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

阮景晟; 陈闯; 李科志; 黄山; 何剑波; 曾爱屏; 连芳; 邬国斌

doi:10.3969/j.issn.1001-5256.2020.06.029

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

DOI: 10.3969/j.issn.1001-5256.2020.06.029

1. 广西医科大学附属肿瘤医院肝胆外科
2. 广西医科大学基础医学院

基金项目:

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)；

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中图分类号: R575.3;R-332
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出版历程
- 出版日期: 2020-06-20

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

Research funding:

摘要

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、AL...
- 肝性脑病 /
- 信号素3A /
- 神经纤毛蛋白质类 /
- 神经元
Abstract:
Objective To investigate the changes in the expression of semaphorin 3 A( Sema3 A) 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 Sema3 A and NRP-1 in each brain region; immunohistochemistry and Western blot were used to measure the protein expression of Sema3 A 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 infiltration in the liver,with the pathological signs of swelling,tortuous,and ruptured nerve axons in the prefrontal cortex and the hippocampal CA1,CA3,and DG regions. There were significant differences between the four groups in the mRNA expression of Sema3 A and NRP-1( Sema3 A: F = 273. 83,158. 84,103. 59,and 128. 90,all P < 0. 001; NRP-1: F = 88. 78,173. 02,156. 44,and 289. 09,all P<0. 001),and compared with the control group,the 1-,15-,and 30-day HE groups had significantly higher relative mRNA expression of Sema3 A and NRP-1 in the prefrontal cortex and the hippocampal CA1,CA3,and DG regions( all P < 0. 05). There were increases in the protein expression of Sema3 A and NRP-1 after modeling,and immunohistochemistry showed that the HE groups had positive staining of Sema3 A and NRP-1 in the prefrontal cortex and the hippocampal CA1,CA3,and DG regions. Western blot showed that there were significant differences between the four groups in the protein expression of Sema3 A and NRP-1 in the prefrontal cortex and the hippocampal CA1,CA3,and DG regions( Sema3 A: F = 835. 14,774. 16,282. 60,and 856. 29,all P < 0. 001; NRP-1 : F = 876. 59,472. 48,402. 36,and 207. 47,all P < 0. 001),and compared with the control group,the 1-,15-,and 30-day HE groups had significant increases in the protein expression of Sema3 A and NRP-1 in the prefrontal cortex and the hippocampal CA1,CA3,and DG regions( all P < 0. 05). Conclusion Sema3 A/NRP-1 is upregulated in the brain tissue of chronic HE rats and is involved in the pathophysiological process of nerve injury in HE.
- hepatic encephalopathy /
- semaphorin 3A /
- neuropilins /
- neurons

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