王春玲 罗宁 蒋媛静 蒙冰 左曜玮 李昌海 文晓东

摘要 目的：觀察敛肝熄风止颤方对帕金森病（Parkinson Disease，PD）的影响，探讨其神经保护机制。方法：将60只SD大鼠分为假手术组10只，造模组50只，造模组大鼠接受PD模型制备，将成模的40只大鼠随机分为模型组10只、低剂量组（0.36 g/kg）10只、中剂量组（0.72 g/kg）及高剂量组（1.44 g/kg）10只，持续灌胃给药30 d，比较各组大鼠阿扑吗啡诱导旋转次数、圆筒实验，纹状体超微结构以及CaMkkβ、AMPK、p-AMPK水平的变化。结果：1）敛肝熄风止颤方可明显减少大鼠转圈以及肢体碰壁的次数，随着剂量增加次数减少更明显，差异有统计学意义（P<0.05）。2）造模大鼠纹状体线粒体自噬现象减弱，敛肝熄风止颤方可明显促进脑组织线粒体自噬。3）中药干预后大鼠纹状体CaMkkβ、p-AMPK蛋白表达上调，与模型组比较，差异有统计学意义（P<0.05），其中高剂量组较低剂量组上调明显，差异有统计学意义（P<0.05）。结论：敛肝熄风止颤方对帕金森病具有神经保护作用，其机制之一可能是通过激活CaMkk/AMPK通路活性促进线粒体自噬有关。

关键词 帕金森病;敛肝熄风止颤方;机制;线粒体;自噬;神经保护;CaMkkβ/AMPK通路

Study on the Neuroprotective Mechanism of Lianggan Xifeng Zhichan Formula Based

on Mitochondrial Autophagy Mediated by CaMkkβ/AMPK Pathway

WANG Chunling1，LUO Ning2，JIANG Yuanjing2，MENG Bing2，ZUO Yaowei1，LI Changhai1，WEN Xiaodong2

（1 College of Pharmacy，Guangxi University of Traditional Chinese Medicine，Nanning 530001，China; 2 Department of

Neurology，Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine，Nanning 530011，China）

Abstract Objective：To observe the effect of Lianggan Xifeng Zhichan Formula （LGXFZC） on Parkinson′s disease （Parkinson′s disease，PD） and explore its neuroprotective mechanism.Methods：A total of 60 SD rats were divided into a sham operation group （n=10） and a model group （n=50）.The rats in the model group were made by PD model.40 rats were randomly divided into a model group （n=10），a low dose group （0.36 g/kg） （n=10），a middle dose group （0.72 g/kg） and a high dose group （1.44 g/kg） for 30 days.The rotation times of apomorphine，cylinder test，ultrastructure of striatum and the levels of CaMkk β，AMPK and p-AMPK were compared in each group.Results：1） LGXFZC could significantly reduce the frequency of rotation and limb touching wall in rats.As the dose increases，the frequency decreases more obviously.The difference was statistically significant （P<0.05）.2） the phenomenon of mitochondrial autophagy in striatum of model rats was weakened，and LGXFZC could obviously promote mitochondrial autophagy in brain tissue.3） after the intervention of Chinese medicine，the expression of CaMkk β and p-AMPK protein in the striatum of rats was significantly higher than that in the model group （P<0.05），and compared with the model group，the difference is statistically significant （P<0.05）.Among them，the high-dose group and the lower-dose group were up-regulated significantly （P<0.05）.Conclusion：LGXFZC has neuroprotective effect，and one of its mechanisms may be that it promotes mitochondrial autophagy by activating the activity of CaMkk/AMPK pathway.

Keywords Parkinson′s disease; Linggan Xifeng Zhichan Formula; Mechanism; Mitochondria; Autophagy; Neuroprotection; CaMkk β/AMPK pathway

中图分类号：R289.5;R741文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.05.015

帕金森病（Parkinson Disease，PD）是临床常见的神经退行性疾病，以黑质-纹状体多巴胺能神经元进行性丢失为主要病理学改变[1]，严重影响患者的日常工作和生命质量。因此，寻找安全有效的治疗PD手段，是神经医学及康复医学迫切需要解决的课题。线粒体自噬是清除/降解受损线粒体的过程，是有效阻断受损线粒体导致细胞死亡的关键[2-3]。另一方面，线粒体自噬过程中产生脂肪酸、氨基酸等小分子物质将重新参与机体组织细胞的物质代谢，有研究显示帕金森病的发生发展与线粒体自噬减弱关系密切，调控线粒体自噬对治疗PD多巴胺能神经元损伤有重要的意义[4-5]。

敛肝熄风止颤方首载于《伤寒论·厥阴病》，有敛肝熄风、养血濡筋的作用，临床显示将该方用于治疗PD，确具有一定的疗效[6]，但其机制仍需进一步研究。……