建立高通量藥物篩選系統尋找具有抗脂肪肝活性之天然物

非酒精性脂肪肝病包括從單純的肝臟內脂肪堆積(脂肪肝)，到出現細胞壞死及嚴重的肝臟發炎反應並伴隨著肝臟纖維化、肝硬化。脂肪肝可以導致肝纖維化，而慢慢發展為肝硬化，並且在此過程中提高了發生肝臟衰竭及肝癌的風險。在過去幾十年中非酒精性脂肪肝已成為已發展國家和發展中國家最常見的肝臟疾病。由於久坐的生活方式和現代西方飲食，台灣目前的脂肪肝盛行率約在12-37％，並正在迅速增長。儘管最有效的治療方法是透過減重和運動，但輔助性的藥物治療策略仍有臨床上的迫切需求。天然物及其衍生物長久以來被用作治療疾病的藥物來源。天然物結構體具有高度的化學多樣性、生化特異性，使它們常常成為藥物開發的先導結構。在本研究中，我們利用本學院陳益昇教授提供的3,000個台灣產植物萃取物，建立了適用於高通量藥物篩選的天然物藥庫。利用老鼠正常肝臟細胞株AML-12以及脂滴染色法做為藥物篩選平台，我們發現了22個萃取物可以抑制細胞內脂滴的堆積，且有劑量依存關係。我們將針對這些有效萃取物做進一步驗證。在本研究的第二部分中，我們在小鼠模式上測試1,2,3,4,6-五没食子酰葡萄糖(PGG)是否能夠抑制高脂肪飲食所誘導的非酒精性脂肪肝。結果顯示，給與小鼠每公斤體重300毫克的PGG可以緩解因高脂肪飲食所引起的體重增加並降低血清中三酸甘油酯含量。肝臟組織病理學檢驗也發現PGG能有效抑制肝臟中的脂肪堆積。未來將可針對其藥理機轉做進一步的探討。

關鍵字

非酒精性脂肪肝病；高通量藥物篩選；高內涵藥物篩選

並列摘要

Non-alcoholic fatty liver disease (NAFLD) can be considered as a spectrum of liver pathologies including simple steatosis with absence of necrosis or signs of inflammation and severe signs of inflammation with fibrosis or cirrhosis. Hepatic steatosis often leads to liver fibrosis, which can progress to cirrhosis with a high risk of liver failure and hepatocellular carcinoma. NAFLD has become the most frequent liver disease during the past decades in developed and developing countries. Due to sedentary lifestyle and modern western nutrition, the prevalence of NAFLD is now around 12-37% in Taiwan and is growing rapidly. Despite the most efficient treatment is by weight loss and exercise, the adjunctive pharmacological strategies are urgently needed. Natural products and their derivatives have historically been used as a source of therapeutic agents. It has long been recognized that natural product structures have the characteristics of high chemical diversity, biochemical specificity that make them favorable as lead structures for drug discovery. In the present study, we constructed a natural product library containing 3,000 Taiwan indigenous plant extracts (kindly provided by Prof. Ih-Sheng Chen) in the format suitable for high-throughput screening. Normal liver cell line-AML-12 and lipid droplet (LD) staining assay were used for establishing an image-based high-throughput screening platform. We identified 22 extracts that inhibited intracellular LD accumulation in dose-dependent manner. The screening hits will be further validated. In the second part of this study, we tested the effect of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (PGG) on high fat diet (HFD) induced NAFLD mouse model. We found that PGG alleviated HFD induced body weight gain and the elevation of serum TG level. Pathological examination revealed that PGG reduced hepatic lipid accumulation significantly. The mechanism of action will be elucidated in the near future.

並列關鍵字

NAFLD ； HTS ； HCS

參考文獻

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建立高通量藥物篩選系統尋找具有抗脂肪肝活性之天然物

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