波动性高糖对内皮细胞自噬基因表达的影响  

Effects of fluctuant hyperglycemia on autophagy of endothelial cells

作　　者：洪新宇[1] 杨正丽 马运芹 张明亮[2] 肖萍[1] HONG Xin-yu;YANG Zheng-li;MA Yun-qin;ZHANG Ming-liang;XIAO Ping(Department of In Vivo Toxicology Test and Evaluation,Shanghai Municipal Center for Disease Control and Prevention,Shanghai 200336,China;Department of Endocrinology and Metabolism,Shanghai Sixth People's Hospital,Shanghai 200233,China)

机构地区：[1]上海市疾病预防控制中心动物毒理检测与评价室,上海200336 [2]上海市第六人民医院内分泌代谢科,上海200233

出　　处：《上海预防医学》2020年第3期248-252,共5页Shanghai Journal of Preventive Medicine

基　　金：上海市卫生健康委员会面上项目(201540287)。

摘　　要：【目的】探讨波动性高糖对内皮细胞自噬的影响及其可能的机制。【方法】体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)用于实验。实验分正常对照组(5.5 mmol/L葡萄糖)和葡萄糖交替处理组(5.5/28 mmol/L葡萄糖)。其中葡萄糖交替处理模拟人血糖波动状态,方案是前12 h内每隔4 h交替培养,后12 h内葡萄糖维持5.5 mmol/L水平,按此循环,共培养48 h。用CCK-8检测细胞活性,酶标仪检测氧化应激相关指标,实时荧光定量PCR检测自噬相关基因的表达情况。【结果】与正常对照组相比,葡萄糖交替处理组降低HUVEC细胞活性(P<0.01);降低细胞内抗氧化酶活性,增加丙二醛和8-羟基脱氧鸟苷的含量(P<0.01);增加自噬相关基因Atg4和Atg5的表达(P<0.001)。【结论】波动性高糖可能是先通过诱导HUVEC产生氧化应激,进而使HUVEC产生自噬,最终降低细胞活性。[Objective]To investigate the effect of fluctuant hyperglycemia on autophagy of endothelial cells and its possible mechanism.[Methods]Human umbilical vein endothelial cells(HUVEC)were cultured in vitro for experiments with control group(5.5 mmol/L glucose)and intermittent high glucose group(5.5/28 mmol/L glucose).Intermittent high glucose simulating human blood glucose fluctuations was alternately cultured every 4 hours in the first 12 h and 5.5 mmol/L for another 12 h.According to this cycle,the cells were cultured for 48 h in total.Cell viability was measured by CCK-8,indicators related oxidative stress were detected by microplate reader,and detection of gene related autophagy by real-time fluorescent quantitative PCR.[Results]Compared with the control group,intermittent high glucose group reduced cell viability,decreased intracellular antioxidant enzyme activity,increased the expression of malondialdehyde(MDA)and 8-Hydroxydeoxyguanosine(8-OHdG)(P<0.01),and increased the expression of Atg4 and Atg5 related autophagy(P<0.001).[Conclusion]Fluctuant hyperglycemia may induce oxidative stress of HUVEC,thereby causing autophagy of HUVEC,ultimately to reduce cell viability.

关 键 词：糖尿病 波动性高糖 内皮细胞 氧化应激 自噬 

分 类 号：R114[医药卫生—卫生毒理学] R587.1[医药卫生—公共卫生与预防医学]