补肾止颤方抑制帕金森病大鼠神经元细胞焦亡的作用机制

Mechanism of Bushen Zhichan Formula inhibiting pyrophosis of neurons inratswithParkinson'sdisease

目的:探讨补肾止颤方对帕金森病(PD)大鼠半胱天冬蛋白酶-1(Caspase-1)通路的作用机制。方法:将60只SD大鼠随机分为对照组,模型组,美多芭组,中药低、中、高剂量组,每组10只。采用蛋白酶抑制因子(PSI)制备PD模型,造模成功后,连续灌胃21 d治疗,期间进行行为学测试,于治疗结束后取大鼠中脑黑质区观察神经元;Western blot法检测α-突触核蛋白(α-syn)、Caspase-1、GSDMD、白细胞介素-1β(IL-1β)的表达;流式细胞术检测细胞焦亡率。结果:经中药治疗后,PD大鼠运动能力明显改善;HE染色镜下见黑质区神经元数量、形态改善;α-syn、Caspase-1、GSDMD、IL-1β蛋白表达均显著降低(P<0.05),细胞焦亡率降低(P<0.05)。结论:补肾止颤方呈剂量依赖地改善PD大鼠行为学表现及神经元受损情况,其神经保护机制与抑制Caspase-1、下调GSDMD蛋白表达、减少IL-1β的生成,从而减轻细胞焦亡相关。

Objective:To investigate the mechanism of Bushen Zhichan Decoction on Caspase-1 pathway in rats with Parkinson's disease(PD).Methods:Sixty SD rats were randomly divided into 6 groups,including control group,model group,medoba group,Chinese medicine low,medium and high dose group,with 10 rats in each group.Proteasome inhibitors I(PSI)were used to prepare PD models.After modeling,rats were given continuous intragastric treatment for 21 days,during which behavioral tests were performed,and neurons in the substantia nigra region of the middle brain were observed after treatment.The expressions of α-syn,Caspase-1,GSDMD and IL-1β were detected by Western blot.The pyroptosis rate was measured by flow cytometry.Results:The exercise ability of PD rats was significantly improved after TCM treatment.HE staining showed that the number and morphology of neurons in substantia nigra were improved.The expressions ofα-syn,Caspase-1,GSDMD and IL-1β were significantly decreased(P<0.05),and the pyroptosis rate was significantly decreased(P<0.05).Conclusion:Bushen Zhichan Decoction can improve the behavioral performance and neuron damage in a dose-dependent manner in PD rats.The neuroprotective mechanism is associated with inhibition of Caspase-1,down-regulation of GSDMD protein expression,and reduction of IL-1β production,thereby reducing pyroptosis.