国际麻醉学与复苏杂志   2016, Issue (12): 0-0
    
肝硬化门静脉高压对大鼠认知功能和N-甲基-D-天门冬氨酸受体2B亚基表达的影响
谢伟斌1()
1.常州市第三人民医院
Influences of cirrhotic portal hypertension on rats' cognitive function and N-methyl-D-aspartic acid receptor 2B expression level
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摘要:

目的 研究肝硬化门静脉高压对大鼠认知功能及N-甲基-D-天门冬氨酸受体(N-methyl-D-aspartic acid receptor, NR) 2B亚基(NR2B)表达的影响。 方法 雄性SD大鼠37只,按照完全随机法分为两组:肝硬化组(25只),采用四氯化碳-花生油溶液方法制造肝硬化门静脉高压大鼠模型;对照组(12只)。两组于造模前及造模成功后第1、3、5、7天分别利用跳台法和Y-迷宫法对各大鼠进行行为学测试;测试完成后处死大鼠,取海马组织,RT-PCR方法检测大鼠海马NR2B mRNA的水平,Western blot检测海马NR2B蛋白表达水平,免疫组化法检测大鼠海马CA1区NR2B蛋白分布的变化。ELISA法检测肝损伤相关指标ALT、AST、超氧化物歧化酶(superoxide dismutase, SOD)、丙二醛(malondialdehyde, MDA)、TNF-α、IL-1、IL-6的含量。 结果 肝硬化组AST、ALT、TNF-α、IL-1、IL-6、MDA分别为(261±7)、(178±7) U/L、(206±18)、(0.46±0.11)、(298±41) μg/L、(19.7±2.5) μmol/L,较对照组明显升高(P<0.01), SOD为(96±7) U/ml,较对照组明显降低(P<0.01)。与对照组比较:肝硬化组跳台潜伏时间延长、Y-迷宫、错误次数增加(P<0.01),海马中NR2B mRNA的水平及NR2B的蛋白表达明显降低(P<0.01),海马CA1区NR2B免疫组化表达明显下降(P<0.01)。 结论 肝硬化门静脉高压引起大鼠认知功能的降低,同时下调海马中NR2B mRNA和蛋白的表达及CA1区锥形细胞中NR2B的表达。

关键词: 肝硬化; 门静脉高压; 认知功能; N-甲基-D-天门冬氨酸受体2B亚基;
Abstract:

Objective Research on the influences of cirrhotic portal hypertension on rats' hcongnitive function and N-methyl-D-aspartic acid receptor 2B(NR2B) expression level. Methods Thirty seven male SD rats were randomly divided into 2 groups. In the cirrhotic portal hypertension group (group A), 25 rats were inducted into portal hypertension rat model using carbon tetrachloride-peanut oil solution. The remaining 12 rats served as control group (group B). Step-down method and Y-maze method were used in both groups prior to modeling and 1, 3, 5, 7 d after modeling respectively to study their behavior. All rats were killed after testing and hippocampi were collected. RT-PCR was used to detect NR2B mRNA level in the hippocampi, and Western blot was used to detect NR2B protein expression level. Immuno-histochemical method was used to detect NR2B protein distribution changes. ELISA was used to detect the level of ALT and AST, superoxide dismutase (SOD), malondialdehyde (MDA), TNF-α, IL-1 and IL-6. Results The level of AST, ALT, TNF-α, IL-1, IL-6 and MDA in group A were (261±7), (178±7) U/L, (206±18), (0.46±0.11), (298±41) μg/L, (19.7±2.5) μmol/L, which were much higher than in group B(P<0.01), while SOD level[(96.0±7.0) U/ml] was lower than in group B (P<0.01). The step-down latent time of group A was longer and the number of error in Y-maze was more than B group (P<0.01). NR2B mRNA level and NR2B protein expression in the hippocampi of group A rats were evidently lower than that in group B(P<0.01). NR2B immunohistochemical level was lower in the CA1 area of the hippocampi of group A rats than that of the group B (P<0.01). Conclusions Cognitive function were decreased in cirrhotic portal hypertension rats, with decreasing NR2B mRNA and protein expression level in the hippocampus as well. an that of the control group (P<0.01). Conclusions Cognitive function were decreased in cirrhotic portal hypertension rats, by decreasing NR2B expression in the hippocampus.

Key words: Cirrhotic of liver; Portal hypertension; Cognitive function; N-methyl-D-aspartic acid receptor 2B; Rat