二甲双胍对局灶性脑缺血再灌注损伤大鼠神经保护作用机制研究  

Neuroprotective mechanism of metformin in rats with focal cerebral ischemia-reperfusion injury

作　　者：刘磊 冯禧轩[1] 马宇莘 王璐 LIU Lei;FENG Xi-xuan;MA Yu-xin(Department of Human Anatomy and Histology,Yan'an University,Yan'an Shaanxi 716000,China;Department of Urology,Yan'an People's Hospital,Yan'an Shaanxi 716000,China.)

机构地区：[1]延安大学人体解剖与组织胚胎学教研室,陕西延安716000 [2]延安市人民医院泌尿外科,陕西延安716000

出　　处：《临床和实验医学杂志》2024年第20期2129-2133,共5页Journal of Clinical and Experimental Medicine

基　　金：陕西省科技厅立项(编号:2021SF-082)。

摘　　要：目的探讨二甲双胍对局灶性脑缺血再灌注损伤大鼠神经保护作用的机制。方法选择30只8周龄健康雄性SPF级SD大鼠为研究对象,按随机数字表法分为假手术组、大脑中动脉闭塞(MCAO)组及二甲双胍组(MCAO+Met组),每组各10只。在构建大鼠MCAO模型前3周,MCAO+Met组行10 mg/kg二甲双胍腹腔注射,假手术组及MCAO组不作任何药物处理,随后参考Longa线栓法制备大鼠MCAO模型,假手术组除不插入线栓,余下操作同MCAO组、MCAO+Met组。依据造模成功标准,在造模成功24 h后采用改良神经功能评分(mNSS)评定神经功能缺损程度;采用氯化三苯基四氮唑(TTC)染色测定大鼠脑梗死体积;采用蛋白质印迹法测定磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(AKT)及血管内皮生长因子(VEGF)蛋白的表达水平;采用酶联免疫吸附试验测定白细胞介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)水平。比较3组大鼠mNSS评分、脑梗死体积百分数、PI3K、AKT、VEGF蛋白表达、IL-6、IL-1β及TNF-α水平。结果与假手术组比较,MCAO组及MCAO+Met组mNSS评分均更高,差异均有统计学意义(P<0.05);与MCAO组比较,MCAO+Met组mNSS评分更低,差异有统计学意义(P<0.05)。与假手术组比较,MCAO组及MCAO+Met组脑梗死体积百分数均更高,差异均有统计学意义(P<0.05);与MCAO组比较,MCAO+Met组脑梗死体积百分数更低,差异有统计学意义(P<0.05)。与假手术组比较,MCAO组及MCAO+Met组PI3K、AKT及VEGF蛋白表达水平均更低,差异均有统计学意义(P<0.05);与MCAO组比较,MCAO+Met组脑梗死体积PI3K、AKT及VEGF蛋白表达水平均更高,差异均有统计学意义(P<0.05)。与假手术组比较,MCAO组及MCAO+Met组IL-6、IL-1β及TNF-α水平均更高,差异均有统计学意义(P<0.05);与MCAO组比较,MCAO+Met组IL-6、IL-1β及TNF-α水平均更低,差异均有统计学意义(P<0.05)。结论局灶性脑缺血再灌注损伤应用二甲双胍有利于神经保护,其作用机制可能与激活PI3KObjective To investigate the neuroprotective mechanism of metformin in rats with focal cerebral ischemia-reperfusion injury.Methods Thirty healthy 8-week-old male SPF SD rats were divided into Sham group,middle cerebral artery occlusion(MCAO)group and metformin group(MCAO+Met group)according to random number table method,with 10 in each group.The MCAO+Met group was given intrabitoneal injection of 10 mg/kg Met 3 weeks before the establishment of the rat MCAO model.The Sham group and MCAO group were not treated with any drug,and then the rat model of MCAO was prepared by Longa method.The remaining operations in Sham group were the same as those in MCAO group and MCAO+Met group except that no plug was inserted.According to the success criteria of modeling,the degree of neurological impairment was assessed by modified neurological severity score(mNSS),the cerebral infarction volume was determined by triphenyltetrazolium chloride(TTC)staining,and the expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT)and vascular endothelial growth factor(VEGF)proteins were determined by Western blotting.The levels of interleukin(IL),IL-1β,and tumor necrosis factor-α(TNF-α)were measured by enzyme linked immunosorbent assay.The mNSS score,percentage of cerebral infarction volume,PI3K,AKT,VEGF protein expression,IL-6,IL-1βand TNF-αlevels were compared among the three groups.Results Compared with Sham group,mNSS score in MCAO group and MCAO+Met group were higher,the differences were statistically significant(P<0.05);mNSS score in MCAO+Met group were lower than that in MCAO group,the difference was statistically significant(P<0.05).Compared with Sham group,the volume of cerebral infarction in MCAO group and MCAO+Met group were higher,the difference were statistically significant(P<0.05);compared with MCAO group,the volume of cerebral infarction in MCAO+Met group was lower,the difference was statistically significant(P<0.05).Compared with Sham group,the expression levels of PI3K,AKT and VEGF protein in MCAO

关 键 词：大鼠 二甲双胍 局灶性脑缺血再灌注损伤 作用机制 

分 类 号：R741[医药卫生—神经病学与精神病学]