冰片对过氧化氢损伤后大鼠心肌微血管内皮细胞的保护作用

Protective Effect of Borneol on Rat Cardiac Microvascular Endothelial Cells after Hydrogen Peroxide Injury

目的研究冰片对心肌微血管内皮细胞的保护作用。方法体外培养的大鼠心肌微血管内皮细胞(cardiac microvascular endothelial cells,CMECs),采用CCK-8(cell counting kit-8)检测细胞活力,建立过氧化氢H_(2)O_(2)损伤CMECs模型,确定实验适宜的H_(2)O_(2)浓度及冰片治疗浓度。将CMECs分为对照组、H_(2)O_(2)损伤组及冰片治疗组。通过检测氧化应激反应多种中间产物含量及线粒体功能等,观察比较冰片对H_(2)O_(2)诱导的CMECs损伤的保护作用。结果H_(2)O_(2)损伤组与对照组比较,乳酸脱氢酶(lactate dehydrogenase,LDH)、丙二醛(malondialdehyde,MDA)、NADPH氧化酶、氧化型谷胱甘肽(oxidized glutathione,GSSG)含量明显增加,还原型谷胱甘肽(reduced glutathione,GSH)含量明显减少;经冰片治疗后,CMECs内LDH、MDA、NADPH氧化酶、GSSG含量明显减少,GSH含量明显增加。与对照组比较,H_(2)O_(2)损伤组FACL4基因mRNA及蛋白的表达显著升高,GPX4基因mRNA及蛋白的表达显著降低;冰片治疗组与H_(2)O_(2)损伤组比较,FACL4基因mRNA及蛋白的表达降低,GPX4基因mRNA及蛋白的表达增加。结论冰片对H_(2)O_(2)诱导的CMECs氧化应激损伤及线粒体生物合成功能的损伤具有保护作用。

Objective To study the protective effect of Borneol,one of the effective ingredients of Shexiang Baoxin Pill,on cardiac microvascular endothelial cells.Methods Rat cardiac microvascular endothelial cells(CMECs)cultured in vitro were used to detect cell viability by CCK-8(cell counting kit-8)to establish a model of hydrogen peroxide damage to CMECs,and determine the appropriate H_(2)O_(2) concentration for the experiment and the therapeutic concentration of borneol.The CMECs were divided into control group,H_(2)O_(2) injury group and borneol treatment group.By detecting the content of various intermediate products of oxidative stress reaction and the function of Mitochondria,we observed and compared the protective effects of borneol on H_(2)O_(2)-induced damage of CMECs.Results Compared with the control group,the content of lactate dehydrogenase(LDH),malondialdehyde(MDA),NADPH oxidase,and oxidized glutathione(GSSG)in the H_(2)O_(2) injury group were significantly increased,and the content of reduced glutathione(GSH)was significantly reduced.After treatment,the content of LDH,MDA,NADPH oxidase,and GSSG in CMECs were significantly reduced,and the content of GSH was significantly increased.Compared with the control group,the expression of FACL4mRNA and protein in the H_(2)O_(2) injury group was significantly increased,and the expression of GPX4mRNA and protein was significantly reduced;compared with the H_(2)O_(2) injury group,the expression of FACL4mRNA and protein was reduced,and the expression of GPX4mRNA and protein increased in the borneol group.Conclusion Borneol can protect CMECs from oxidative stress damage and mitochondrial biosynthesis damage induced by H_(2)O_(2).