治療帕金森氏症與脊髓性肌肉萎縮症之中草藥開發：六味地黃丸、馬錢子苷及牡丹酚

Herbal medicine development for Parkinson’s disease and spinal muscular atrophy: liuwei dihuang, loganin, and paeonol

曾于庭(Yu-Ting Tseng)

指導教授 ： 羅怡卿

高雄醫學大學/藥學院/天然藥物研究所博士班/博士(2016年)

摘要

由於人口老化，神經退化性疾病的藥物開發成為現今社會重要的醫療議題。然而，由於造成神經退化的病因及分子機轉極為複雜，使得有效的治療藥物開發至今仍困難重重。因此，神經退化性疾病新的治療方針傾向於發展能作用在多重標的並具有多功能之候選藥物。目前有許多神經退化性疾病可能的病理機轉被提出，包括發炎、氧化壓力、粒線體功能受損、滋養因子缺乏，以及基因變異等，而最近的研究指出許多中草藥具有抗發炎、抗氧化，以及神經滋養活性，能作為延緩疾病進展的輔助及替代藥物。本論文以帕金森氏症及脊髓性肌肉萎縮症疾病細胞及動物模式建立中草藥藥物開發平台，以現代醫學理論及方法評估中草藥包括六味地黃丸、馬錢子苷及牡丹酚的療效以及藥理機轉。帕金森氏症實驗模式使用內毒素誘發發炎之小神經膠細胞、帕金森毒物1-甲基-4-苯基?m啶離子(1-methyl-4-phenylpyridinium; MPP+)或六-羥基多巴胺(6-hydroxydopamine; 6-OHDA)誘發氧化性損傷之初代培養神經元細胞，及1-甲基-4-苯基1,2,3,6-四氫?m啶(1-methyl-4-phenyl-1,2,3,6-tetra hydropyridine; MPTP)誘發之帕金森氏症疾病小鼠。脊髓性肌肉萎縮症實驗模式使用運動神經元存活蛋白缺乏之運動神經元細胞、脊髓性肌肉萎縮症病人之皮膚纖維母細胞，以及7號外顯子缺失模式之脊髓性肌肉萎縮症小鼠(SMA7)。研究結果顯示，由熟地黃、山藥、山茱萸、澤瀉、茯苓及牡丹皮組成的六味地黃丸能降低內毒素誘發之小神經膠細胞發炎， MPP+誘發之氧化壓力及神經損傷，且在帕金森氏症疾病小鼠模式中能保護多巴胺神經元並改善小鼠運動功能。在脊髓性肌肉萎縮症細胞及動物模式中，六味地黃丸能增加運動神經元存活蛋白表現、改善運動神經元存活蛋白缺乏造成的運動神經元神經元損傷、改善病鼠的肌肉張力並增加其體重和存活。我們也利用所建立的藥物開發平台評估山茱萸中的環烯醚萜配糖體馬錢子苷的保護作用與機轉。研究結果發現，馬錢子苷能藉由活化昇糖素樣胜肽-1受體降低MPP+誘發之神經元損傷。而在脊髓性肌肉萎縮症細胞模式中，馬錢子苷能增加運動神經元存活蛋白表現，並藉由活化胰島素樣生長因子-1受體降低運動神經元存活蛋白缺乏造成的運動神經元損傷。在脊髓性肌肉萎縮症疾病動物模式中，馬錢子苷能促進蛋白質合成路徑，改善病鼠的肌肉張力，並增加其體重和存活。另一方面，我們也在發炎及氧化壓力細胞模式中評估一牡丹皮中酚類天然物牡丹酚的保護作用。研究結果顯示，牡丹酚能降低內毒素誘發之小神經膠細胞發炎，並減少小神經膠細胞發炎以及6-OHDA誘發之神經毒性。本論文建立了一帕金森氏症及脊髓性肌肉萎縮症中草藥藥物開發平台，並證實了六味地黃丸、馬錢子苷以及牡丹酚應用於此類神經退化性疾病輔助治療的潛力。同時，我們也提出了馬錢子苷未來應用於蛋白質合成路徑損傷相關之肌肉萎縮症的可能。希望本論文之研究成果，能提供中草藥在神經保護上的證據及臨床應用。也期許本論文所建立的帕金森氏症及脊髓性肌肉萎縮症藥物開發平台，能作為未來相關疾病治療中草藥開發之參考。

