脑蛋白水解物-Ⅰ对缺血再灌注损伤大鼠的神经保护作用  

Neuroprotective effect of cerebroprotein hydrolysate-Ⅰ on ischemia-reperfusion injury in rats

作　　者：翟丽[1] 任宇倩 梁凤[3] 孙浩洋 汪贯习[3] Zhai Li;Ren Yuqian;Liang Feng;Sun Haoyang;Wang Guanxi(Department of Pharmacy,Qingdao Municipal Hospital,Qingdao 266071,China;Center for Integrated Traditional Chinese and Western Medicine,Faculty of Medicine,Qingdao University,Qingdao 266021,China;Department of Radiology,Songshan Hospital,Qingdao University School of Medicine,Qingdao 266021,China)

机构地区：[1]青岛市市立医院药剂科,266071 [2]青岛大学医学部中西医结合中心,青岛266021 [3]青岛大学医学院松山医院放射科,青岛266021

出　　处：《国际脑血管病杂志》2022年第8期589-594,共6页International Journal of Cerebrovascular Diseases

基　　金：青岛大学医疗集团科研专项基金(2020-34)。

摘　　要：目的探讨脑蛋白水解物(cerebroprotein hydrolysate, CH)-Ⅰ对脑缺血再灌注损伤大鼠的神经保护作用及其机制。方法成年健康雄性SD大鼠80只, 随机分为假手术组、模型组、CH-Ⅰ干预组和脑活素(cerebrolysin, CBL)阳性对照组。应用线栓法短暂性闭塞左侧大脑中动脉建立缺血再灌注损伤模型。CH-Ⅰ组和CBL组在再灌注0、3、6和12 h腹腔注射CH-Ⅰ和CBL, 剂量均为20 mg/kg;假手术组和模型组注射同体积生理盐水。在再灌注后24 h, 应用改良神经功能缺损评分(modified Neurological Severity Score, mNSS)检测大鼠行为功能变化, TTC染色检测脑梗死体积, Nissl染色观察缺血皮质区神经细胞形态结构, TUNEL染色检测缺血皮质区神经元凋亡情况, 蛋白质印迹法检测缺血皮质区磷酸化细胞外信号调节激酶(phosphorylated extracellular signal-regulated kinase, pERK)1/2、磷酸化丝裂原活化蛋白激酶激酶(phosphorylated mitogen-activated protein kinase ERK kinase, pMEK)1/2、磷酸化cAMP反应元件结合蛋白(phosphorylated cAMP response element-binding protein, pCREB)和脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)表达的变化。结果再灌注后24 h, 模型组mNSS评分及脑梗死体积显著高于和大于假手术组(P均<0.001);CH-Ⅰ组和CBL组mNSS评分及脑梗死体积均较模型组显著降低(P均<0.05), 但CH-Ⅰ组与CBL组之间差异无统计学意义。Nissl染色和TUNEL染色显示, CH-Ⅰ组变性细胞指数和凋亡细胞指数均显著低于模型组(P均<0.01), 但与CBL组相比差异无统计学意义。蛋白质印迹分析显示, 与假手术组相比, 模型组缺血皮质区pMEK1/2、pERK1/2和pCREB表达显著增强, BDNF表达显著减弱(P<0.05);与模型组比较, CH-Ⅰ组pMEK1/2、pERK1/2和pCREB表达显著降低(P均<0.05), BDNF表达显著增高(P<0.05)。结论 CH-Ⅰ能缩小脑梗死体积和改善神经功能, 其机制可能与抑制MEK-ERK-CREB通路以及增强BDNF表达相关。Objective To investigate the neuroprotective effect of cerebroprotein hydrolysate(CH)-Ⅰon cerebral ischemia-reperfusion injury in rats and its mechanism.Methods Eighty adult healthy male SD rats were randomly divided into sham operation group,model group,CH-Ⅰintervention group and cerebrolysin(CBL)positive control group.The model of ischemia-reperfusion injury was induced by temporarily occluding the left middle cerebral artery with suture-occluded method.The CH-Ⅰand CBL groups intraperitoneally injected with CH-Ⅰand CBL at 0,3,6 and 12 h after reperfusion at the dose of 20 mg/kg.The sham operation group and the model group were injected with the same volume of normal saline.At 24 h after reperfusion,the behavior changes of the rats were detected by the modified neurological severity score(mNSS).The volume of cerebral infarction was detected by TTC staining.The morphology and structure of neurons in ischemic cortex were observed by Nissl staining.The apoptosis of neurons in ischemic cortex was detected by TUNEL staining.The expression changes of phosphorylated extracellular signal-regulated kinase(pERK)1/2,phosphorylated mitogen-activated protein kinase/extracellular signal-regulated kinase(pMEK)1/2,phosphorylated cAMP response element binding protein(pCREB)and brain-derived neurotrophic factor(BDNF)in the ischemic cortex were detected by Western blot.Results At 24 h after reperfusion,the mNSS score and cerebral infarct volume in the model group were significantly higher and larger than those in the sham group(all P<0.001).The mNSS scores and cerebral infarct volumes in the CH-Ⅰand CBL groups were significantly reduced compared with those in the model group(all P<0.05),but there was no significant difference between the CH-Ⅰgroup and the CBL group.Nissl and TUNEL staining showed that the degenerative cell index and apoptotic cell index in the CH-Ⅰgroup were significantly lower than those in the model group(all P<0.01),but there were no significant difference between the CH-Ⅰgroup and the CBL group

关 键 词：脑缺血 再灌注损伤 组织提取物 细胞凋亡 MAP激酶信号系统 神经保护药 疾病模型 动物 大鼠 脑蛋白水解物-I 

分 类 号：R651.15[医药卫生—外科学]