中草藥對於亨丁頓舞蹈症模式小鼠的神經保護機制

Abstract

亨丁頓舞蹈症（HD）是一種無法治癒的神經退化性疾病，儘管多年的研究，關於亨丁頓舞蹈症神經退化病變的機制仍然不清楚。中國傳統醫藥可以提供新的見解或HD的新療法。中草藥配方的B401是一個眾所周知的台美專利配方，由六種中草藥成分組成。本論文研究主要探討中草藥配方B401對於亨丁頓舞蹈症的神經保護作用。本實驗比較口服中草藥配方B401實驗組以及控制組之R6 / 2小鼠的壽命和體重；透過行為實驗比較口服中草藥配方B401對R6 / 2小鼠的改善情形；利用moorFLPI雷射都普勒成像儀測量R6 / 2小鼠的皮下微循環；利用核磁共振成像技術測量R6 / 2小鼠的腦部體積變化；利用化學發光方法測量R6 / 2小鼠血液中的反應性氧物質（ROS）；利用免疫組織化學染色和西方轉漬技術分析10週齡R6 / 2小鼠腦組織中的關於神經保護、血管生成、氧化壓力，發炎反應和細胞凋亡相關的蛋白質表現，我們的研究結果顯示R6 / 2小鼠口服B401後會延長生存時間、降低體重損失、並且提高運動能力。此外，B401處理也會增強R6 / 2小鼠皮下的微循環；緩解大腦、中腦和小腦的萎縮；並且降低血液中的ROS生成。從我們的研究顯示，口服B401治療顯著增加R6 / 2小鼠腦組織當中的BDNF、VEGF、以及SOD2的表現；但是降低亨丁頓蛋白聚集以及TNF-α的表現。此外，口服B401治療也顯著增加R6 / 2小鼠腦中Bcl-2的表現，但是顯著降低BAX、Calpain、以及Caspase 3的表現。我們的研究顯示口服B401治療亨丁頓舞蹈症R6 / 2小鼠的神經保護機制主要是通過增強神經與血管的新生，但是抑制興奮性毒殺，氧化壓力，發炎反應與細胞凋亡。因此我們認為中草藥配方B401應該可以開發成為改善亨丁頓舞蹈症神經退化性疾病的有效保健品。Huntington’s disease (HD) is a neurodegenerative disease characterized by motor dysfunction, and early death. Despite years of research, the mechanisms responsible for chronic neurodegeneration of HD remain elusive. Chinese traditional medicines might provide new insights or new therapy for HD. The Chinese herbal formula B401 is a well-known Taiwan-US patent formula and consists of six herbal ingredients. My study aimed to elucidate neuroprotective effects of the Chinese herbal formula B401 in the syndrome of HD. We compared the lifespan and body weight of R6/2 mice with and without oral B401 treatment. The ameliorative effects of B401 on symptom of HD mice were investigated through behavior tests. We measured the microcirculation of R6/2 mice by using moorFLPI Laser Doppler Imager, the brain volume of R6/2 mice by using magnetic resonance imaging, the reactive oxygen species (ROS) levels in the blood of R6/2 mice by using chemiluminescence methods, and expressions of proteins for neuroprotection, angiogenesis, oxidative stress, inflammation, and apoptosis in the brain tissue of 10-week-old R6/2 mice by using immunohistochemical staining and western blotting analysis. Our results showed oral B401 treatment increases duration of survival, reduces loss of body weight, and improves motor ability. In addition, B401 treatment enhances subcutaneous microcirculation, alleviates atrophy of cerebrum, midbrain and cerebellum, and reduces blood ROS of R6/2 mice. Evidences from our study show that oral B401 treatment significantly increases the brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), and superoxide dismutase 2 (SOD2), but reduces the huntingtin and tumor necrosis factor-alpha (TNF-α) levels in the brain of R6/2 mice. Furthermore, oral B401 treatment significantly increases B-cell lymphoma 2 (Bcl-2), but reduces Bcl-2-associated X protein (Bax), calpain and caspases-3 in the brain of R6/2 mice. Our study demonstrated the neuroprotection of oral B401 treatment within the brain tissue of R6/2 mice is via enhancing neurogenesis and angiogenesis, but suppressing excitotoxicity, oxidative stress, inflammation and apoptosis. We suggest that the herbal formula B401 can be developed as a potential health supplement for ameliorating the neurodegenerative symptoms of HD.