癫痫持续状态模型中海马CA1区CB2R与自噬相关蛋白LC3表达的相关性分析

The correlation between CB2R and autophagy-related protein LC3 expression in the hippocampal CA1 region of the model with status epilepticus

目的观察大鼠癫痫持续状态(status epilepticus,SE)模型中海马CA1区2型大麻素受体(cannabinoid receptor type 2,CB2R)和自噬相关蛋白微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)的表达,并探讨两者的相关性.方法使用SD大鼠腹腔注射氯化锂-匹鲁卡品建立SE模型,并随机分为4组(即对照组及造模后3 h、24 h、3 d组),应用双重免疫荧光和蛋白印迹法观察SE后不同时间点大鼠海马CA1区CB2R和LC3的表达,后使用Spearman相关性分析比较两者的相关性.结果免疫荧光结果发现CB2R在SE后呈现动态高表达,并于SE后24 h达到高峰,整体随时间呈现抛物线状变化,而LC3与CB2R变化趋势基本相同,且存在共定位表达,主要表达在神经元中;蛋白印迹实验结果进一步验证了CB2R和LC3的相似表达趋势,Spearman相关性分析发现两者表达趋势具有显著相关性(r=0.7161,P<0.05).结论SE后大鼠海马CA1区神经元CB2R和LC3的表达具有显著相关性,CB2R可能参与调节SE后海马神经元的自噬过程.

Objective The expression and correlation between cannabinoid receptor type 2(CB2R)and autophagy-related protein microtubule-associated protein 1 light chain 3(LC3)in the hippocampal CA1 region of developing rats with status epilepticus(SE)were investigated.Methods SE model was established using lithium chloride-pilocarpine intraperitoneally in Sprague-Dawley(SD)rats and all the rats were randomly divided into four groups(control group and 3h,24h,3d groups after SE).The expressions of CB2R and LC3 in the hippocampal CA1 region at different times were observed using double-label immunofluorescence and Western blotting.Spearman correlational analysis was used to compare the relationship between the two factors.Results Immunofluorescence showed that the expression of CB2R was up-regulated dynamically and peaked at 24h and presented parabolic changes over time.The expression of LC3 changed in accordance with CB2R and even co-expressed with CB2R partly,especially on neurons.Western blotting results furtherly showed the similarity of the expression of CB2R and LC3,and finally Spearman correlational analysis presented the significant correlation between these two factors(r=0.7161,P<0.05).Conclusion Significant correlation exists between the expression of CB2R and LC3 in the hippocampal CA1 neurons of developing rats with SE,indicating the essential role of CB2R in autophagy regulation of hippocampal CA1 neurons.