關鍵字

中草藥 ； 神經退化性疾病 ； 發炎 ； 氧化壓力 ； 帕金森氏症 ； 脊髓性肌肉萎縮症

並列摘要

Drugs development on neurodegenerative diseases become important medical issues due to the increase of aging population. However, complex pathology and molecular cascade of events that results in neurodegeneration makes it difficult to achieve efficiency on drug treatment. Therefore, novel therapeutic approaches of neurodegenerative diseases comprise drug candidates to act on multiple targets and process multi-functional properties. A variety of possible pathogenic mechanisms of neurodegenerative diseases have been proposed, including inflammation, oxidative stress, mitochondrial dysfunction, loss of trophic support, and gene mutation. Recent studies also suggest a large majority of herbal medicine process anti-inflammatory, anti-oxidative, and neurotrophic activities, can be considered as promising complementary and alternative approaches for disease progression of neurodegenerative diseases. The present thesis develops a drug screening platform based on disease cellular and animal models of Parkinson’s disease (PD) and spinal muscular atrophy (SMA), examining protective effects of herbal medicine including liuwei dihuang (LWDH), loganin, and paeonol, and evaluating their pharmacological mechanisms by modern medical theories and methods. In experimental models of PD, lipopolysaccharide (LPS)-induced inflammatory microglial cultures, 1-methyl-4-phenylpyridinium (MPP+) or 6-hydroxydopamine (6-OHDA)-induced oxidative damaged primary neuronal cultures, and 1-methyl-4-phenyl-1,2,3,6-tetra hydropyridine (MPTP)-induced PD mice were used. In experimental models of SMA, survival motor neuron protein (SMN)-deficient motor neuronal cultures, skin fibroblasts from SMA patients, and SMA7 mice were used. Results indicated that LWDH, composed of dihuang, shanyao, shanzhuyu, zexie, hoelen and mudanpi, attenuated LPS-induced microglial inflammation, MPP+-induced oxidative stress and neurotoxicity, and processed dopaminergic neurons protection and locomotor activity improvement in PD mice. In SMA disease cellular and animal models, LWDH up-regulated SMN protein expression, protected motor neurons against SMN deficiency-induced neuronal damage, and improved muscle strength, body weight and survival of SMA mice. We also evaluated the protective effects and mechanisms of loganin, an iridoid glycoside isolated from Corni Fructus, by our drug developing platform. Results indicated that loganin attenuated MPP+-induced neuronal damage by activating glucagon-like peptide-1 receptor (GLP-1R). In cellular models of SMA, loganin increased SMN protein expression, and protected against SMN deficiency-induced motor neuronal damage via activating insulin-like growth factor-1 receptor (IGF-1R). In disease animal model of SMA, loganin promoted protein synthesis pathway, improved muscle strength, body weight, and survival of SMA mice. Moreover, we also evaluated the protective effects of a phenolic natural product from Moutan Cortex, paeonol, in inflammatory and oxidative cellular models. Results indicated that paeonol attenuated LPS-induced microglial inflammation, and attenuated 6-OHDA- and microglial inflammation-induced neurotoxicity. The present thesis setups a drug developing platform of PD and SMA, and demonstrates the potential of LWDH, loganin, and paeonol on complementary therapy of these diseases. We also suggest the possibility of loganin on the application of muscular atrophy related to protein synthesis pathway impairment. Based on the present thesis, we hope to provide neuroprotective evidence and clinical application of herbal medicine, and contribute to herbal medicine development by using this developed platform of PD and SMA in the future.

並列關鍵字

Herbal medicine ； neurodegenerative diseases ； inflammation ； oxidative stress ； Parkinson’s disease ； spinal muscular atrophy

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