摘要： 目的 探讨瞬时受体电位通道7 （TRPM7）在癫痫中与凋亡的关系。方法 采用CCK-8法检测匹鲁卡品（PILO）对Neuro-2A细胞活性的影响；设置对照组和PILO模型组，采用蛋白质印迹法检测凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶3 （caspase-3）、Bcl-2相关X蛋白（Bax）和细胞色素C蛋白（Cyt C）的表达；采用实时荧光定量PCR法检测TRPM7 mRNA的表达；设置对照组（Neuro-2A细胞转染无序siRNA序列）、PILO模型组（Neuro-2A细胞转染无序siRNA序列后经PILO处理24 h）和PILO+siRNA组（Neuro-2A细胞转染TRPM7 siRNA序列后经PILO处理24 h），采用蛋白质印迹法检测凋亡相关蛋白表达；采用TUNEL染色观察细胞凋亡情况。结果 Neuro-2A细胞活性随PILO浓度升高而降低；与正常对照组相比，模型组凋亡相关蛋白caspase-3、Bax、Cyt C和TRPM7 mRNA在24 h时表达明显增加（均P < 0.05）；与PILO模型组相比，PILO+siRNA组凋亡相关蛋白caspase-3、Bax和Cyt C表达和细胞凋亡率明显减少（均P < 0.05）。结论 TRPM7参与PILO诱导的癫痫体外模型的凋亡。

瞬时受体电位通道7参与匹鲁卡品诱导的癫痫体外模型凋亡的调控

李晓娜¹, 何苗², 梁洪玥², 史瑞雪², 付钰², 顾宇婧², 李艾凝², 郭凤²

1. 中国医科大学附属第四医院药学部, 沈阳 110032;
2. 中国医科大学药学院药物毒理学教研室, 沈阳 110122

收稿日期:2021-09-06出版日期:2022-04-20发布日期:2022-05-05
通讯作者:郭凤E-mail:aforge@126.com
作者简介:李晓娜(1985-),女,药师,本科.
基金资助:国家自然科学基金 （81971212）


关键词: 癫痫, 瞬时受体电位通道, 凋亡, 匹鲁卡品, 小鼠脑神经瘤细胞
Abstract: Objective To investigate the relationship between transient receptor potential melastatin 7 (TRPM7) and apoptosis in epilepsy. Methods Neuro-2A cell viability was assessed by the cell counting kit-8 (CCK-8) assay. The expressions of caspase-3 protein (caspase-3),Bcl-2 related X protein (Bax),and cytochrome C protein (Cyt C) were determined by Western blotting in the control group and model group. The mRNA expression level of TRPM7 was detected by real-time PCR. Additionally,the protein expressions of caspase-3,Bax,and Cyt C were determined by Western blotting in the control group,pilocarpine (PILO) model group,and PILO+siRNA group. The apoptotic rate was detected by TUNEL staining in the control group,PILO model group,and PILO+TRPM7 siRNA group. Results The results showed that the viability of Neuro-2A cells decreased with increasing PILO concentrations. Moreover,the protein expressions of caspase-3,Bax,and Cyt C and mRNA expression of TRPM7 increased in the PILO group compared with the control group (all P < 0.05). Meanwhile,the protein expressions of caspase-3,Bax,and Cyt C and the apoptotic rate decreased in the PILO+siRNA group when compared with those of the PILO model group (all P < 0.05).Conclusion TRPM7 was involved in the modulation of apoptosis in epilepsy in an in vitro model induced by PILO.
Key words: epilepsy, transient receptor potential channel, apoptosis, pilocarpine, neuro-2A cell
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