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目的:研究石菖蒲挥发油有效成分β-细辛醚联合左旋多巴对6-羟基多巴诱导帕金森病模型大鼠自噬相关因子的影响。方法:将SPF级SD大鼠在内侧前脑束部位注入6-OHDA制备PD模型。将成功的PD模型大鼠随机分为模型组、雷帕霉素组(1 mg/kg)、3-甲基腺嘌呤(3-Methyl adenine,3MA)组(500 nmol/只)、美多巴组(75 mg/kg)、左旋多巴组(60 mg/kg)、β-细辛醚组(15 mg/kg)和联合用药组(左旋多巴60 mg/kg+β-细辛醚15 mg/kg),给药30 d,每天2次。给药末次1 h后,麻醉并灌注各组大鼠,取中脑做免疫荧光和流式检测Beclin-1、LC3B和p62的表达。结果:在免疫荧光和流式细胞术的检测方法中,与假手术组比较,模型组大鼠的Beclin-1和LC3B蛋白表达显著增加(P<0.01),而p62蛋白表达显著减少(P<0.01);与模型组比较,美多巴组、左旋多巴组、β-细辛醚组和联合用药组大鼠的Beclin-1和LC3B蛋白表达显著减少(P<0.05),而p62蛋白表达显著增加(P<0.05),其中β-细辛醚组和联合用药组大鼠的Beclin-1和LC3B蛋白表达显著低于美多巴组和左旋多巴组(P<0.05),而β-细辛醚组和联合用药组大鼠的p62蛋白表达显著高于美多巴组和左旋多巴组(P<0.05)。结论:β-细辛醚与左旋多巴联合应用能减少6-羟基多巴诱导PD大鼠模型中脑的自噬因子Beclin-1和LC3B表达的作用,同时增加p62的表达。
OBJECTIVE: To study the effect of β-asarone combined with levodopa, an active ingredient in Acorus calamus volatile oil, on the related factors of autophagy in rats with Parkinson’s disease induced by 6-hydroxydopamine. Methods: SP model SD rats were injected with 6-OHDA into medial forebrain to prepare PD model. The successful PD model rats were randomly divided into model group, rapamycin group (1 mg / kg), 3-methyladenine (3MA) group (500 nmol / (75 mg / kg), levodopa (60 mg / kg), β-asarone (15 mg / kg) and combination group (levodopa 60 mg / kg + mg / kg) for 30 days twice daily. After the last 1 h of administration, the rats in each group were anesthetized and perfused, and the midbrain was used for immunofluorescence and flow cytometry to detect the expression of Beclin-1, LC3B and p62. Results: Compared with the sham operation group, the expression of Beclin-1 and LC3B in the model group increased significantly (P <0.01) and the expression of p62 protein decreased significantly in the immunofluorescence and flow cytometry (P < 0.01). Compared with the model group, the expression of Beclin-1 and LC3B protein in the meduoba group, levodopa group, β-asarone group and combination group were significantly decreased (P <0.05), while p62 protein expression (P <0.05). The expressions of Beclin-1 and LC3B proteins in β-asarone group and combination group were significantly lower than those in the merida group and levodopa group (P <0.05) The p62 protein expression of asarone group and combination group was significantly higher than that of the meridional group and the levodopa group (P <0.05). CONCLUSION: The combination of β-asarone and levodopa can reduce the expression of autophagy factors Beclin-1 and LC3B in brain of PD rats induced by 6-hydroxydopamine and increase the expression of p62.