羟氯喹促进ERK1/2磷酸化减轻大鼠脑缺血再灌注损伤  

Hydroxychloroquine reduces cerebral ischemia reperfusion injury by promoting ERK1/2 phosphorylation

作　　者：胡秀兰[1] 邵东华[2] 马晓冬[2] 罗红[2] 谢云斌 夏艳 HU Xiu-lan;SHAO Dong-hua;MA Xiao-dong;LUO Hong;XIE Yun-bin;XIA Yan(Department of Intensive Care Unit,,Affiliated Peopled Hospital of Jiangsu University,Zhenjiang Jian-gsu 212002,China;Department of Anesthesiology,Affiliated Peopled Hospital of Jiangsu University,Zhenjiang Jian-gsu 212002,China)

机构地区：[1]江苏大学附属人民医院重症医学科,江苏镇江212002 [2]江苏大学附属人民医院麻醉科,江苏镇江212002

出　　处：《江苏大学学报（医学版）》2019年第1期31-36,共6页Journal of Jiangsu University：Medicine Edition

基　　金：镇江市社会发展项目(SH2015039)

摘　　要：目的:探讨羟氯喹(hydroxychloroquine,HCQ)对大鼠脑缺血再灌注损伤(ischemia/reperfusion,I/R)的影响。方法:采用随机数字表法将健康雄性SD大鼠72只分为6组:对照组、缺血再灌注组(I/R组)、低剂量羟氯喹预处理组(HCQ250组)、中剂量羟氯喹预处理组(HCQ500组)、高剂量羟氯喹预处理组(HCQ1000组)和细胞外信号调节激酶1/2(extracellular regulated protein kinases 1/2,ERK1/2)抑制剂U0126预处理组(U0126组),每组均为12只。采用线栓法行短暂性大鼠大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)建立大鼠缺血性脑卒中模型。I/R组行MCAO后2 h再灌注,HCQ250组、HCQ500组、HCQ1000组和U0126组分别在MCAO前72 h、48 h和24 h经侧脑室注射8μL 250、500、1 000和1 000μmol/L HCQ,其中U0126组在每次注射1 000μmol/L HCQ后12 h再经侧脑室注射8μL 1 000μmol/L U0126;对照组不栓塞大脑中动脉,其余处理同I/R组。再灌注6 h时将每组一半大鼠断头取脑,采用蛋白质印迹法检测缺血半暗带脑组织ERK1/2、p-ERK1/2、抗凋亡因子Bcl-2和促凋亡因子cleaved caspase-3、环磷腺苷效应元件结合蛋白(c AMP-response element binding protein,CREB)、磷酸化CREB(p-CREB)、蛋白激酶B(Akt)和磷酸化Akt(p-Akt)的表达;再灌注24 h时采用神经功能缺陷评分量表检测剩余大鼠的神经功能,再断头取脑后采用2%TTC染色测量脑梗死体积。结果:与对照组比较,其余5组神经功能缺陷评分显著降低,脑梗死体积明显增大,p-ERK1/2、cleaved caspase-3、p-CREB和p-Akt表达显著增高,Bcl-2表达降明显低(P均<0. 05);与I/R组比较,HCQ250组、HCQ500组、HCQ1000组神经功能缺陷评分明显升高,脑梗死体积显著减少,pERK1/2、Bcl-2、p-CREB、p-Akt表达明显增高,cleaved caspase-3表达降低明显(P均<0. 05);与HCQ1000组比较,U0126组神经功能缺陷评分明显降低,脑梗死体积显著增大,p-ERK1/2、Bcl-2、p-CREB和p-Akt表达降低显著,cleaved caspase-3表达明显增高(P均<0. 05)�Objective:To evaluate the attenuation of hydroxychloroquine(HCQ)for cerebral ischemia reperfusion(I/R)injury induced by transient middle cerebral artery occlusion(MCAO)in rats and potential mechanisms.Methods:A total of 72 healthy male SD rats were randomly divided into 6 groups(n=12);control group,ischemia-reperfusion group(group I/R),low dose HCQ pretreatment group(group HCQ250),medium dose HCQ pretreatment group(group HCQ500),high dose HCQ pretreatment group(group HCQ1000)and extracellular regulated protein kinase 1/2(ERK1/2)inhibitor U0126 pretreatment group(group UO126).MCAO was used to establish rat ischemic stroke model.Control group was the same as group I/R,but not occluded the middle cerebral artery;group I/R was reperfused after MCAO for 2 h;group HCQ250,group HCQ500,group HCQ1000 and group U0126 were injected with 8μ250μmol/L,500μmol/L,1 000μmol/L and 1 000μmol/L HCQ by lateral ventricle(LV)at 24 h,48 h and 72 h,respectively,before MCAO,and U0126 group was injected with 8μL of 1 000μmol/L U0126 after 12 h each injection of HCQ.Brain tissues were collected by decollation after reperfusion 6 h,the expression of ERK1/2,p-ERK1/2,anti-apoptotic factor Bcl-2,pro-apoptotic factor cleaved caspase-3,pCREB and p-Akt was detected by Western blotting in ischemic penumbra.Neurological deficit score was measured and followed infarct volume was detection by TTC staining 24 h after reperfusion.Results:Compared with control group,neurological deficit score was reduced,infarct volume was magnified,the expression of p-ERK1/2,cleaved caspase-3,pCREB,pAkt was increased,Bcl-2 expression was decreased in group I/R,group HCQ250,group HCQ500,group HCQ1000 and group U0126(P<0.05).Compared with group I/R,neurological deficit score was enlarged,infarct volume was reduced,the expression of p-ERK1/2,Bcl-2,pCREB,pAkt was increased,the expression of cleaved caspase-3 was decreased in group HCQ1000(P<0.05).However,compared with group HCQ1000,neurological deficit score was reduced,infarct volume was magnified,the expression of

关 键 词：缺血再灌注损伤 羟氯喹 细胞外信号调节激酶 环磷腺苷效应元件结合蛋白 蛋白激酶B 

分 类 号：R743.33[医药卫生—神经病学与精神病学]