摘要
目的:探讨氧化应激在高糖诱导成骨细胞凋亡中的作用。方法:体外培养的MC3T3-E1细胞,分为正常对照组、N-乙酰半胱氨酸(N-acetyl-L-cysteine,NAC)组、11.0 mmol/L高糖组、11.0 mmol/L高糖+NAC组、22.0 mmol/L高糖组、22.0 mmol/L高糖+NAC组。应用Annexin V-FITC/PI处理不同组别细胞,流式细胞术检测细胞凋亡;Honchest33258染色观察细胞核形态;采用MTT法检测细胞增殖;采用DCFH-DA荧光探针检测细胞活性氧类(reactive oxygen species,ROS)的生成。结果:111.0 mmol/L高糖组较正常对照组MC3T3-E1细胞凋亡率显著增加(P=0.004);22.0 mmol/L高糖组较正常对照组和11.0 mmol/L高糖组MC3T3-E1细胞凋亡率明显增加(P=0.000;P=0.000)。211.0 mmol/L高糖组较正常对照组MC3T3-E1细胞增殖轻微下降(P=0.305);与正常对照组和11.0 mmol/L高糖组相比,22.0 mmol/L高糖组MC3T3-E1细胞增殖显著降低(P=0.001;P=0.009)。311.0 mmol/L和22.0mmol/L高糖组MC3T3-E1细胞胞内ROS水平均明显增加(P=0.000;P=0.000),但无明显浓度依赖性,抗氧化剂NAC可明显降低ROS水平,改善高糖所致的凋亡增加和增殖下降。结论:高糖可通过增加成骨细胞胞内ROS生成引起MC3T3-E1成骨细胞凋亡增加、增殖下降。
Objective:To investigate the function of oxidative stress in apoptosis and proliferation of MC3T3-E1 cells under high glucose. Methods:MC3T3-E1 cells cultured in vitro were divided into normal control group,N-acetyl-L-cysteine(NAC) group,11.0mmol/L high-glucose group,11.0 mmol/L high-glucose+NAC group,22.0 mmol/L high-glucose group and 22.0 mmol/L high-glucose+NAC group. Reactive oxygen species(ROS) production was measured with DCFH-DA fluorescent probe. Cell proliferation was measured by MTT analysis. Cells in different groups were stained with Annexin V-FITC/PI,then the apoptosis rates were detected by flow cytometry. Morphology of cell nucleus was observed with Honchest33258. Results:1Cell apoptosis of 11.0 mmol/L high-glucose increased dramatically compared with that of normal control group(P=0.004). Cell apoptosis in 22.0 mmol/L high-glucose group increased significantly compared with that of normal control group and 11.0 mmol/L high-glucose group(P=0.000;P=0.000). 2Proliferation in 11.0 mmol/L high-glucose slightly decreased compared with that of normal control group(P=0.305). Compared with that of normal control and 11.0 mmol/L high-glucose group,proliferation in 22.0 mmol/L high-glucose group decreased significantly(P=0.001;P=0.009). 3ROS production in osteoblasts remarkablely increased under high glucose(P=0.000;P=0.000),but not in a dosedependent manner. NAC,as an antioxidant,could reduce ROS production obviously and ameliorate cell apoptosis and proliferation abnormality caused by high glucose. Conclusion :High glucose can increase ROS production in osteoblasts and subsequently lead to the increase of apoptosis and decrease of proliferation.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2015年第7期960-964,共5页
Journal of Chongqing Medical University
基金
国家临床重点专科建设项目资助项目(编号:2012)
重庆市卫生局科研基金资助项目(编号:12-02-041)
关键词
氧化应激
活性氧类
凋亡
增殖
oxidative stress
reactive oxygen species
apoptosis
proliferation