从泛素-蛋白酶体系统及内质网应激探讨祛风湿类中药对帕金森病多巴胺能神经元的保护机制

Parkinson's disease(PD) is the second largest neurodegenerative disease in the middle-aged and its pathological characteristics are loss of dopaminergic neurons in the substantia nigra and whith formation of Lewy body (LB) .LB is mainly within a-Synuclein, ubiquitin protein. Recent research has shown that LB formation is closely related to the functional defect of ubiquitin proteasome system (UPS) , UPS is an important pathway of abnormal protein degradation in vivo;And regulations of protein processing, folding and degradation are closely related to Endoplasmic reticulum stress(ERS);Inflammatory cytokines can activate the ERS;UPS and ERS dysfunction can lead to the accumulation of abnormal proteins in cells and even cell death(apoptosis).Duhuo is a kind of wind -damp dispelling herbs .Throughtout the records of generations of doctors,it can tonify the five zang-organs, especially the kidney, dispelling static blood and phlegm, and dredging cerebral vascellum.Early research has shown that Duhuo coumarin could improve abnormal behavior of Lactacystin-induced PD model rats by regulating immune function and resisting ioxidative damage ,excitatory aminoacid glutamate toxicity and apoptosis of dopaminergic neurons in the substantia nigra.This research, through experiments in vivo and in vitro ,aims to observe the influence of the wind-damp dispelling herbs on ESR, UPS, inflammatory factors and apoptosis factors ,and to explore its possible mechanisms of protecting DA neurons.?????.

帕金森病（PD）是中老年第二大神经变性疾病，其病理特征是黑质多巴胺能神经元变性缺失伴Lewy小体（LB）的形成。LB内主要有a-突触核蛋白 、泛素等。近来研究表明：LB形成与泛素-蛋白酶体系统(UPS)功能缺损密切相关，UPS是体内异常蛋白降解的重要通路；而蛋白加工、折叠、降解的调控又与内质网应激(ERS)有密切关系；炎症因子可激活ERS；UPS和ERS功能异常可导致细胞内异常蛋白的蓄积，甚至细胞死亡（凋亡）。中药独活为祛风湿类药，纵观历代医家记载，独活还具有调补五脏，尤善补肾，祛瘀化痰，宣通脑络功效。前期研究表明，独活香豆素能通过调节免疫功能、抗氧化损伤和兴奋性氨基酸谷氨酸毒性及降低黑质多巴胺能神经元的凋亡而改善Lactacystin诱导的PD模型大鼠行为学异常。本研究拟通过体内、外实验，观察祛风湿类中药对ESR、UPS及炎症因子、凋亡因子影响，探讨其保护多巴胺能神经元的可能机制。

帕金森病（PD）是常见于中老年的神经系统变性疾病，发病率高，其静止性震颤、肌强直等行为异常严重影响患者的生活质量,黑质多巴胺能神经元变性丢失和路易小体的形成是PD主要病理特征，a-Syn是路易小体的主要成分。近来研究发现：PD的发生与泛素-蛋白酶体系统(UPS)功能缺损、内质网应激(ERS)、炎症因子异常及细胞凋亡关系密切。中药独活为祛风湿类中药之首，纵观历代医家记载，独活还具有调补五脏，尤善补肾，祛瘀化痰，宣通脑络功效。临床应用其治疗PD效果较好。为探讨其作用机制，本研究通过体内、外实验，应用免疫荧光(激光共聚焦)、RT-PCR、Western、Elisa方法，观察其对UPS、ESR、炎症因子及凋亡因子的影响。结果发现：其能改善PD模型大鼠的行为异常，提高脑黒质的TH细胞数量；提高Lactacystin诱导的PC12的细胞存活率；上调PD模型（大鼠脑黒质和/PC12）UPS酶类UBEI、UCHL-1mRNA表达，减弱a-Syn阳性细胞表达；下调ESR相关因子 CHOP、Caspase12蛋白和mRNA表达；下调炎症因子IL-1α、IL-1β、IL-6 、TNF-α蛋白和mRNA表达；下调凋亡因子Fas、Fas-L、Cyt-c、caspase-3蛋白和mRNA表达。提示独活改善PD模型大鼠行为学异常、减少神经元损伤，与其改善UPS功能缺损、从而减轻ERS、抑制炎症因子异常和细胞凋亡密切相关。目前PD治疗多采用左旋多巴为主的替代疗法，其虽能改善症状,但不能延缓和逆转病程，且长期服用会产生疗效减退及难以纠正的症状波动和异动症等副作用。祛风湿类中药独活无明显毒付作用，治疗PD符合中医病因病机和治病求本理论；并显示出上述机制的调控作用；且现代研究发现它还具有延缓衰老、抗肿瘤、抗心血管疾病等多种作用，因此其研究具有科学意义及广阔的应用前景。