基于MAPK信号通路研究脑震宁对三重脑震荡模型大鼠海马神经元损伤的影响  

Effect of Naozhenning on hippocampal neuron injury in multiple cerebral concussion rats model based on MAPK signaling pathway

作　　者：王钱 谭贝西 冯忆 王恬恬 赵乐 王永辉 周然[1] WANG Qian;TAN Bei-xi;FEN Yi;WANG Tian-tian;ZHAO Le;WANG Yong-hui;ZHOU Ran(Hubei University of Chinese Medicine,Wuhan 430065,China;Shanxi University of Chinese Medicine,Jinzhong 030600,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]山西中医药大学,山西晋中030600

出　　处：《中草药》2022年第12期3670-3679,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(8210151374);山西省卫生健康委员会科研项目(2021041);山西中医药大学科技创新能力培育计划“基础研究专项”(2020PY-JC-13)。

摘　　要：目的 研究脑震宁对三重脑震荡模型(multiple cerebral concussion,MCC)大鼠海马神经元损伤的影响及其机制。方法 采用自由落体法制备MCC大鼠模型,将造模成功的大鼠随机分为吡拉西坦(0.16 g/kg)组和脑震宁低、中、高剂量(6.68、13.36、26.73 g/kg)组,各给药组ig相应药物,2次/d,连续给药14 d。造模前后进行水迷宫平台实验,观察大鼠学习记忆能力变化;采用ELISA法检测大鼠血清中白细胞介素-1β(interleukin-1β,IL-1β)、IL-6水平,WST-1法检测血清超氧化物歧化酶(superoxide dismutase,SOD)活性,硫代巴比妥酸法检测血清丙二醛(malondialdehyde,MDA)水平;qRTPCR法检测大鼠海马组织p38、细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)、c-Jun氨基末端激酶(cJun N-terminal kinase,JNK)m RNA表达;Western blotting法检测大鼠海马组织p38、p-p38、ERK、p-ERK、JNK和p-JNK蛋白表达;采用苏木素-伊红(HE)、尼氏体染色以及透射电镜(TEM)法分别观察大鼠海马神经元病理、尼氏体及超微结构的变化。结果 与对照组相比,模型组大鼠逃避潜伏时间增加(P<0.01),固定象限停留时间百分比减少(P<0.01);血清中IL-1β、IL-6和MDA水平升高(P<0.05、0.01),SOD活性下降(P<0.01);海马组织p38、JNK和ERK mRNA表达水平均升高(P<0.01);海马组织p38、p-p38、JNK、p-JNK、ERK和p-ERK蛋白表达水平均升高(P<0.05、0.01)。与模型组相比,脑震宁组大鼠逃避潜伏时间减少(P<0.05、0.01),固定象限停留时间百分比增加(P<0.05、0.01);血清中IL-1β、IL-6、MDA水平降低(P<0.05、0.01),SOD活性升高(P<0.01、0.001);海马组织p38、JNK和ERK m RNA表达水平降低(P<0.05、0.01);脑震宁各剂量组海马组织p38、p-p38和p-ERK蛋白表达水平降低(P<0.01),脑震宁低、中剂量组JNK和ERK蛋白表达水平降低(P<0.05),脑震宁中、高剂量组p-JNK蛋白表达水平升高(P<0.01)。HE、尼氏染色结果显示,模型组神经元排列稀疏,数量减少,部分细胞�Objective To study the effect and mechanism of Naozhenning(脑震宁)on hippocampal neuron injury in multiple cerebral concussion(MCC)model rats.Methods Free fall method was processed to establish MCC models,MCC rats were randomly divided into piracetam(0.16 g/kg)group and low-,medium-,high-dose Naozhenning(6.68,13.36,26.73 g/kg)groups,each administration group was ig given corresponding drugs,twice a day for 14 d.Water maze platform experiment was carried out before and after modeling to observe the changes of learning and memory ability of rats;Levels of interleukin-1β(IL-1β)and IL-6 in serum of rats were detected by ELISA;Superoxide dismutase(SOD)content was detected by WST-1 method;Malondialdehyde(MDA)content was detected by thiobarbituric acid method;mRNA expressions of p38,extracellular regulated protein kinases(ERK)and c-Jun N-terminal kinase(JNK)in hippocampus were detected by qRT-PCR;p38,p-p38,ERK,p-ERK,JNK and p-JNK protein expressions in hippocampus were detected by Western blotting;HE,Nissl staining and transmission electron microscopy were used to observe the changes of pathology,Nissl body and ultrastructure of hippocampal neurons.Results Compared with control group,escape latency in model group was increased(P<0.01),percentage of residence time in fixed quadrant was decreased(P<0.01);IL-1β,IL-6 and MDA levels in serum were increased(P<0.05,0.01),SOD activity was decreased(P<0.01);mRNA expressions of p38,JNK and ERK in hippocampus were increased(P<0.01);p38,p-p38,JNK,p-JNK,ERK and p-ERK protein expressions in hippocampus were increased(P<0.05,0.01).Compared with model group,escape latency of rats in Naozhenning groups was decreased(P<0.05,0.01),percentage of fixed quadrant residence time was increased(P<0.05,0.01);IL-1β,IL-6 and MDA levels were decreased(P<0.05,0.01),SOD activity was increased(P<0.01,0.001);mRNA expressions of p38,JNK and ERK in hippocampus were decreased(P<0.05,0.01);p38,p-p38 and p-ERK protein expressions of hippocampus in Naozhenning groups were decreased(P<0.01),JNK and ERK

关 键 词：脑震宁 丝裂原活化蛋白激酶 神经元 三重脑震荡模型 炎症 氧化应激 

分 类 号：R285.5[医药卫生—中药学]