作者：傅永旺，张玉，武艳芳

单位：

010017内蒙古自治区呼和浩特市，内蒙古自治区人民医院医学高压氧治疗科

Units：

Department of Hyperbaric Oxygen, Inner Mongolia People's Hospital, Hohhot 010017, China

关键词：

一氧化碳中毒迟发性脑病；大鼠；海马；细胞凋亡；线粒体

Keywords：

Delayed carbon monoxide encephalopathy; Rats; Hippocampus; Apoptosis; Mitochondria

CLC：

　R 595

DOI：

10.12114/j.issn.1008-5971.2022.00.268

Funds：

内蒙古自治区自然科学基金资助项目（2021LHMS08063）；内蒙古自治区卫生健康科技计划项目（202201051）；内蒙古自治区人民医院基金项目（2019YN07）

摘要：

目的　探究一氧化碳中毒迟发性脑病（DEACMP）大鼠海马组织神经元凋亡情况及机制。方法　本实验时间为2021年3月，选取6周龄雄性SPF级SD大鼠30只，随机分为对照组（n=6）和模型组（n=24）。模型组大鼠采用自然吸入一氧化碳中毒方法制备DEACMP模型，对照组大鼠仅进行空气暴露。采用Morris水迷宫实验判断大鼠是否发生DEACMP，模型组16只存活的大鼠中发生DEACMP 7只（DEACMP组），未发生DEACMP 9只（无DEACMP组）。从对照组、无DEACMP组、DEACMP组中各选取6只大鼠，采用TUNEL法检测大鼠海马组织神经元凋亡情况，采用超氧化物阴离子荧光染色法检测大鼠海马组织活性氧表达情况，采用Western blot法检测大鼠海马组织线粒体相关凋亡因子〔半胱氨酸天冬氨酸蛋白酶（Caspase）-3、细胞色素C（CytC）、B细胞淋巴瘤蛋白2（Bcl-2）〕表达水平。结果　DEACMP组大鼠海马组织神经元凋亡指数高于无DEACMP组和对照组（P ＜0.05）；超氧化物阴离子荧光染色法结果显示，对照组、无DEACMP组、DEACMP组大鼠海马组织均未检测到活性氧。DEACMP组和无DEACMP组大鼠海马组织Caspase-3、CytC表达水平高于对照组，DEACMP组大鼠海马组织Caspase-3、CytC表达水平高于无DEACMP组（P ＜0.05）；DEACMP组和无DEACMP组大鼠海马组织Bcl-2表达水平低于对照组（P ＜0.05）。结论　DEACMP大鼠存在海马组织神经元凋亡，其机制可能与Caspase-3、CytC表达水平升高及Bcl-2表达水平降低有关。

Abstract：

　Objective　To investigate the apoptosis of hippocampal neurons in rats with delayed carbon monoxideencephalopathy (DEACME) and its mechanism. Methods　The experimental time was March 2021. Thirty 6-week-old male, SPFgrade SD rats were selected and randomly divided into control group (n=6) and model group (n=24) . The rats in the model group wereexposed to natural carbon monoxide to prepare DEACMP model, while the rats in the control group were exposed to air only. Morriswater maze test was used to determine whether DEACMP occurred in rats. Among the 16 surviving rats in the model group, 7 ratsoccurred DEACMP (DEACMP group) and 9 rats did not occurred DEACMP (no DEACMP group) . Six rats were selected from eachof the control group, no DEACMP group and DEACMP group. The apoptosis of hippocampal neurons of rat was detected by TUNELmethod, the expression of reactive oxygen species in hippocampal tissue was detected by superoxide anion fluorescence staining,and the expression of mitochondrial related apoptotic factors [cysteinyl aspartate-specific proteinase (Caspase)-3, cytochromeC (CytC), B-cell lymphoma protein 2 (Bcl-2) ] in hippocampal tissue was detected by Western blot. Results　The apoptotic indexof hippocampal neurons in DEACMP group was higher than that in DEACMP group and control group (P < 0.05) ; the results ofsuperoxide anion fluorescence staining showed that no reactive oxygen species was detected in the hippocampal tissue of the controlgroup, no DEACMP group and DEACMP group. The expression levels of Caspase-3 and CytC in the hippocampal tissue in the DEACMP group and the no DEACMP group were higher than those in the control group. The expression levels of Caspase-3 andCytC in the hippocampal tissue in the DEACMP group were higher than those in the no DEACMP group (P < 0.05) ; the expressionlevel of Bcl-2 in hippocampal tissue in DEACMP group and no DEACMP group was lower than that in control group (P < 0.05) .Conclusion　Apoptosis of hippocampal neurons in DEACMP rats may be related to the increase of Caspase-3, CytC expression leveland decrease of Bcl-2 expression level.

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