对羟基苯甲醇通过SIRT1/PGC-1α/TFAM信号通路抗脑缺血再灌注损伤的作用及机制研究  

Effect and mechanism of p-hydroxybenzyl alcohol on cerebral ischemia-reperfusion injury by SIRT1/PGC-1α/TFAM signaling pathway

作　　者：余星霖 罗苑 杨丽萍 陈普 段小花 YU Xing-lin;LUO Yuan;YANG Li-ping;CHEN Pu;DUAN Xiao-hua(Yunnan Key Laboratory for Dai and Yi Medicines,Yunnan University of Chinese Medicine,Yunnan Kunming 650500,China)

机构地区：[1]云南中医药大学,云南省傣医药与彝医药重点实验室,云南昆明650500

出　　处：《中国医院药学杂志》2023年第1期42-47,共6页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目(编号:81960733);云南省科技厅基础研究面上项目(编号:2019FB120)。

摘　　要：目的:基于SIRT1/PGC-1α/TFAM信号通路探讨对羟基苯甲醇(HBA)抗脑缺血再灌注损伤作用。方法:通过线栓法复制MCAO/R大鼠模型,分为假手术组、模型组(MCAO/R)、HBA组、抑制剂(EX527)组。其中抑制剂组是侧脑注射EX527(10 mmol·L^(-1),10μL),HBA治疗组给予HBA 20 mg·kg^(-1)进行药物干预。旷场实验、转棒实验、Morris水迷宫实验观察大鼠行为学变化;TTC法检测脑梗死体积;取缺血侧脑组织,电镜观察线粒体显微结构变化,检测组织中活性氧(ROS)、线粒体膜电位(MMP)、呼吸链酶复合体Ⅰ~Ⅳ活性、ATP水平;免疫组化法检测脑源性神经营养因子(BDNF)和神经生长相关蛋白(GAP-43)蛋白阳性表达;Western blot法检测脑组织中SIRT1、PGC-1a、NRF1/2、TFAM蛋白表达。结果:HBA干预后能改善MCAO/R大鼠记忆空间学习及运动能力等行为学变化,明显降低脑梗死体积;改善线粒体结构损伤,降低ROS水平,提高线粒体呼吸链酶复合体Ⅰ~Ⅳ活性、MMP以及ATP水平。促进BDNF和GAP-43及SIRT1/PGC-1a途径相关蛋白表达。SIRT1抑制剂处理后可逆转HBA的治疗作用。结论:HBA可促进缺血再灌注过程中神经元的修复,其可能与激活SIRT1/PGC-1a/TFAM信号通路有关。OBJECTIVE To investigate the effect of p-hydroxybenzyl alcohol(HBA) on cerebral ischemia-reperfusion injury(CIRI) based on SIRT1/PGC-1α/TFAM signaling pathway.METHODS MCAO/R rats were divided into sham operation group, model group(MCAO/R), HBA group and inhibitor group(EX527). The open field test, rotary rod test and Morris water maze test were used to observe the behavioral changes of rats. The volume of cerebral infarction was detected by TTC. The ischemic brain tissues were collected to observe the changes of mitochondrial microstructure, and the reactive oxygen species(ROS), mitochondrial membrane potential(MMP), respiratory chain enzyme complex Ⅰ-Ⅳ activities and ATP levels were detected by electron microscopy. The expression of brain derived neurotrophic factor(BDNF) and nerve growth associated protein(GAP-43) was detected by immunohistochemistry. The protein expression of SIRT1, PGC-1a, NRF1/2 and TFAM in brain tissue was detected by Western blot.RESULTS HBA intervention could improve the behavioral changes of MCAO/R rats, such as memory, spatial learning and motor ability, and significantly reduce the volume of cerebral infarction. In addition, the damage of mitochondrial structure was ameliorated, ROS levels were decreased, mitochondrial respiratory chain enzyme complex Ⅰ activity, MMP and ATP levels were increased. The expression of BDNF, GAP-43 and SIRT1/PGC-1a pathway related proteins was promoted. Treatment with SIRT1 inhibitor could reverse the therapeutic effects of HBA.CONCLUSION HBA can promote neuronal repair during ischemia-reperfusion, which may be related to the activation of SIRT1/PGC-1a/TFAM signaling pathway by HBA.

关 键 词：对羟基苯甲醇 MCAO/R大鼠 SIRT1/PGC-1α/TFAM信号通路 线粒体 

分 类 号：R285.5[医药卫生—中药学]