熊果酸减轻高糖高脂诱导的人主动脉内皮细胞损伤

Ursolic acid alleviates the injury of human aortic endothelial cells induced by high glucose and high fat

目的:探讨熊果酸(ursolic acid,UA)对人主动脉内皮细胞(human aortic endothelial cells,HAEC)高糖高脂损伤的影响及机制。方法:MTT法选取合适的高糖和棕榈酸钠(sodium palmitate,SPA)损伤浓度以及UA预孵浓度。采用试剂盒检测NO、ROS的含量,乳酸脱氢酶(LDH)的释放率以及抗氧化酶包括总超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性。Western blot法检测内皮型一氧化氮合酶(eNOS)、细胞间黏附分子(ICAM-1)、血管细胞黏附分子(VCAM-1)、半胱天冬酶-1(caspase-1)以及GSDMD的蛋白表达情况。Hoechst33342联合PI荧光染色检测细胞膜的完整状态以探讨焦亡发生情况。观察UA对HAEC的保护作用。结果:预孵UA(1μmol/L和5μmol/L)可以减轻高糖高脂(30 mmol/L GLU+0.1 mmol/L SPA)造成的HAEC损伤,提升HAEC活力,减轻氧化应激损伤,增加NO的释放和eNOS的蛋白表达,减少黏附分子的蛋白表达,减少细胞焦亡。结论:UA可能通过减轻高糖高脂诱导的HAEC氧化应激反应,抑制细胞焦亡的发生,减轻内皮细胞损伤。

AIM:To investigate the effect and mechanism of ursolic acid on high glucose and high-fat injury of human aortic endothelial cells.METHODS:MTT method was used to select the appropriate injury concentration of high glucose and sodium palmitate and UA pre incubation concentration.The levels of NO and ROS,the release rate of lactate dehydrogenase and the activities of antioxidant enzymes including total superoxide dismutase,catalase and glutathione peroxidase were detected by kit.The expression of endothelial nitric oxide synthase,intercellular adhesion molecule,vascular cell adhesion molecule caspase-1 and GSDMD were detected by Western blot.The protective effect of UA on HAEC was observed.Hoechst33342 combined with PI fluorescence staining was used to detect the whole state of cell membrane to explore the occurrence of pyroptosis.RESULTS:Pre-incubation with UA(1 and 5μmol/L)could reduce the damage of HAEC caused by high glucose and high fat(30 mmol/L Glu+0.1 mmol/L SPA),enhance HAEC activity,increase NO release and eNOS protein expression,alleviate oxidative stress injury,reduce the protein expression of adhesion molecules and reduce the occurrence of pyroptosis.CONCLUSION:UA may reduce the damage of endothelial cells by inhibiting the oxidative stress response and the occurrence of pyroptosis induced by high glucose and high fat.