摘要
背景:研究发现星形胶质细胞中Ca^(2+)的表达与认知功能密切相关,目前由1,4,5-三磷酸肌醇受体(Inositol 1,4,5-trisphosphate receptors,IP3Rs)2、兰尼碱受体(ryanodine receptors,RYRs)2受体及其调控的Ca^(2+)信号通路已经成为认知障碍相关疾病研究的热点。目的:研究急性一氧化碳中毒迟发性脑病动物模型中,海马组织内星形胶质细胞和IP3R2、RYR2介导的Ca^(2+)信号的表达情况,探索急性一氧化碳中毒迟发性脑病可能的发病机制。方法:Morris水迷宫实验筛选认知功能合格的C57BL小鼠随机分为对照组、实验组,实验组小鼠采用静态吸入一氧化碳建立急性一氧化碳中毒迟发性脑病模型,对照组小鼠吸入等量空气。造模后第21天,利用Morris水迷宫、苏木精-伊红染色、Western blot、免疫荧光双标法、Ca^(2+)荧光探针检测中毒小鼠行为学及神经元改变、星形胶质细胞特异性标记物胶质纤维酸性蛋白及IP3R2、RYR2受体、星形胶质细胞内Ca^(2+)浓度变化。结果与结论:①Morris水迷宫实验发现与对照组相比,实验组小鼠逃避潜伏期明显延长(P<0.05);②苏木精-伊红染色表明实验组小鼠海马锥体细胞数减少、细胞结构紊乱、细胞核碎裂伴溶解;③免疫荧光检测表明海马区IP3R2、RYR2分别和胶质纤维酸性蛋白存在共表达,且实验组海马区IP3R2、RYR2和胶质纤维酸性蛋白表达均上调(P<0.05);④Western blot检测显示实验组海马区IP3R2、RYR2及胶质纤维酸性蛋白的蛋白表达均增多(P<0.05);⑤Ca^(2+)荧光探针法检测表明实验组小鼠海马区星形胶质细胞内Ca^(2+)浓度明显升高(P<0.05);⑥结果说明,星形胶质细胞可能通过介导IP3R2、RYR2受体影响Ca^(2+)信号,进而使一氧化碳中毒的小鼠认知功能受损,最终导致急性一氧化碳中毒迟发性脑病发生。
BACKGROUND:Ca^(2+)expression in astrocytes has been found to be closely related to cognitive function,and the Ca^(2+)signaling pathway regulated by inositol 1,4,5-trisphosphate receptors(IP3R2)and ryanodine receptor(RYR)2 receptors has become a hot spot in the study of cognitive disorder-related diseases.OBJECTIVE:To investigate the expression of Ca^(2+)signals mediated by IP3R2 and RYR2 in hippocampal astrocytes in animal models of delayed encephalopathy after acute carbon monoxide poisoning,and to explore the possible pathogenesis of delayed encephalopathy after acute carbon monoxide poisoning.METHODS:C57BL mice with qualified cognitive function were selected by Morris water maze experiment and randomly divided into control group and experimental group.An animal model of delayed encephalopathy after acute carbon monoxide poisoning was established by static carbon monoxide inhalation in the experimental group,and the same amount of air was inhaled in the control group.Behavioral and neuronal changes,astrocyte specific marker glial fibrillary acidic protein,IP3R2,RYR2 receptor and Ca^(2+)concentration in astrocytes of the two groups were detected using Morris water maze,hematoxylin-eosin staining,western blot,immunofluorescence double labeling and Ca^(2+)fluorescence probe at 21 days after modeling.RESULTS AND CONCLUSION:In the Morris water maze,the escape latency of the experimental group was significantly longer than that of the control group(P<0.05).Hematoxylin-eosin staining results showed that in the experimental group,the number of hippocampal pyramidal cells decreased,the cell structure was disordered,and the nucleus was broken and dissolved.Immunofluorescence results showed that IP3R2 and RYR2 were co-expressed with glial fibrillary acidic protein in the hippocampus,and the expressions of IP3R2,RYR2 and glial fibrillary acidic protein were up-regulated in the hippocampus of the experimental group(P<0.05).Western blot analysis showed that the expressions of IP3R2,RYR2,and glial fibrillary acidic protein in the hippocampus of the experimental group were increased(P<0.05).Ca^(2+)concentration in hippocampal astrocytes increased significantly in the experimental group(P<0.05).To conclude,astrocytes may affect Ca^(2+)signals by mediating IP3R2 and RYR2 receptors,then impair the cognitive function of mice with carbon monoxide poisoning,and eventually lead to delayed encephalopathy after acute carbon monoxide poisoning.
作者
赵吉利
孟天予
岳雅蓉
张鑫
杜文倩
张鑫宇
薛慧
项文平
Zhao Jili;Meng Tianyu;Yue Yarong;Zhang Xin;Du Wenqian;Zhang Xinyu;Xue Hui;Xiang Wenping(Clinical School of Medicine,Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014040,Inner Mongolia Autonomous Region,China;Department of Neurology,Baotou Central Hospital,Baotou 014040,Inner Mongolia Autonomous Region,China;Baotou Mongolian Medicine Hospital,Baotou 014040,Inner Mongolia Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2025年第2期254-261,共8页
Chinese Journal of Tissue Engineering Research
基金
中央引导地方科技发展资金(2021ZY0038),项目负责人:项文平。
