氟伐他汀调控Sirt3对过氧化氢诱导的骨髓间充质干细胞增殖凋亡的影响

Effect of fluvastatin on proliferation and apoptosis of bone marrow mesenchymal stem cells induced by hydrogen peroxide

目的探讨氟伐他汀对过氧化氢诱导的骨髓间充质干细胞(BMSCs)增殖凋亡的影响及作用机制。方法用400μmol/L的过氧化氢处理BMSCs作为模型组;BMSCs用400μmol/L的过氧化氢培养的同时分别加入0.01、0.1、1μmol/L氟伐他汀作为干预组;未添加任何物质正常培养的细胞作为正常组;将si-NC、si-Sirt3转染至BMSCs中再用400μmol/L的过氧化氢、0.1μmol/L氟伐他汀处理作为干预组2+si-NC组、干预组2+si-Sirt3组。蛋白质印迹(WesternBlotting)法检测Sirt3蛋白表达水平;四甲基偶氮唑盐比色法(MTT)检测细胞存活率;流式细胞术检测细胞凋亡。结果与正常组相比,模型组BMSCs细胞存活率降低[(41.57±4.32)%比(100.40±10.23)%],细胞凋亡率升高[(38.57±4.04)%比(8.28±0.92)%],Sirt3表达水平降低[(0.31±0.03)比(1.01±0.10)],均差异有统计学意义(P<0.05);与模型组相比,氟伐他汀干预组细胞存活率明显升高细胞凋亡率降低,Sirt3表达水平升高,均差异有统计学意义(P<0.05)。提示抑制Sirt3能逆转氟伐他汀对过氧化氢诱导的BMSCs细胞增殖抑制和凋亡促进作用。结论氟伐他汀能抑制过氧化氢诱导的BMSCs细胞凋亡,其机制可能与上调Sirt3有关。

Objective To investigate the effect and mechanism of fluvastatin on proliferation and apoptosis of bone marrow mesen⁃chymal stem cells(BMSCs)induced by hydrogen peroxide.Methods BMSCs were treated with 400μmol/L hydrogen peroxide as a model group;BMSCs were cultured with 400μmol/L hydrogen peroxide while adding 0.01,0.1,1μmol/L fluvastatin as intervention group;normal cultured cells without any additives served as normal group;BMSCs transfecte with si-NC and si-Sirt3 and treated with 400μmol/L hydrogen peroxide and 0.1μmol/L fluvastatin were designated as intervention group 2+si-NC group,intervention group 2+si-Sirt3 group.The expression of Sirt3 protein was detected by Western Blot;cell viability was detected by MTT assay;cell apoptosis was detected by flow cytometry.Results Compared with normal group,the cell survival rate of BMSCs in model group was significant⁃ly decreased[(41.57±4.32)%vs.(100.40±10.23)%],the apoptosis rate was significantly increased[(38.57±4.04)%vs.(8.28±0.92)%],and the expression of Sirt3 was significantly decreased[(0.31±0.03)vs.(1.01±0.10)](P<0.05);Compared with model group,the cell survival rate of the intervention group was significantly increased,and the apoptosis rate was significantly decreased.The expression of Sirt3 was significantly increased(P<0.05).Inhibition of Sirt3 reversed the effect of fluvastatin on proliferation inhibition and apoptosis of BMSCs induced by hydrogen peroxide.Conclusion Fluvastatin can inhibit the apoptosis of BMSCs induced by hydrogen peroxide,and its mechanism may be related to up-regulation of Sirt3.