甘草苷调节丙酮酸激酶M2改善过氧化氢诱导的人脐静脉内皮细胞功能障碍

Glycyrrhizin Regulates Pyruvate Kinase M2 to Improve Hydrogen Peroxide Induced Dysfunction of Human Umbilical Vein Endothelial Cells

目的:探讨甘草苷(LIQ)调节丙酮酸激酶M2(PKM2)对过氧化氢(H_(2)O_(2))诱导的人脐静脉内皮细胞(HUVECs)功能障碍的影响。方法:使用H_(2)O_(2)诱导HUVECs损伤模型,将细胞分为对照组(常规培养的HUVECs)、H_(2)O_(2)组(400μmol/L H_(2)O_(2)诱导的HUVECs模型)、H_(2)O_(2)+LIQ组(400μmol/L H_(2)O_(2)+1μmol/L LIQ)、H_(2)O_(2)+LIQ+紫草素(PKM2抑制剂)组(400μmol/L H_(2)O_(2)+1μmol/L LIQ+1μmol/L紫草素)。使用qRT-PCR检测PKM2 mRNA水平,荧光探针检测活性氧(ROS)水平,Western Blotting检测Nrf2、HO-1、PKM2、p-PKM2、Bcl-2、Bax及TNF-α、IL-1β蛋白表达变化。结果:与H_(2)O_(2)组相比,LIQ干预显著上调PKM2 mRNA水平、PKM2及p-PKM2的蛋白水平,抑制了ROS水平及TNF-α、IL-1β蛋白表达水平,Bcl-2/Bax比例显著增加。与H_(2)O_(2)+LIQ组相比,使用紫草素显著逆转了上述指标变化趋势(P<0.01)。结论:LIQ可上调PKM2表达,改善H_(2)O_(2)诱导的HUVECs细胞功能障碍。

Objective:To investigate the effects of glycyrrhizin(LIQ)regulating pyruvate kinase M2(PKM2)on hydrogen peroxide(H_(2)O_(2))-induced dysfunction of human umbilical vein endothelial cells(HUVECs).Method:HUVECs were divided into control group,H_(2)O_(2)group(400μmol/L H_(2)O_(2)),H_(2)O_(2)+LIQ group(400μmol/L H_(2)O_(2)+1μmol/L LIQ),and H_(2)O_(2)+LIQ+shikonin(PKM2 inhibitor)group(400μmol/L H_(2)O_(2)+1μmol/L LIQ+1μmol/L shikonin).qRT-PCR was used to detect the expression of PKM2 mRN.Silver light probe was used to detect the expression of ROS levels.Western Blot was used to detect the expression of Nrf2,HO-1,PKM2,p-PKM2,Bcl-2,Bax and TNF-α,IL-1β.Results:LIQ pretreatment significantly upregulated PKM2 mRNA levels,PKM2 and p-PKM2 protein levels.Compared with the H_(2)O_(2)group,LIQ pretreatment significantly inhibited ROS levels and the expression levels of TNF-αand IL-1βproteins,and significantly increased the Bcl-2/Bax ratio.Compared with the H_(2)O_(2)+LIQ group,the use of purpurin to inhibit PKM2 significantly reversed the trend of changes in the above indicators.Conclusion:LIQ can upregulate PKM2 expression and improve H_(2)O_(2)induced endothelial dysfunction.