水木和宁方对鱼藤酮诱导的帕金森模型小鼠自噬功能的影响

Effect of Shuimu Hening Formulation on autophagy function of rotenone-induced Parkinson's model mice

关键词:  水木和宁方  帕金森病  鱼藤酮  自噬  作用机制

Key words:  Shuimu Hening Formulation  Parkinson's disease  rotenone  autophagy  mechanism of action

摘要:

[目的] 观察水木和宁方对帕金森模型小鼠行为学及中脑黑质内酪氨酸羟化酶（TH）、α-突触核蛋白（α-syn）、自噬相关蛋白的干预作用。[方法] 采用颈背部皮下注射鱼藤酮法制备帕金森小鼠模型，将造模成功的小鼠随机分为高剂量组、中剂量组、低剂量组、美多芭组和模型组，高、中、低剂量组分别给予水木和宁方颗粒，美多芭组予美多芭，模型组和正常组予生理盐水，连续28 d。从行为学对小鼠进行观察；通过免疫组化、免疫印迹、RT-PCR实验对小鼠中脑黑质组织TH、α-syn及自噬相关蛋白微管相关膜蛋白轻链3（LC3）、自噬启动蛋白（Beclin1）、自噬特异性底物蛋白（p62）进行检测。[结果] 1）模型组小鼠表现出运动迟缓、反应迟钝、拒捕反应减弱、毛发粗糙、凌乱、变黄，予水木和宁方及美多芭治疗后，上述症状均有所改善。2）水木和宁方和美多芭皆可提高PD模型小鼠中脑组织TH的表达，降低α-syn的表达，两者均可通过干预自噬相关蛋白达到治疗效果。3）中剂量组效果最佳，较其他组差异具有统计学意义。[结论] 水木和宁方可通过上调自噬水平，改善自噬障碍，促进自噬流的顺利完成，降解异常聚集的α-syn，从而保护多巴胺（DA）能神经元。

Abstract:

[Objective] To observe the intervention effects of Shuimu Hening Formulation on the behavior of Parkinson disease(PD) model mice and TH,alpha-syn and autophagy related proteins in the substantia nigra of the midbrain.[Methods] PD mouse model was prepared by subcutaneous injection of rotenone into the neck and back.The mice were randomly divided into high-dose group,medium-dose group,low-dose group,medoba group and model group. Shuimu Hening Formulation were given to the high,medium and low dose groups,medoba was given to themedoba group,and distilled water was given to the model group and normal group.The above groups were given for 28 consecutive days. The behavior of mice was observed. TH,α-syn and autophagy related proteins LC3,p62 and Beclin1 were determined by immunohistochemistry,western blot and RT-PCR.[Results] 1)After the model building,the behavior of mice was changed,which was mainly manifested as slow movement,slow reaction,weakened anti-arrest reaction,rough and disorderly hair,and yellow color. After the treatment of Shuimu Hening Formulation andmedoba,the above symptoms were improved. 2) Both Shuimu Hening Formulation and medoba can increase the expression of TH in midbrain tissue of PD model mice and reduce the expression of alpha-syn. Both of them can achieve the therapeutic effect by intervening autophagy related proteins. 3)The effect of the medium dose groupwas better than that of other groups.[Conclusion] These results suggest that Shuimu Hening Formulation can up-regulate the autophagy level,improve the autophagy obstacle,promote the smooth completion of autophagy flow,and degrade the abnormal aggregation of alpha-syn,thus protecting DA neurons.