益肾达络饮治疗多发性硬化的神经免疫机制研究

【作者】 尚晓玲；

【导师】 高颖；

【作者基本信息】 北京中医药大学 ， 中医内科学， 2006， 博士

【摘要】 多发性硬化(Multiple Sclerosis,MS)是一种主要累及中枢神经系统(CNS)白质并导致多部位髓鞘脱失的自身免疫性疾病,具有易复发和致残率高的特点,属于神经系统疑难病。可在神经系统中留有病灶,形成不可逆性神经功能障碍,严重影响患者生存质量。本病在北欧及北美较常见,在非洲和东方人中发病率较低。随着诊断手段的不断改进,近年来MS的发病率在我国有明显增高的趋势。本文以中医和西医对MS的发病机制及治疗概况为背景,通过对与该病有关的文献资料的复习,从中医理论角度探讨多发性硬化的发病机理。从本病发病学、病变定位的不同方面,阐发了肾精不足,脏腑功能失调,复受外邪,络损髓伤的基本病机特点及“益肾,解毒,通络”的治法思路;以著名古方地黄饮子的配伍和功效特点为依据,在此方的基础上创立益肾达络饮作为本病的治疗方剂,并进行初步实验研究。本研究包括理论探讨和实验研究二部分:多发性硬化的病因病机理论探讨;多发性硬化的实验研究。研究一多发性硬化的病因病机理论探讨围绕多发性硬化的发病进程,从病因出发,结合疾病不同发病时期的特点深入探讨病机,阐述不同时期的病机变化。发作期内外因相合,肾精不足、毒邪外侵综合作用于人体,导致督脉、脑、肾同病。缓解期以正虚为主,督脉不充,肾阳不足,脑髓失养。复发期再感邪气或外邪引动旧邪复燃,毒侵督脉,损伤脑髓。以脏腑与经络相结合,提出本病是在肾精不足,脏腑功能失调基础上,湿浊内蕴,浊毒内生,毒损督脉,戕害脑髓,络损髓伤,败坏形体而发病的病机理论。研究二多发性硬化的实验研究本实验研究旨在通过制备MS动物模型,在中药干预的前提下,研究其临床症状、体征评价、病理改变观察,为中医药治疗MS提供现代生物学依据。课题通过经典动物模型实验性变态反应性脑脊髓炎(EAE)小鼠,采用HE染色、免疫组化、ELISA、原位杂交等实验方法,配合光镜、图像分析等细胞定性、半定量检测手段,从神经生物学分子水平探讨EAE病变过程中关键环节细胞因子的变化,并在此基础上观察益肾达络饮对EAE的影响及对免疫功能的调节作用。以期在从基因分子水平探讨EAE发病机制的同时,从治疗和康复角度研究益肾达络饮对EAE的作用机理。本研究选择SJL小鼠用PLP139-151诱导制备EAE复发-缓解模型。每天观察小鼠神经功能评分,并分别用醋酸泼尼松、益肾达络饮给予灌胃治疗14天,在造模后22天,即症状高峰期取材。通过ELISA、免疫组化、原位杂交等实验检测方法,探讨EAE病理机制。从中枢神经组织内T细胞亚群变化、趋化因子、黏附分子、轴索病变、p38等几方面着重观察了EAE病理改变及中药复方益肾达络饮对EAE的影响,探讨EAE发病机制,从轴索和髓鞘同时明确益肾达络饮对改善神经功能的作用机制。并进一步从神经免疫角度来揭示益肾达络饮对EAE神经细胞及轴索的保护作用,及其潜在的治疗MS的更多还原

【Abstract】 Multiple Sclerosis(MS) is chronic inflammatory and demyelinating disorder disease involving white matter with the numerous affected areas of central nervous system possibly due to autoimmunity. The patients subjected to multiple sclerosis typically show alternating relapse and remission in the early stage of illness. MS is the one of the leading causes of neurological disability in young adults. It is regarded as stubborn disease. Producing the characteristic plaques or sclerosed areas that are the hallmark of the disease, MS has the serious effect to patient’s living quality because of irreversibility nervous functional impairment. The disease incidence of MS is much more higher in Northern Europe and North America than Africa and oriental area. With the unceasing improvement of diagnosis instrument, the disease incidence of MS recently shows a ascending tendency obviously in China.On the basis of the overview to pathogenesy and treatment of Traditional Chinese Medicine(TCM) and Western medicine for MS, MS pathogenesy is approached at the point of view on TCM. From the aspects of pathogenesis and pathological changes localization, we interpret the basic pathogenesis characteristic that is brain marrow and collateral injury due to deficiency of kidney-essence, function disharmony of entrails, recurrence of exogenous evil, and the direction to treatment according to tonifying kidney, removing toxic substance, removing obstruction in collaterals.On the basis of the feature of compatibility and efficacy of the distinguished ancient prescription-dihuang yin, we bring forward Yi Shen Da Luo Decoction as the treantment prescription to MS, and carry out initial empirical study of its animal models- experimental allergic encephalomyelitis (EAE).This research was consisted of two parts: Discussion on the theory of pathogeny and pathogenesis of Multiple Sclerosis;The empirical study on EAE mouse.Part one Discussion on the theory of pathogeny and pathogenesis of MSFrom the view of TCM, the pathologic mechanism of MS was discussed encircling pathogeny and the course, in order to explain the pathologic mechanism change combined with characteristic in different phase of MS. With the connection of zang-fu viscera and channel, we put forward the pathogenesis theory. On the basis of insufficiency of kidney essence and function disharmony of entrails, turbid dampness and toxin accumulated, Governor Channel was impaired by pathogenic factors and brain marrow was slayed with injury in collaterals and marrow.更多还原