1,25(OH)_(2)D_(3)通过Bcl-2/Bax/cleaved caspase-3凋亡机制抑制小鼠气道平滑肌细胞的增殖  

1,25-Dihydroxy vitamin D3 attenuates the proliferation of mouse airway smooth muscle cells via Bcl-2/Bax/cleaved caspase-3 signaling pathway

作　　者：李飞 叶春雨 谭博 李胜文[2] Li Fei;Ye Chunyu;Tan Bo;Li Shengwen(Department of Pulmonary and Critical Care Medicine,Shanxi Provincial People′s Hospital,Taiyuan 030012,China;不详)

机构地区：[1]山西省人民医院呼吸与危重症医学科,太原030012 [2]山西省人民医院肿瘤科,太原030012 [3]山西医科大学第五临床医学院

出　　处：《山西医药杂志》2023年第3期169-172,共4页Shanxi Medical Journal

基　　金：山西省应用基础研究项目计划(201701-D221264)。

摘　　要：目的探讨1,25(OH)_(2)D_(3)对过氧化氢(H_(2)O_(2))刺激的小鼠气道平滑肌细胞(ASMCs)的抗增殖及促凋亡作用,并分析其在Bcl-2/Bax/cleaved caspase-3凋亡机制发挥的调控作用。方法体外培养小鼠气道平滑肌细胞,再用不同浓度(0、25、50、100μmol/L)的H_(2)O_(2)刺激细胞增殖,作用不同时间(12、24、48、72 h),探索H_(2)O_(2)最佳作用浓度和时间。接着将ASMCs分为对照组、H_(2)O_(2)组、1,25(OH)_(2)D_(3)、H_(2)O_(2)+1,25(OH)_(2)D_(3)4组进行培养,用四甲基偶氮唑盐(MTT)法检测各组ASMCs增殖活性的变化,并用流式细胞仪检测其凋亡情况。并用蛋白印迹法检测各组Bax、Bcl-2及cleaved caspase-3蛋白的表达情况。结果H_(2)O_(2)刺激小鼠气道平滑肌细胞增殖的最佳作用浓度和时间分别为25μmol/L和24 h。1,25(OH)_(2)D_(3)干预能明显降低ASMCs增殖能力,并增加其凋亡率。H_(2)O_(2)+1,25(OH)_(2)D_(3)组细胞凋亡率为21.9%(对照组细胞凋亡率为7.1%,H_(2)O_(2)组细胞凋亡率为4.8%),差异具有统计学意义(P<0.01)。1,25(OH)_(2)D_(3)干预后,蛋白印迹法结果显示ASMCs的Bax和cleaved Caspase-3蛋白表达水平升高,Bcl-2表达水平下降。结论1,25(OH)_(2)D_(3)能抑制H_(2)O_(2)刺激ASMCs的增殖,并促进其凋亡,其机制与Bax/Bcl-2/ccleaved caspase-3凋亡通路有关。Objective The aim of this study was to investigate the anti-proliferative and pro-apoptotic effects of 1,25(OH)_(2)D_(3) on H_(2)O_(2)-stimulated mouse airway smooth muscle cells arterial smooth muscle smooth(ASMCs),and to study its role in the Bcl-2/Bax/cleaved caspase-3 apoptotic pathway.Methods The proliferation of mouse ASMCs was determined to optimize the concentration and time of H_(2)O_(2)′s action.Subsequently,ASMCs were divided into four groups,which were treated for 24 h with ethanol vehicle,or 1,25(OH)_(2)D_(3) in the absence or presence of H_(2)O_(2).Then MTT assay was performed to monitor the proliferation of ASMCs in each group,and flow cytometry was conducted for apoptosis detection.The expressions of Bax,Bcl-2 and cleaved caspase-3 proteins were detected by Western-blot analysis.Results 1,25(OH)_(2)D_(3) intervention significantly reduced the proliferation and increased the apoptosis of ASMCs.Western blot analysis revealed that the expression levels of Bax and cleaved caspase-3 protein increased,and Bcl-2 protein decreased in ASMCs by 1,25(OH)_(2)D_(3) intervention.Conclusion 1,25(OH)_(2)D_(3) inhibits the proliferation of H_(2)O_(2)-stimulated ASMCs and promotes apoptosis via the Bax/Bcl-2/cleaved caspase-3 apoptotic pathway.

关 键 词：骨化三醇 细胞凋亡 细胞增殖 肌细胞 平滑肌 小鼠 Bcl-2 Bax CASPASE-3 

分 类 号：R562.25[医药卫生—呼吸系统]