摘要
目的:探讨飞燕草素葡萄糖苷(DPg)对高糖(HG)诱导的心肌细胞H9C2损伤的影响及可能机制。方法:体外培养H9C2细胞,不同剂量(1,10,100μmol·L^(-1))的DPg作用于33.3 mmol·L^(-1)葡萄糖诱导的H9C2细胞24 h,CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,Western Blot检测细胞中兔抗鼠B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关蛋白(Bax)的蛋白表达,试剂盒检测细胞中丙二醛(MDA)和超氧化物歧化酶(SOD)水平,RT-qPCR检测SRY盒转录因子2重叠转录本(SOX2OT)表达。转染SOX2OT过表达载体、小干扰RNA至H9C2细胞,经33.3 mmol·L^(-1)葡萄糖诱导24 h后,检测细胞增殖、凋亡、Bax和Bcl-2蛋白表达及MDA和SOD水平。结果:DPg可提高高糖诱导的H9C2细胞吸光度(A)值(P<0.05),降低细胞凋亡率、Bax蛋白表达及MDA含量(P<0.05),并促进Bcl-2蛋白表达和SOD活性(P<0.05),且呈剂量依赖性。DPg可剂量依赖性促进高糖诱导的H9C2细胞中SOX2OT的表达,过表达SOX2OT可提高高糖诱导的H9C2细胞A值(P<0.05),降低细胞凋亡率、Bax蛋白表达及MDA含量(P<0.05),并促进Bcl-2蛋白表达和SOD活性(P<0.05)。与未敲减SOX2OT的H9C2细胞比较,敲减SOX2OT的H9C2细胞经100μmol·L^(-1)DPg和高糖处理后A值、Bcl-2蛋白表达和SOD活性降低(P<0.05),细胞凋亡率、Bax蛋白表达及MDA含量升高(P<0.05),并抑制Bcl-2蛋白表达和SOD活性(P<0.05)。结论:DPg可能通过上调SOX2OT表达抑制HG诱导的H9C2细胞凋亡和氧化应激。
Objective:To investigate the effects of delphinidin-glucoside(DPg)on cardiomyocytes H9C2 injury induced by high glucose(HG)and its possible mechanism.Methods:H9C2 cells were cultured in vitro.Different doses(1,10,100μmol·L^(-1))of DPg were applied to H9C2 cells induced by 33.3 mmol·L^(-1) glucose for 24 h.Cell proliferation was detected by CCK-8 method,cell apoptosis was detected by flow cytometry,the expressions of Blot and Bax protein were detected by Western blot,the levels of MDA and SOD in cells were detected by kits,and the expression of SOX2OT was detected by RT-qPCR.After the SOX2OT overexpression vector or small interfering RNA was transfected into H9C2 cells,they were induced with 3.3 mmol·L^(-1) glucose for 24 hours,and then the cell proliferation,apoptosis,expressions of Bax and Bcl-2 protein,and levels of MDA and SOD were detected by the same above methods.Results:DPg could increase the absorbance(A)value of H9C2 cells induced by high glucose(P<0.05),reduce the apoptosis rate,expression of Bax protein and content of MDA(P<0.05),and promote the expression of Bcl-2 protein and SOD activity(P<0.05),and all the effects were dose-dependent.DPg could promote the expression of SOX2OT in H9C2 cells induced by high glucose in a dose-dependent manner.Overexpression of SOX2OT could increase the A value of H9C2 cells induced by high glucose(P<0.05),reduce the rate of apoptosis,expression of Bax protein and content of MDA(P<0.05),and promote the expression of Bcl-2 protein and SOD activity(P<0.05).Compared with HPC2 cells without SOX2OT knockdown,and A value,the expression of Bcl-2 protein and SOD activity of H9C2 cells with SOX2OT knockdown treated with 100μmol·L^(-1) DPg and HG were decreased(P<0.05),and the apoptosis rate,expression of Bax protein and content of MDA were in ceased(P<0.05),and inhibited the expression of Bcl-2 protein and SOD activity(P<0.05).Conclusion:DPg may inhibit apoptosis and oxidative stress of HG-induced H9C2 cells by up-regulating SOX2OT.
作者
庄媛
李延辉
卢云凤
王位坐
安宁
郝艳丽
Zhuang Yuan;Li Yanhui;Lu Yunfeng;Wang Weizuo;An Ning;Hao Yanli(Department of Cardiology,Jinqiu Hospital of Liaoning Province,Shenyang 110015,China;Department of Neurosurgery,the Fourth Affiliated Hospital of China Medical University)
出处
《中国药师》
CAS
2021年第7期228-234,共7页
China Pharmacist
基金
辽宁省自然科学基金指导计划立项项目(编号:201602413)。
