脑信号蛋白3A及其受体神经菌毛素-1在慢性肝性脑病大鼠模型中的表达及意义  

Expression and significance of semaphorin 3A and its receptor NRP-1 in a rat model of chronic hepatic encephalopathy

作　　者：阮景晟 陈闯[1] 李科志[1] 黄山[1] 何剑波[1] 曾爱屏[1] 连芳[2] 邬国斌[1] RUAN Jingsheng;CHEN Chuang;LI Kezhi;HUANG Shan;HE Jianbo;ZENG Aiping;LIAN Fang;WU Guobin(Department of Hepatobiliary Surgery,The Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530021,China;College of Basic Medicine,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学附属肿瘤医院肝胆外科,南宁530021 [2]广西医科大学基础医学院,南宁530021

出　　处：《临床肝胆病杂志》2020年第6期1334-1341,共8页Journal of Clinical Hepatology

基　　金：2019年中央引导地方科技发展专项(桂科ZY1949017);广西医药卫生适宜技术推广项目(GZSY20-18);广西高校中青年基础能力提升项目(2018KY0126)。

摘　　要：目的探究肝性脑病(HE)大鼠大脑前额叶皮层、海马CA1、CA3、DG区中脑信号蛋白3A(Sema3A)及受体神经菌毛素(NRP-1)的表达变化及可能的意义。方法SD大鼠40只随机分为4组:对照组、HE 1 d、15 d、30 d组,每组各10只。胆管结扎并辅以乙酸铵灌胃的方法制备慢性HE(C型)大鼠模型,进行行为学、神经病学改变的检测;ELISA检测大鼠血清中血氨及其他生化指标;HE染色观察大鼠肝脏和脑组织形态学变化;Real-time PCR检测各脑区Sema3A、NRP-1 mRNA的表达;免疫组化、Western Blot分别检测大脑皮层、海马CA1、CA3、DG区中Sema3A及NRP-1蛋白的表达变化。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果4组间神经反射等级、血氨、HE等级均有统计学差异(F值分别为76.39、417.07、71.56,P值均<0.001),与对照组比较,HE 1 d、15 d、30 d组神经反射得分、血氨、HE等级均明显升高(P值均<0.05)。4组间TBA、Alb、TBil、AST、ALT、ALP比较差异均有统计学意义(F值分别为1022.61、171.56、685.18、421.01、675.20、2405.04,P值均<0.001),与对照组相比,HE 1 d、15 d、30 d组Alb水平明显降低(P<0.05),其余指标明显升高(P值均<0.05)。病理学结果显示,与对照组相比,造模后大鼠肝脏出现不同程度的变性坏死,炎细胞浸润,大脑前额叶皮层及海马CA1、CA3、DG区体等部位观察到神经细胞不同程度的肿胀、破裂、排列紊乱等病理征象。4组间Sema3A、NRP-1 mRNA比较差异均有统计学意义(Sema3A mRNA:F值分别为273.83、158.84、103.59、128.90,P值均<0.001;mRNA:F值分别为88.78、173.02、156.44、289.09,P值均<0.001);与对照组比较,HE 1 d、15 d、30 d组大脑前额叶皮层、海马CA1、CA3、DG区中Sema3A、NRP-1 mRNA相对表达量均较高(P值均<0.05)。建模后Sema3A、NRP-1蛋白表达水平上调,免疫组化显示,HE组大鼠前额叶皮层和海马CA1、CA3、DG区Sema3A及NRP-1呈阳性染色。Western Blot显�Objective To investigate the changes in the expression of semaphorin 3A(Sema3A)and receptor neuropilins-1(NRP-1)in the prefrontal cortex and the hippocampal CA1,CA3,and DG regions in rats with hepatic encephalopathy(HE)and their possible significance.Methods A total of 40 Sprague-Dawley rats were randomly divided into control group and 1-,15-,and 30-day HE groups,with 10 rats in each group.Bile duct ligation and ammonium acetate by gavage were used to establish a rat model of chronic HE(type C).The behavioral and neurological changes were observed;ELISA was used to measure blood ammonia and related biochemical parameters;HE staining was used to observe morphological changes of the liver and brain tissue;real-time PCR was used to measure the mRNA expression of Sema3A and NRP-1 in each brain region;immunohistochemistry and Western blot were used to measure the protein expression of Sema3A and NRP-1 in the cerebral cortex and the hippocampal CA1,CA3,and DG regions.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results There were significant differences between the four groups in nerve reflex level,blood ammonia,and HE grade(F=76.39,417.07,and 71.56,all P<0.001),and compared with the control group,the 1-,15-,and 30-day HE groups had significant increases in nerve reflex score,blood ammonia,and HE grade(all P<0.05).There were significant differences between the four groups in total bile acid,albumin,total bilirubin,aspartate aminotransferase,alanine aminotransferase,and alkaline phosphatase(F=1022.61,171.56,685.18,421.01,675.98,and 2405.04,all P<0.001),and compared with the control group,the 1-,15-,and 30-day HE groups had a significant reduction in albumin(P<0.05)and significant increases in the other indices(all P<0.05).The pathological results showed that compared with the control group,the HE groups had varying degrees of degeneration,necrosis,and inflammatory cell infil

关 键 词：肝性脑病 信号素3A 神经纤毛蛋白质类 神经元 

分 类 号：R741[医药卫生—神经病学与精神病学] R575[医药卫生—临床医学]