辅酶Q10对高糖引起的人冠状动脉内皮细胞凋亡的抑制作用及其机制

doi:10.13481/j.1671-587x.20190510

摘要/Abstract

摘要： 目的：探讨辅酶Q10（Co-Q10）对高糖诱导的人冠状动脉内皮细胞（HCAECs）凋亡的抑制作用，并阐明其可能的作用机制。方法： HCAECs分为对照组，高糖组，高糖联合5、10和20 μmol·L^-1 Co-Q10处理组。对照组HCAECs采用常规培养方法培养24 h；高糖组采用30 mmol·L^-1葡萄糖处理细胞24 h；高糖联合5、10和20 μmol·L^-1 Co-Q10处理组分别采用5、10和20 μmol·L^-1 Co-Q10联合30 mmol·L^-1葡萄糖处理细胞24 h。采用CCK-8法检测各组细胞活性，Hoechst-PI双染色法检测高糖组和高糖联合10 μmol·L^-1 Co-Q10处理组细胞凋亡率，Mito-tracker染色检测高糖组和高糖联合10 μmol·L^-1 Co-Q10处理组细胞线粒体膜电位，MitoSOX染色检测高糖组和高糖联合10 μmol·L^-1 Co-Q10处理组细胞线粒体活性氧（mtROS）水平，Western blotting法检测高糖组和高糖联合10 μmol·L^-1 Co-Q10处理组细胞中B细胞淋巴瘤/白血病2蛋白（Bcl-2）、Bcl-2相关X蛋白（Bax）、Bcl-2相关死亡启动子（Bad）和X连锁凋亡抑制蛋白（x-IAP）表达水平。结果：与对照组比较，高糖组细胞活性明显降低（P<0.01）；与高糖组比较，高糖联合5、10和20 μmol·L^-1 Co-Q10处理组细胞活性均明显升高（P<0.05或P<0.01），高糖联合10 μmol·L^-1 Co-Q10处理组升高最为明显（P<0.01）。与高糖组比较，高糖联合10 μmol·L^-1 Co-Q10处理组细胞凋亡率、线粒体膜电位和mtROS水平均明显降低（P<0.01）。Western blotting法检测，与高糖组比较，高糖联合10 μmol·L^-1 Co-Q10处理组HCAECs中Bax和Bad表达水平明显降低（P<0.01），Bcl-2和x-IAP表达水平均明显升高（P<0.01）。结论： Co-Q10可能通过抑制线粒体凋亡相关通路减少高糖引起的HCAECs凋亡，对细胞起保护作用。

关键词: 辅酶Q10, 高糖, 人冠状动脉内皮细胞, 细胞凋亡, 线粒体应激

Abstract: Objective:To investigate the inhibitory effect of coenzyme Q10 (Co-Q10) on the apoptosis of human coronary endothelial cells (HCAECs) induced by highglucose, and to elucidate its possible mechanism. Methods:The HCAECs were divided into control group, high glucose group and high glucose combined with 5, 10,and20 μmol·L^-1 Co-Q10 treatment groups;the HCAECs in control group were cultured for 24 h using a routine culture method. The cells in high glucose group were treated with 30 mmol·L^-1 glucose for 24 h;the cells in high glucose combined with 5, 10,and 20 μmol·L^-1 Co-Q10 treatment groups were treated with 5, 10,and 20 μmol·L^-1 Co-Q10 combined with 30 mmol·L^-1 glucose for 24 h, respectively. The cell viabilities of HCAECs in various groups were measured by CCK-8 assay. The apoptotic rates of HCAECs in high glucose group and high glucose combined with 10 μmol·L^-1 Co-Q10 treatment group were detected by Hoechst-PI double staining. The cell mitochondrial membrane potentials of HCAECs in high glucose group and high glucose combined with 10 μmol·L^-1 Co-Q10 treatment group were determined by Mito-tracker staining.The mitochondrial reative oxygen species (mtROS) levels in the HCAECs in high glucose group and high glucose combined with 10 μmol·L^-1 Co-Q10 treatment group were measured by MitoSox staining. The protein expression levels of B cell lymphoma/leukemia 2 protein (Bcl-2), Bcl-2 assaciated X protein (Bax), Bcl-2 assaciated death promoter (Bad) and X-linked inhibitor of apoptosis protein (x-IAP) in the HCAECs in high glucose group and high glucose combined with 10 μmol·L^-1 Co-Q10 treatment group were detected by Western blotting method. Results:Compared with control group, the cell viability of HCAECs in high glucose group was significantly reduced (P<0.01); compared with high glucose group, the cell viabilities of HCAECs in high glucose combined with 5, 10,and 20 μmol·L^-1 Co-Q10 treatment groups were significantly increased(P<0.05 or P<0.01), especially in high glucose combined with 10 μmol·L^-1 Co-Q10 treatment group (P<0.01). Compared with high glucose group, the apoptotic rate, the mitochondrial membrane potential and the mtROS level of HCAECs in high glucose combined with 10 μmol·L^-1 Co-Q10 treatment group were significantly decreased (P<0.01).The Western blotting results showed that compared with high glucose group, the expression levels of Bax and Bad proteins in the HCAECs in high glucose combined with 10 μmol·L^-1 Co-Q10 treatment group were decreased significantly(P<0.01), and the expression levels of Bcl-2 and x-IAP proteins were increased significantly (P<0.01). Conclusion:Co-Q10 may reduce the apoptosis of HCAECs induced by high glucose through inhibiting the mitochondrial apoptosis-related pathway to ptotect the cells.

Key words: coenzyme Q10, high glucose, human coronary endothelial cells, apoptosis, mitochondrial stress

中图分类号:

R543.3

郭青榜, 冯文化, 张钊. 辅酶Q10对高糖引起的人冠状动脉内皮细胞凋亡的抑制作用及其机制[J]. 吉林大学学报(医学版), 2019, 45(05): 1031-1035.

GUO Qingbang, FENG Wenhua, ZHANG Zhao. Inhibitory effect of coenzyme Q10 on apoptosis of human coronary endothelial cells induced by high glucose and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2019, 45(05): 1031-1035.

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