目的 基于ERK/Nrf2/HO-1通路探討葡萄籽原花青素（GSP）聯(lián)合亞低溫（MHT）對(duì)腦缺血再灌注損傷大鼠的腦保護(hù)作用。方法 大鼠隨機(jī)分為假手術(shù)組、模型組、MHT組、GSP組、MHT+GSP組。采用線栓法建立腦缺血再灌注損傷大鼠模型。評(píng)估各組大鼠神經(jīng)功能缺損評(píng)分（NDS），TTC法觀察并計(jì)算腦梗死面積，尼氏染色觀察腦組織病理學(xué)變化，TUNEL法計(jì)算細(xì)胞凋亡指數(shù)，酶聯(lián)免疫吸附法檢測(cè)腦組織丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽過(guò)氧化物酶（GSH-Px）水平，Western blotting檢測(cè)Bax、Bcl-2、p-ERK1/2、Nrf2和HO-1蛋白表達(dá)。結(jié)果 與假手術(shù)組比較，模型組大鼠NDS、腦梗死面積、AI、腦組織MDA水平、Bax、p-ERK1/2、胞質(zhì)和胞核Nrf2、HO-1蛋白表達(dá)均顯著升高（P<0.05），SOD、GSH-Px活力和Bcl-2蛋白表達(dá)顯著降低（P<0.05）。與模型組比較，MHT組、GSP組和MHT+GSP組大鼠NDS、腦梗死面積、AI、腦組織MDA水平、Bax和胞質(zhì)Nrf2蛋白顯著降低（P<0.05），SOD、GSH-Px活力、Bcl-2、p-ERK1/2、胞核Nrf2、HO-1蛋白表達(dá)顯著升高（P<0.05）。且MHT+GSP組的治療療效高于MHT組和GSP組。結(jié)論 MHT和GSP聯(lián)合作用可能通過(guò)激活ERK/Nrf2/HO-1通路，上調(diào)p-ERK1/2、胞核Nrf2和HO-1蛋白表達(dá)，抑制氧化應(yīng)激，發(fā)揮腦保護(hù)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

國(guó)家自然科學(xué)基金資助項(xiàng)目（81601078）