纤维蛋白原诱导SH-SY5Y细胞凋亡和损伤的机制研究

A study on the mechanism of fibrinogen-induced apoptosis and mitochondrial damage in SH-SY5Y cells

目的探讨纤维蛋白原(Fibrinogen,Fg)对人神经母细胞瘤细胞(Human neuroblastoma cells,SH-SY5Y)的影响及其机制。方法采用不同水平的纤维蛋白原(0、2、4、8 mg/mL)处理SH-SY5Y细胞,并使用了多种实验方法来评估其对细胞的影响:CCK8试剂盒(Cell counting kit-8 viability assay)检测细胞活力;Hoechst33342/碘化丙啶(Propidine iodide,PI)染色试剂盒观察细胞凋亡情况;细胞色素c氧化酶IV(Cytochrome C oxidase IV,COX IV)细胞免疫荧光染色检测线粒体功能损伤;蛋白免疫印迹法检测线粒体凋亡通路相关指标[B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl2关联X蛋白(Bcl2 associated X protein,Bax)、细胞色素C(Cytochrome C,Cyt-C)、活化半胱胺酸蛋白酶蛋白-3(Activated caspase-3)]的表达水平。结果SH-SY5Y细胞暴露于纤维蛋白原会显著降低细胞活力,触发神经元凋亡,诱导线粒体功能障碍。此外,Western blot分析发现纤维蛋白原处理也改变了SH-SY5Y细胞中线粒体通路凋亡相关指标(Bcl-2,Bax,Cytochrome C,Activated caspase-3)的表达水平。结论纤维蛋白原引起了SH-SY5Y细胞的损伤和凋亡,其作用机制可能涉及线粒体功能障碍。

Objective To investigate the effect of fibrinogen(Fg)on human neuroblastoma cells(SH-SY5Y)and to elucidate the underlying mechanism.Methods SH-SY5Y cells were treated with graded concentrations of Fg(0,2,4,and 8 mg/mL)and the effects were assessed using a variety of methods:cell viability was assessed using the Cell Counting Kit-8 Viability Assay;the extent of apoptosis was assessed using the Hoechst33342/propidium iodide double staining;mitochondrial damage was assessed using COX IV cell immunofluorescence staining and immunoblotting of mitochondrial apoptotic pathway-related markers(Bcl-2,Bax,cytochrome c,activated caspase-3).Results Fg treatment resulted in a significant decrease in cell viability,exacerbated cell apoptosis,and induced mitochondrial dysfunction in SH-SY5Y cells in a concentration-dependent manner.Immunoblotting results indicated that Fg treatment may induce apoptosis by altering the expression levels of mitochondrial apoptotic pathway-related markers.Conclusion Fg caused damage and apoptosis in SH-SY5Y cells via a mechanism that may involve mitochondrial dysfunction.