通过内质网应激和 MAPK 介导的机制保护大脑免受缺血/再灌注损伤。

protects against cerebral ischemia/reperfusion injury through endoplasmic reticulum stress and MAPK-mediated mechanisms.

机构信息

School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China.

Jiangsu Kanion Parmaceutical Co. Ltd., Lianyungang, Jiangsu 222001, China.

出版信息

J Adv Res. 2021 Feb 9;33:215-225. doi: 10.1016/j.jare.2021.01.016. eCollection 2021 Nov.

DOI:10.1016/j.jare.2021.01.016

PMID:34603791

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8463917/

Abstract

INTRODUCTION

(LTC) is wildly applied to treat ischemic stroke in clinical practice in China. However, the pharmacological mechanism of LTC on ischemic stroke is still unstated.

OBJECTIVE

Our research was designed to study the protective effect of LTC against cerebral ischemia-reperfusion (I/R) injury and reveal the underlying mechanism both .

METHODS

PC12 cells treated with glucose deprivation/reperfusion (OGD/R) were used to simulate ischemia/reperfusion (I/R) injury. The cell viability, apoptosis rate, and protein expressions of PC12 cells were evaluated. validation of the protective effect of LTC was carried out by middle cerebral artery occlusion (MCAO)/reperfusion treatment, and the underlying mechanism of its anti-apoptosis ability was further revealed by immunohistochemistry staining and Western blotting.

RESULTS

In the current study, we observed that LTC effectively inhibited oxygen-glucose deprivation/reperfusion (OGD/R) induced apoptosis of PC12 cells through suppressing the cleavage of poly ADP-ribose polymerase (PARP), caspase-3, and caspase-9. Further investigation revealed that OGD/R insult remarkably triggered the endoplasmic reticulum stress responses (ER stress) to induce PC12 cell apoptosis. LTC treatment alleviated OGD/R induced ER stress by inhibiting the activation of protein kinase RNA (PKR)-like ER kinase (PERK)/eukaryotic translation initiation factor 2 (eIF2α) and inositol requiring enzyme 1 (IRE1)/tumor necrosis factor receptor-associated factor 2 (TRAF2) pathways. Additionally, LTC also restrained the OGD/R-induced PC12 cell apoptosis by reversing the activated mitogen-activated protein kinase (MAPK) through IRE1/TRAF2 pathway. Animal studies demonstrated LTC significantly restricted the infarct region induced by middle cerebral artery occlusion (MCAO)/reperfusion, the activation of ER stress and apoptosis of neuronal cells had also been suppressed by LTC in the penumbra region.

CONCLUSION

LTC protects the cerebral neuronal cell against ischemia/reperfusion injury through ER stress and MAPK-mediated mechanisms.

摘要

简介

（LTC）在中国临床上广泛应用于治疗缺血性中风。然而，LTC 对缺血性中风的药理机制尚不清楚。

目的

本研究旨在研究 LTC 对脑缺血再灌注（I/R）损伤的保护作用，并揭示其潜在机制。

方法

用葡萄糖剥夺/再灌注（OGD/R）处理 PC12 细胞，模拟缺血/再灌注（I/R）损伤。评估 PC12 细胞的细胞活力、细胞凋亡率和蛋白表达。通过大脑中动脉闭塞（MCAO）/再灌注处理验证 LTC 的保护作用，并通过免疫组织化学染色和 Western blot 进一步揭示其抗细胞凋亡能力的潜在机制。

结果

在本研究中，我们观察到 LTC 通过抑制多聚 ADP-核糖聚合酶（PARP）、半胱天冬酶-3 和半胱天冬酶-9 的裂解，有效抑制氧-葡萄糖剥夺/再灌注（OGD/R）诱导的 PC12 细胞凋亡。进一步研究表明，OGD/R 损伤可显著引发内质网应激反应（ER 应激），诱导 PC12 细胞凋亡。LTC 通过抑制蛋白激酶 RNA（PKR）样内质网激酶（PERK）/真核翻译起始因子 2（eIF2α）和肌醇需要酶 1（IRE1）/肿瘤坏死因子受体相关因子 2（TRAF2）途径，减轻 OGD/R 诱导的 ER 应激。此外，LTC 还通过 IRE1/TRAF2 途径逆转激活的丝裂原活化蛋白激酶（MAPK），抑制 OGD/R 诱导的 PC12 细胞凋亡。动物研究表明，LTC 显著限制了大脑中动脉闭塞（MCAO）/再灌注引起的梗死区，同时 LTC 还抑制了缺血半影区的 ER 应激和神经元细胞的凋亡。