晚期氧化蛋白产物诱导HaCaT细胞凋亡的实验研究  

Advanced oxidation protein products induce apoptosis of HaCaT cells: an experimental study

作　　者：孙百慧[1] 李勤[1] 丁若汀 余文林[2] 吴燕虹[2] 

机构地区：[1]广州南方医科大学研究生学院,510515 [2]广州军区广州总医院激光整形中心,广州510010

出　　处：《解放军医学杂志》2016年第5期373-377,共5页Medical Journal of Chinese People's Liberation Army

基　　金：广东省科技计划项目(2013B040401013)~~

摘　　要：目的探讨晚期氧化蛋白产物(AOPPs)对HaCaT细胞凋亡的影响及其可能的作用机制。方法人血清白蛋白(HSA)与次氯酸钠溶液反应制成AOPPs。MTT法检测经不同浓度(0、50、100、200、400μg/ml)AOPPs处理24h后HaCaT细胞的活力,以选择AOPPs的工作浓度。根据时间效应或浓度效应对HaCaT细胞进行分组处理,流式细胞技术检测细胞凋亡率,二氯荧光素双醋酸盐(DCFH-DA)法检测细胞内活性氧(ROS)水平,Western blotting技术检测细胞内NOX4、Bax及Bcl-2蛋白的表达。结果 400μg/ml AOPPs作用后HaCaT细胞活力较其他浓度明显降低(P<0.05)。随AOPPs作用时间的延长和浓度的增加,HaCaT细胞凋亡率明显增加(P<0.05),细胞内ROS产生增多(P<0.05),NOX4和Bax蛋白表达量增加(P<0.05),Bcl-2蛋白表达量降低(P<0.05)。结论 AOPPs可能通过NADPH氧化酶途径诱导HaCaT细胞凋亡。Objective To investigate the effect and mechanism of advanced oxidation protein products（AOPPs） on the apoptosis of HaCaT cells. Methods AOPPs were prepared by incubation of human serum albumin（HSA） with sodium hypochlorite solution. MTT assay was used to determine the effects of different concentrations（0, 50, 100, 200 and 400μg/ml） of AOPPs on HaCaT cell activity by incubating the cells for 24 h for selecting optimum working concentration of AOPPs. HaCaT cells were treated based on time effect or concentration effect. The incidence of cell apoptosis was measured by flow cytometry. Intracellular reactive oxygen species（ROS） production was determined with dichlorofluorescein diacetate（DCFH-DA）. And the protein expressions of NOX4, Bax and Bcl-2 were examined by Western blotting. Results Viability of HaCaT cells treated with 400μg/ml AOPPs was significantly lowered（P〈0.05）. AOPPs, in a time-and concentration-dependent manner, induced the apoptosis of HaCaT cells（P〈0.05）, increased the expression of NOX4 and Bax（P〈0.05） and down-regulated the expression of Bcl-2（P〈0.05）; prolonged exposure to AOPPs and increase of AOPPs concentration both led to an increase of intracellular ROS production（P〈0.05）. Conclusion AOPPs may induce apoptosis of HaCaT cells through NADPH oxidase pathway.

关 键 词：晚期氧化蛋白产物 HACAT细胞 活性氧 细胞凋亡 

分 类 号：R644.02[医药卫生—外科学]