基于益肾填精法的茸菖胶囊对无镁诱导癫痫模型自噬相关蛋白影响的研究

[目的]探讨中药茸菖胶囊抗癫痫和神经保护作用的自噬调控机制,为临床应用提供实验依据。[方法]将无镁诱导的新生SD大鼠海马神经元放电模型分为模型组、3-甲基腺嘌呤(3-MA)组、5%茸菖胶囊组、10%茸菖胶囊组、20%茸菖胶囊组,另设正常组,给予相应处理6、12、24、72 h后,采用蛋白免疫印迹(Western blot)法检测自噬活性相关蛋白Beclin-1、B细胞淋巴瘤-2(Bcl-2)的表达。[结果]1)Beclin-1蛋白:4个时间点模型组较正常组无显著变化(P0.05),6 h 20%茸菖胶囊组及72 h 5%茸菖胶囊组较同时点模型组显著升高(P0.05)。2)Bcl-2蛋白:6 h模型组较正常组显著降低(P0.05);12、24 h 20%茸菖胶囊组,72 h 5%、10%及20%茸菖胶囊组较模型组显著增强(P0.05或P0.01),20%茸菖胶囊组较5%及10%茸菖胶囊组增强(P0.05)。[结论]基于益肾填精法的茸菖胶囊对无镁诱导的癫痫模型神经元抗癫痫及神经元保护作用的机制可能与影响Beclin-1蛋白的表达干预自噬过程,促进Bcl-2蛋白的表达抑制细胞自噬有关。

Effects of Rongchang capsule on autophagy-related proteins in epilepsy model induced by magnesium-free-treatment based on enriching kidney essence method

[Objective] To explore the autophagy regulation mechanism of antiepileptic and neuroprotective effects of Rongchang capsule to provide experimental evidence for clinical application.[Methods] Primary cultured neurons from hippocampus of neonate rats with recurrent epileptiform discharges induced by magnesium-free-treatment were divided into model group, 3-Methyladenine (3-MA) group, 5% concentration Rongchang capsule group, 10% concentration of Rongchang capsule group, 20% concentration of Rongchang capsule group, in addition, a normal group was set up. After giving corresponding treatment 6, 12, 24, 72 h, Western blot method was used to detect changes of the autophagy associated proteins (Beclin-1, Bcl-2, LC3B) expression.[Results] 1) Beclin-1 protein:6, 12, 24, 72 h Beclin-1 protein expression had no difference between model group and normal treatment group(P>0.05),6 h 20% concentration of the Rongchang capsule group and 72 h 5% concentration of the Rongchang capsule group enhanced significantly compared with model group(P<0.05). 2) Bcl-2 protein:6 h the model group Bcl-2 protein expression decreased significantly compared with the nomal group(P<0.05). 12, 24 h 20% concentration of Rongchang capsule group, and 72 h 5%, 10% and 20% concentration of Rongchang capsule group Bcl-2 protein expression enhanced significantly compared with model group (P<0.05 or P<0.01),20% concentration of the Rongchang capsule group Bcl-2 protein expression enhanced compared with 5% and 10% concentration of the Rongchang capsule group (P<0.05).[Conclusion] Rongchang capsule based on enriching kidney essence method may affect Beclin-1 protein expression and enhance Bc-l2 protein expression to play a role of antiepileptic and neuroprotective effects.