利用高內涵細胞影像分析儀篩選調控脂肪代謝之肝臟保護藥物

Novel Potential Drugs Identification for Liver Lipid Metabolism Modulation by High Content Screening

鄭婷羽(Ting-Yu Cheng)

指導教授 ： 蘇剛毅

國立臺灣大學/醫學院/醫學檢驗暨生物技術學研究所/碩士(2015年)

https://doi.org/10.6342/NTU.2015.02407

摘要

肥胖為近年來常見且影響健康的問題，容易伴隨著許多疾病的發生，例如糖尿病、心血管疾病以及代謝疾病；而肥胖的形成，少數人是基因所導致，然而大部分的形成可歸因於現代人生活型態的改變，攝取過多養分且缺乏運動所造成。此外，亦有研究指出，肥胖在多種癌症扮演著關鍵的角色，肥胖可使罹患肝癌的機率上升4.5倍，亦參與大腸直腸癌、乳癌、舌咽癌、腎臟癌、胰臟癌等多種癌症的發生。因此，如何有效預防肥胖的產生以及後續治療成為當今重要的議題，其可是透過飲食調控、運動方式，甚至是利用藥物控制體內脂肪的代謝。因此，為了找出能夠有效調節肝臟脂肪代謝的新穎性潛力藥物，本實驗結合高內涵影像分析儀(High content screening)系統、Library of Pharmacologically Active Compounds (LOPAC)藥物化合物以及高脂性細胞培養環境應用於篩選可明顯調控脂肪代謝的新藥。在篩選過程中，培養於高脂性環境下之SK-hep1細胞，其細胞內平均油滴數目為51.8±18.7個，相對於正常環境下之細胞內平均油滴數為8.9±3.5，而在LOPAC1284個藥物化合物中，其中能使細胞內油滴數目相較於未加藥對照組減少超過50%，且細胞存活率達50%以上有66個，細胞存活率達80%以上有30個。而後本實驗利用老鼠初代肝臟細胞做確認，一共挑出五個藥物化合物，且五個藥物對於細胞皆無明顯毒性，而接下來的實驗將著重於探討藥物影響機制以及利用動物模型作確認，篩選出之藥物可供了解脂肪肝相關疾病致病機制，亦可對於肝臟疾病預防治療提供保健策略。

關鍵字

肥胖 ； 脂肪肝 ； 高內涵影像分析儀 ； LOPAC1280

並列摘要

Obesity, a common health condition, is also associated with many clinical disease including diabetes, cardiovascular disease and metabolic syndrome. The occurrence may be due to changes in lifestyle introduced in the 21st century which comprise increased consumption of energy dense foods and reduced physical activity. In addition, obesity plays an important role in other disease progression such as hepatocellular carcinoma (HCC) (risk by up to 4.5-fold), colorectal cancer, breast cancer, esophagus cancer, kidney cancer, pancreatic cancer and leukemia. Therefore drugs or diet supplements for metabolic modulation especially lipid biosynthesis may provide a disease prevention or a therapeutic strategy. In order to develop potential novel drug for this issue, we combined high content screening system, LOPAC1280(Library of Pharmacologically Active Compounds) and high fat medium culture system to screening potential drugs which can significantly modulate lipid metabolism or reduce the accumulation of cellular lipid droplets. Up to date, we have already finished screening all of drug library. We found SK-hep1 cells treated with high fat medium exhibited around 51.8±18.7 lipid droplets per cell compared to normal growth medium treatment (8.9±3.5 lipid droplets per cell). Among 1284 screened drug compounds, about 2.3% of them could effectively decrease more than 50% lipid droplet number compared with cells without drug treatment. We further identified 5 compounds that could reduce the accumulation of intracellular lipid droplets by utilizing mice primary hepatocyte culture system. These compounds also showed no cytotoxicity to hepatocyte in MTT assay. In the furture, these drugs need to be further investigated by experimental animal model as well as the underlying mechanism. The drug candidates may not only have benefits for liver protection but also provide a strategy for fatty liver prevention and therapy in health management.

並列關鍵字

Obesity ； Nonalcoholic Fatty Liver Disease ； High content screening ； LOPAC1280

參考文獻

1. The burden of overweight and obesity in the Asia-Pacific region. Obes Rev, 2007. 8(3): p. 191-6.

2. Chu, N.F., Prevalence of obesity in Taiwan. Obesity Reviews, 2005. 6(4): p. 271-274.

4. Mitchell, S. and D. Shaw, The worldwide epidemic of female obesity. Best Practice & Research Clinical Obstetrics & Gynaecology, (0).

6. Ferre, P. and F. Foufelle, Hepatic steatosis: a role for de novo lipogenesis and the transcription factor SREBP-1c. Diabetes Obes Metab, 2010. 12 Suppl 2: p. 83-92.

7. Bechmann, L.P., et al., The interaction of hepatic lipid and glucose metabolism in liver diseases. J Hepatol, 2012. 56(4): p. 952-64.