龟羚帕安丸对帕金森病大鼠转录因子EB介导自噬调控机制的研究

Effect of Guiling Pa′an Pill on the Regulation Mechanism of Transcription Factor EB-mediated Autophagy in Parkinson′s Disease Rats

目的:研究龟羚帕安丸治疗帕金森病(PD)对转录因子EB(TFEB)表达的影响及其机制。方法:将PD大鼠模型随机分为模型组、西药组、中西药组、中药低、中、高剂量组,每组15只,另取15只正常大鼠作为假手术组。模型组及假手术组给予等容积生理盐水,其余组给予相应药物灌胃,各组均连续给药4周。PD大鼠处死后,将中脑黑质组织剥离出来,运用蛋白质印迹法(Western Blotting)检测TFEB、微管相关膜蛋白轻链3Ⅱ(LC3Ⅱ)、泛素结合蛋白P62(P62)、酪氨酸羟化酶(TH)、α-突触核蛋白(α-syn)的表达量。结果:与假手术组比较,模型组、西药组、中西药组、中药各剂量组黑质内TFEB、LC3Ⅱ、TH蛋白表达显著降低(P<0.01),P62、α-syn表达显著增高(P<0.01);与模型组比较,西药组、中西药组、中药各剂量组黑质内TFEB、LC3Ⅱ、TH蛋白表达显著增高(P<0.01),P62、α-syn表达显著降低(P<0.01)。结论:龟羚帕安丸可通过TFEB激活自噬-溶酶体系统,增强神经细胞自噬活性,增加LC3Ⅱ、TH表达,促进α-syn、P62降解及清除,发挥神经保护作用。

Objective:To study the effect of Guiling Pa′an Pill on the expression of transcription factor EB(TFEB) in the treatment of Parkinson′s disease(PD) and its mechanism.Methods:The PD rat model was randomly divided into model group,western medicine group,traditional Chinese and western medicine group and traditional Chinese medicine low,middle and high dose groups,with 15 in each group.Another 15 normal rats were selected as sham operation group.The model group and sham operation group were given same volume of normal saline,and the other groups were given corresponding medicines by intragastric administration for 4 consecutive weeks.After the PD rats were sacrificed,the substantia nigra of the midbrain was collected.The protein expressions of TFEB,microtubule-associated protein light chain 3Ⅱ(LC3Ⅱ),P62,tyrosine hydroxylase(TH) and α-synuclein(α-syn) were detected by Western Blotting.Results:Compared with the conditions in sham operation group,the protein expressions of TFEB,LC3Ⅱ and TH in the substantia nigra of the model group,western medicine group,traditional Chinese and western medicine group and traditional Chinese medicine groups decreased significantly(P<0.01),while the protein expressions of P62 and α-syn increased significantly(P<0.01).Compared with the model group,the western medicine group,traditional Chinese and western medicine group and traditional Chinese medicine groups had up-regulated TFEB,LC3Ⅱ and TH in substantia nigra(P<0.01),while down-regulated P62 and α-syn(P<0.01).Conclusion:Guiling Pa′an Pill can activate autophagy-lysosome system through TFEB,enhance the autophagy of neural cells,increase LC3Ⅱ and TH,and promote the degradation and clearance of α-syn and P62,thereby playing a neuroprotective effect.