iNOS、nNOS在急性一氧化碳中毒迟发脑病中的表达及其与神经元变性坏死的相关性研究

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目的:检测iNOS、nNOS在急性CO中毒迟发性脑病中表达及其与海马神经元变性坏死的相关性分析,探索iNOS、nNOS在急性一氧化碳中毒迟发性脑病(delayed encephalopathy after acute carbon monoxide poisoning, DEACMP)发病中的作用。方法:选择成年雄性SD大鼠72只,随机(随机数字法)分为DEACMP组、正常组,每组36只,分别给予腹腔注射99.99%的CO气体、腹腔不干预。再根据染造模前后不同时间段,将两组再分为造模前、造模1 d、造模7 d、造模14 d、造模21 d、造模28 d的6个亚组,在6个亚组的时间段上,行HE染色观察海马CA3区神经元变性坏死,行免疫组织化学、Western blot方法检测海马区iNOS、nNOS的蛋白表达。应用SPSS 21.0软件统计分析。计量资料用(Mean±SD)表示,对各组实验结果进行正态性检验及方差分析,各组间均数比较采用两独立样本的n t检验,iNOS、nNOS蛋白相对表达量与变性坏死神经元进行相关性分析,符合正态性分布用Pearson相关分析,不符合正态性分布用Spearman秩相关分析,以n P0.05),在造模14 d、造模21 d、造模28 d两组比较差异具差异有统计学意义(n P0.05),在造模1 d、造模7 d、造模14 d比较差异具差异有统计学意义(n P0.05),在造模7 d、造模14 d、造模21 d、造模28 d比较差异具差异有统计学意义(n P<0.05)。iNOS蛋白表达与变性坏死神经元计数之间进行相关性分析,两者之间呈现正相关(n P0.05)。n 结论:iNOS在DEACMP的发病过程中起了重要作用;而nNOS在DEACMP海马中没有持续高表达,且与海马CA3区神经元变性坏死无相关性。“,”Objective:To detect the expression of iNOS and nNOS in delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) and their correlation with hippocampal neuron degeneration and necrosis, and to explore iNOS and nNOS in DEACMP.Methods:Seventy-two adult male SD rats were selected and randomLy(random number) divided into the DEACMP group and normal group, with 36 rats in each group. The rats were given intraperitoneal injection of 99.99% CO gas without intervention. According to different time periods before and after dying and modeling, the two groups were divided into 6 subgroups: pre-modeling, 1 d modeling, 7 d modeling, 14 d modeling, 21 d modeling, and 28 d modeling. In 6 subgroups during the modeling time, HE staining was performed to observe neuron degeneration and necrosis in hippocampal CA3 area, and immunohistochemistry and Western blot were performed to detect the protein expression of iNOS and nNOS in hippocampus. Statistical analysis was performed using SPSS 21.0 software. The measurement data were expressed as Mean±SD, and normality test and variance analysis were performed on the experimental results of each group. The mean comparison between each group adopted the Student’s t test of two independent samples. Correlation analysis was conducted between the relative expression of iNOS and nNOS protein and the degenerative necrotic neurons, Pearson correlation analysis was used for normal distribution, and Spearman rank correlation analysis was used for non-normal distribution. A n P0.05), while there were significant differences between the two groups on 14 d, 21 d, and 28 d modeling (n P0.05), while there were statistical differences between the 1 d modeling, 7 d modeling, and 14 d modeling (n P0.05), while there were statistically significant differences between the 7 d modeling, 14 d modeling, 21 d modeling and 28 d modeling (n P<0.05). Correlation analysis between the expression of iNOS protein and the count of degenerated and necrotic neurons showed a positive correlation (n P0.05).n Conclusions:iNOS plays an important role in the pathogenesis of DEACMP; nNOS is not consistently highly expressed in the hippocampus of DEACMP, and has no correlation with neuronal degeneration and necrosis in the CA3 region of the hippocampus.
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