从miRNA调控探讨草果知母汤治疗癫痫的作用机制

摘要

Objective: To explore the mechanism of Caoguo Zhimu decoction in treating epilepsy. Methods: 60 SPF SD male rats were randomly divided into blank control group, model group, Caoguo zhimu decoction low, medium and high dose groups with12 rats in each group. Except for control group, the other 4 groups all prepared epilepsy models and administered them after 1 hour.Control group and model group were given 2 m L/kg of double distilled water while Caoguo zhimu decoction low, medium and high dose groups were administered with herbal extracts of 0.375, 0.75, 1.5 g/m L, 2 m L/kg, respectively, once a day for 28 days. The behavioral changes of epileptic rats in each group were observed. Histopathological observation of hippocampal tissue by HE staining.TUNEL method was used to observe neuronal apoptosis. RT-PCR was used to detect the expression of mi R-21, mi R-29 and apoptosis related genes Bax, Bcl-2 and p53 m RNA. Western Blot was used to detect the expression of Bax, Bcl-2 and p53 protein. Results:Caoguo Zhimu decoction was able to improve epilepsy behavior of epileptic rats in all dose groups and positive drug groups, reduce neuronal apoptosis in hippocampal tissue, reduce the expression of mi R-29, Bax and P53 m RNA (P＜0.05 or 0.01) and increase the expression of mi R-21 and Bcl-2 RNA protein (P＜0.05 or 0.01), reduce the expression of Bax, P53 protein (P＜0.01), increase the expression of Bcl-2 protein (P＜0.01). Conclusion: Regulating mi R-21, mi R-29 to adjust the apoptosis related genes Bax, Bcl-2 and p53 expression might be one of the mechanism of Caoguo Zhimu decoction in treating epilepsy.%目的 探讨草果知母汤治疗癫痫的作用机制.方法 选取SPF级SD雄性大鼠60只,随机分为正常组,模型组,草果知母汤低、中、高剂量组,每组12只.除正常组外,其余4组均制备癫痫模型,造模1 h后给药,正常组、模型组以双蒸水2 m L/kg灌胃,草果知母汤低、中、高剂量组分别以草果知母汤0.375、0.75、1.5 g/m L生药提取液2 m L/kg灌胃,每日1次,持续28 d.观察各组大鼠癫痫行为学变化,取各组大鼠脑海马组织HE染色进行组织病理学观察,以TUNEL法观察神经细胞凋亡情况,RT-PCR检测mi R-21、mi R-29和凋亡相关基因Bax、Bcl-2、P53 m RNA表达,Wester Blot法检测凋亡相关基因Bax、Bcl-2、P53蛋白表达.结果 草果知母汤各剂量组均有改善癫痫大鼠癫痫行为学的作用,可减少海马组织神经细胞凋亡,减少mi R-29、Bax和P53 m RNA的表达(P＜0.05或0.01),增加mi R-21和Bcl-2m RNA的表达(P＜0.05或0.01),降低Bax、P53蛋白表达(P＜0.01)并增加Bcl-2蛋白表达(P＜0.01).结论 通过调控mi R-21、mi R-29进而调节凋亡相关基因Bax、Bcl-2、P53的表达,可能是草果知母汤治疗癫痫的作用机制之一.