葛根素下调Atg5、LC3Ⅰ/Ⅱ蛋白对缺血再灌注脑损伤小鼠的神经保护作用  

Neuroprotective effect of puerarin on ischemic reperfusion brain injury in mice through down-regulating Atg5 and LC3Ⅰ/Ⅱ proteins

作　　者：田华 郭连军[1] 刘富 陈博新 尤梅桂[1] 李楚宜 林虹月 孙裕翔 胡玄桢 TIAN Hua;GUO Lian-jun;LIU Fu;CHEN Bo-xin;YOU Mei-gui;LI Chu-yi;LIN Hong-yue;SUN Yu-xiang;HU Xuan-zhen(Department of Basic Medicine,Xiamen Medical College,Xiamen 361023,Fujian Province,China;Department of Clinical Pharmacology,Qiqihar Medical University,Qiqihar 161006,Heilongjiang Province,China;School of Basic Medicine,Qiqihar Medical University,Qiqihar 161006,Heilongjiang Province,China;Department of Clinical Medicine,Xiamen Medical College,Xiamen 361023,Fujian Province,China)

机构地区：[1]厦门医学院基础医学部,福建厦门361023 [2]齐齐哈尔医学院临床药理教研室,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院基础医学院,黑龙江齐齐哈尔161006 [4]厦门医学院临床医学系,福建厦门361023

出　　处：《中国临床药理学杂志》2023年第4期508-512,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81803757);黑龙江省卫生计生委科研课题基金资助项目(2018471);厦门医学院大学生创新创业基金资助项目(X202112631044)。

摘　　要：目的 观察葛根素通过下调自噬相关基因5(Atg5)、微管相关蛋白轻链Ⅰ/Ⅱ(LC3Ⅰ/Ⅱ)蛋白表达发挥抗缺血再灌注脑损伤小鼠的神经保护作用。方法 将ICR小鼠48只随机分为4组,分别为假手术组、模型组和低剂量、高剂量实验组,每组12只。实验时间共10 d,于第6天夹闭小鼠双侧颈总动脉缺血30 min后进行再灌注,建立缺血性脑损伤小鼠模型,低、高剂量实验组小鼠造模前后5天均腹腔注射葛根素(50,100 mg·kg^(-1)·d^(-1))。用平衡木实验评价小鼠的平衡能力,用自主活动实验评价小鼠的日常活动能力,用蛋白质印迹法检测各组小鼠大脑皮层自噬关键蛋白Atg5、LC3Ⅰ/Ⅱ的相对表达。结果 在术后1 d,假手术组、模型组和低、高剂量实验组小鼠穿越平衡木的时间分别为(5.71±4.23),(21.23±9.49),(10.48±6.50)和(11.33±8.91)s;在术后3 d各组小鼠穿越平衡木的时间分别为(6.09±2.73),(16.83±7.51),(9.48±5.94)和(8.09±5.17)s;在术后5 d各组小鼠穿越平衡木的时间分别为(5.32±2.15),(20.33±11.17),(8.24±5.31)和(8.27±5.54)s;这4组小鼠大脑皮层的LC3Ⅱ蛋白相对表达水平分别为1.00±0.25,1.87±0.14,1.37±0.19和1.03±0.17;这4组小鼠大脑皮层的Atg5蛋白相对表达水平分别为1.00±0.15,1.64±0.38,0.97±0.20和1.01±0.16;上述指标:模型组与假手术组相比,实验组与模型组相比,差异均具有统计学意义(P<0.05或P<0.01)。结论 葛根素可能通过下调自噬关键蛋白Atg5及LC3Ⅰ/Ⅱ发挥抗缺血再灌注脑损伤小鼠的神经保护作用。ObjectiveTo investigate the neuroprotective effect of puerarin against cerebral ischemia reperfusion injury in mice by downregulating autophagy related gene 5 (Atg5) and microtubule associated protein 1 light chainⅠ/Ⅱ(LC3Ⅰ/Ⅱ) protein expression.Methods Forty-eight healthy ICR mice were randomly divided into 4 groups with12 mice in each group,which were sham group,model group and experimental-L,-H groups,respectively.The whole experiment lasted for 10 days.On the sixth day,bilateral common carotid arteries of the mice were occluded for 30 min and then reperfusion was performed to establish a mouse model of ischemic brain injury.The mice in the experimental-L,-H groups were intraperitoneally injected with puerarin 150,100 mg·kg^(-1)5 days before and after modeling.The balance ability of mice was evaluated by balance beam experiment.The daily activity ability of mice was evaluated by autonomous activity experiment.The relative expression of Atg5 and LC3Ⅰ/Ⅱin cerebral cortex of mice in each group was detected by Western blot.Results The time which the mice on 1^(st)day after operation went through balance beam in sham group,model group,experimental-L group and experimental-H group were (5.71±4.23),(21.23±9.49),(10.48±6.50) and (11.33±8.91) s;the time which the mice on 3^(rd)days after operation went through balance beam in these four groups were(6.09±2.73),(16.83±7.51),(9.48±5.94) and(8.09±5.17) s;the time which the mice on 5^(th)day after operation went through balance beam in these four groups were(5.32±2.15),(20.33±11.17),(8.24±5.31) and (8.27±5.54) s,respectively;the relative expression of LC3Ⅱprotein in cerebral cortex of these four groups were 1.00±0.25,1.87±0.14,1.37±0.19 and 1.03±0.17;the relative expression of Atg5 protein in cerebral cortex of these four groups were 1.00±0.15,1.64±0.38,0.97±0.20 and1.01±0.16;compared between the model group and the sham group and compared between the experimental groups and the model group,the differences of the above indicators were stati

关 键 词：缺血再灌注脑损伤 葛根素 平衡能力 自主活动 自噬 

分 类 号：R28[医药卫生—中药学]