,进而导致PPARγ的高表达,最终在乙醇诱导的股骨头缺血性坏死过程中介导BMSCs的脂肪分化。

Objective To investigate the effect of adipogenic differentiation of bone mesenchymal stem cells(BMSCs)on alcoholic avascular necrosis of femoral head in rats and its mechanism.Methods The model of alcoholic avascular necrosis of femoral head in rats was established.HE staining and immunohistochemical staining were performed on the specimen of rat femoral head and the LncRNA-HOTAIR/miR-122/PPARγsignal expression changes were detected.Rat BMSCs were treated with 80%ethanol of different doses(0,10,50,100 mM)for 72 hours,and the adipogenic and osteogenic differentiation abilities of BMSCs were detected respectively,as well as LncRNA-HOTAIR/miR-122/PPARγsignal expression changes.ResultsCompared with the control group,the expression of miR122 in the ethanol group decreased,while the expression of LncRNA-HOTAIR increased(P0.05).The protein expression of PPARγwas significantly increased(P0.05).The results of cell experiment showed that ethanol increased the mRNA expressions of FABP4,FAS and LPL in a dose-dependent manner(P0.05).ALP staining showed that ethanol inhibited the formation of extracellular mineralization in a dose-dependent manner(P0.05).It was also found that ethanol could inhibit the mRNA expression of miRNA122 in a dose-dependent manner,and promote LncRNA-HO-TAIR and PPARγ.ConclusionEthanol may inhibit the expression of miR122 by promoting the expression of LncRNA-HO-TAIR,thus leading to PPARγand finally mediates the adipose differentiation of BMSCs in the process of ethanol induced femoral head necrosis.