氟比洛芬通过PI3K/AKT/PHB2通路抑制线粒体诱导的心肌细胞氧化应激损伤  

Study on the mechanism of Flurbiprofen on cardiomyocyte injury induced by oxidative stress

作　　者：张熙 逄晓倩 苗雨欣 贡美娜 田炜 徐菁蔓 ZHANG Xi;PANG Xiaoqian;MIAO Yuxin;GONG Meina;TIAN Wei;XU Jingman(School of public health,North China University of Science and Technology,Tangshan 063210,China)

机构地区：[1]华北理工大学公共卫生学院,河北省唐山市063210 [2]河北省器官纤维化重点实验室 [3]华北理工大学实验动物中心

出　　处：《中国煤炭工业医学杂志》2024年第3期225-232,共8页Chinese Journal of Coal Industry Medicine

基　　金：中央引导地方科技发展资金项目(编号:226Z7711G);河北省引进留学人员资助项目(编号:C20220354);华北理工大学公共卫生学院青年人才托举计划(编号:QNRC202313)。

摘　　要：目的探讨氟比洛芬能否减缓氧化应激诱发心肌细胞损伤及其可能机制。方法建立心肌H9c2细胞氧化应激损伤模型并分为对照(Control)组、过氧化氢(H_(2)O_(2))组、氟比洛芬+过氧化氢(F+H_(2)O_(2))组、氟比洛芬(F)组、氟比洛芬+过氧化氢+渥曼青霉素(F+H_(2)O_(2)+Wort)组、渥曼青霉素(Wort)组。MTT法检测细胞活性;激光共聚焦显微镜检测线粒体膜电位的变化;利用Western blot技术检测磷酸化糖原合成酶激酶-3β(p-GSK-3β),磷酸化蛋白激酶-B(p-AKT)抗增殖蛋白1(PHB1),抗增殖蛋白2(PHB2)的表达水平。结果与H_(2)O_(2)组比较,F+H_(2)O_(2)(10、50、100μmol/L)组细胞存活率升高,差异具有统计学意义(P<0.05)。与H_(2)O_(2)组比较,F+H_(2)O_(2)组中的线粒体膜电位水平升高且差异具有统计学意义(P<0.05);与F+H_(2)O_(2)组相比,F+H_(2)O_(2)+Wort组的线粒体膜电位水平下降,差异具有统计学意义(P<0.05)。与H_(2)O_(2)组相比,F+H_(2)O_(2)组中p-GSK-3β、p-AKT、PHB2蛋白表达水平提高,差异具有统计学意义(P<0.05);但F+H_(2)O_(2)组中PHB1蛋白表达水平无变化(P>0.05)。与F+H_(2)O_(2)组比较,F+H_(2)O_(2)+Wort组中的PHB2、p-GSK-3β、p-AKT蛋白表达水平下降,差异具有统计学意义(P<0.05);但F+H_(2)O_(2)+Wort组中PHB1蛋白表达水平无变化(P>0.05)。结论氟比洛芬可能通过激活PI3K/AKT信号途径,一方面抑制GSK-3β活性,另一方面增加PHB2,进而抑制线粒体膜通透性转换孔(mPTP)开放,减轻氧化应激引起的心肌H9c2细胞损伤。Objective To investigate whether flurbiprofen can reduce oxidative stress-induced cardiomyocyte injury and its specific mechanism.Methods The oxidative stress injury model of myocardial H9c2 cells was established and divided into Control group,H_(2)O_(2)group,F+H_(2)O_(2)group,F group,F+H_(2)O_(2)+Wort group,and Wort group.The cell activity was measured by MTT.The expression of anti-proliferating proteins PHB1 and PHB2,phosphorylated glycogen synthetase kinase-3β(p-GSK-3β)and phosphorylated protein kinase-B(p-AKT)were detected by Western blot.Using laser confocal microscopy to observe mitochondrial membrane potential.Results Compared with the H_(2)O_(2)group,the F+H_(2)O_(2)(10,50,100μmol/L)group showed a statistically significant difference(P<0.05).Compared with the H_(2)O_(2)group,the difference of mitochondrial membrane potential level in the F+H_(2)O_(2)group was statistically significant(P<0.05);the decrease in the mitochondrial membrane potential level in the F+H_(2)O_(2)+Wort group compared with the F+H_(2)O_(2)group was statistically significant(P<0.05).Compared with the H_(2)O_(2)group,p-GSK-3β,p-AKT and PHB2 were increased in the F+H_(2)O_(2)group(P<0.05);but PHB1 was unchanged in the F+H_(2)O_(2)group(P>0.05).Compared with the F+H_(2)O_(2)group,PHB2,p-GSK-3β,and p-AKT proteins in the F+H_(2)O_(2)+Wort group were statistically significant(P<0.05);however,PHB1 protein was unchanged in the F+H_(2)O_(2)+Wort group(P>0.05).Conclusion Flurbiprofen may alleviate cardiac H9c2 cell oxidative damage induced mitochondrial permeablity transition pore(mPTP)opening by inhibiting GSK-3βand increasing PHB2 via activating PI3K/AKT signaling pathway.

关 键 词：氟比洛芬 氧化应激损伤 PHB PI3K/AKT GSK-3Β 

分 类 号：R364.5[医药卫生—病理学]