目的 基于ERK/Nrf2/HO-1通路探讨葡萄籽原花青素（GSP）联合亚低温（MHT）对脑缺血再灌注损伤大鼠的脑保护作用。方法 大鼠随机分为假手术组、模型组、MHT组、GSP组、MHT+GSP组。采用线栓法建立脑缺血再灌注损伤大鼠模型。评估各组大鼠神经功能缺损评分（NDS），TTC法观察并计算脑梗死面积，尼氏染色观察脑组织病理学变化，TUNEL法计算细胞凋亡指数，酶联免疫吸附法检测脑组织丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）水平，Western blotting检测Bax、Bcl-2、p-ERK1/2、Nrf2和HO-1蛋白表达。结果 与假手术组比较，模型组大鼠NDS、脑梗死面积、AI、脑组织MDA水平、Bax、p-ERK1/2、胞质和胞核Nrf2、HO-1蛋白表达均显著升高（P<0.05），SOD、GSH-Px活力和Bcl-2蛋白表达显著降低（P<0.05）。与模型组比较，MHT组、GSP组和MHT+GSP组大鼠NDS、脑梗死面积、AI、脑组织MDA水平、Bax和胞质Nrf2蛋白显著降低（P<0.05），SOD、GSH-Px活力、Bcl-2、p-ERK1/2、胞核Nrf2、HO-1蛋白表达显著升高（P<0.05）。且MHT+GSP组的治疗疗效高于MHT组和GSP组。结论 MHT和GSP联合作用可能通过激活ERK/Nrf2/HO-1通路，上调p-ERK1/2、胞核Nrf2和HO-1蛋白表达，抑制氧化应激，发挥脑保护作用。

Objective To explore the protective effect of grape seed proanthocyanidins (GSP) combined with mild hypothermia (MHT)on cerebral ischemia-reperfusion injury in rats based on ERK/Nrf2/HO-1 pathway. Methods Mice were randomly divided into sham operation group, model group, MHT group, GSP group, and MHT + GSP group. Mice with cerebral ischemia-reperfusion injury were established by thread embolization. The neurological deficit score (NDS) of rats in each group were evaluated, and the cerebral infarction area observed and calculated by TTC method. The brain histopathological changes were observed by Nissl staining, and apoptotic index was calculated by TUNEL method. The MDA, SOD, and GSH-Px in brain tissues were detected by enzyme-linked immunosorbent assay, and Bax, Bcl-2, p-ERK1/2, Nrf 2, and HO-1 protein expression were measured by Western blotting analysis. Results Compared with sham-operated group, NDS, cerebral infarction area, AI, and MDA content in brain tissue, Bax, p-ERK1/2, Nrf2, and HO-1 protein expression in cytoplasm and nucleus were significantly increased (P<0.05), but SOD, GSH-Px activity, Bcl-2 protein expression were significantly decreased (P<0.05). Compared with model group, NDS, cerebral infarction area, AI and MDA content in brain tissue, Bax, and cytoplasmic Nrf2 protein were significantly decreased in MHT group, GSP group and MHT+GSP group (P<0.05), but Bcl-2, p-ERK1/2, Nrf2 and HO-1 protein expression in nucleus were significantly increased (P<0.05). And the therapeutic effect of MHT + GSP group was higher than that of MHT group and GSP group. Conclusion The combined effect of MHT and GSP may play a protective role in brain by activating ERK/Nrf2/HO-1 pathway, up-regulating the expression of p-ERK1/2, Nrf2 and HO-1 protein in nucleus, inhibiting oxidative stress.

R966;R285.5

国家自然科学基金资助项目（81601078）