利用多光子顯微術觀察活體嚴重脂肪肝細胞代謝行為

非酒精性指肪肝(NAFLD)是現今世上最流行的疾病之一。這種病症被認為是與許多其他的併發症有相關性，其中包括最普遍的脂肪肝、脂肪性肝炎或是肝硬化等。而上述所有併發症的病理前因被認為是與肥胖非常相關的胰島素不耐症。由此可推論在已開發國家的久坐與高醣、高油脂飲食生活習慣中，非酒精性脂肪肝影響的人口勢必日益趨增。在統計上全世界非酒精性脂肪肝的盛行率大約佔了百分之十至二十四的人口。在肥胖與第二型糖尿病族群中則高達百分之九十，其中包含百分之二十的脂肪性肝炎與百分之十的肝硬化患者。又在肝硬化患者中每年有百分之一至二的患者會惡化為肝癌。雖然僅僅是脂肪肝尚未被認為是對人體有害，但可能是某種併發症的前兆。因此早期檢測出脂肪肝或研究出脂肪肝更細微的生理機制將會對臨床醫學有所助益。本論文將探討使用多光子激發螢光技術研究脂肪肝對C57BL/6小鼠活體肝細胞代謝能力的影響。以六週大的小鼠經由蛋胺酸(methionine)與膽鹼(choline)缺乏的飲食所引發出的脂肪肝為模型，進行對6-carboxyfluorescein diacetate (6-CFDA) 的代謝做觀察。實驗結果成功驗證脂肪肝對個體代謝速率有負影響，並且此影響對同一個體內的不同肝細胞會造成不均勻的效果。

關鍵字

非酒精；脂肪肝；代謝；活體；小鼠；多光子

並列摘要

Nonalcoholic fatty liver disease (NAFLD) is one of the most prevalent diseases in the world. This condition refers to wide spectrum of liver disease ranging from the most common fatty liver (fat accumulation in the liver, also known as steatosis), to non-alcoholic steatohepatitis (NASH, fat caused inflammation in liver), to cirrhosis (an irreversible process of the inflammation progressively scarring the liver). All of the above pathological changes are now believed due to insulin resistance, which is closely associated with obesity. Accordingly, NAFLD affects increasingly more people nowadays, especially in developed countries where the sedentary lifestyle and the high calories, sugar and fat diet make the prevalence of obesity, insulin resistance and diabetes. The statistic prevalence of NAFLD in population is 10~24% worldwide. This number approaches 90% in obese and diabetes mellitus type 2 patients, includes 20% with NASH and 10% with cirrhosis. Among those with cirrhosis, primary liver cancer develops at rate of 1~2% per year. However, simple fatty liver itself usually seemed harmless but maybe a forecast of many complications. Therefore, it will be beneficial if fatty liver can be diagnosed earlier and if the mechanisms associated with the pathogenesis of this disease are better known. This thesis utilizes multi-photon microscopy to evaluate how the accumulated fats affect the liver metabolism in C57BL/6 mice in vivo. . 6 weeks old mice were fed with the methionine choline deficient diet (MCD diet) or supplied control diet (MCS diet) ad libitum for up to 8 weeks. The MCD diet can quickly induce fatty liver by impairing the synthesis of very low density lipoprotein (VLDL) in hepatocyte. After the onset of fatty liver, 6-carboxyfluorescein diacetate (6-CFDA) was injected as a molecular probe. When this non-fluorescent compound enters living cells, it is hydrolyzed by esterases, producing the fluorescent compound, 6-carboxyfluorescein (6-CF). Thereafter, the multidrug resistant proteins would drain these fluorescent compounds through bile canaliculi. Therefore, the present research aims to image and quantify the effects on hepatobiliary metabolism caused by the accumulated fat in hepatocyte.

並列關鍵字

NAFLD ； fatty liver ； in vivo ； multi-photon ； metabolism ； mice ； MCD

參考文獻

[1] 賴世偉、廖光福、黃金財；健檢民眾非酒精性脂肪肝盛行率之分析；輔仁醫學期刊，第 3 卷，第 2 期 (2005)

[7] Yuan Liu, Feng-Chieh Li, Hsiao-Chin Chen, Po-shou Chang, Shu-Mei Yang, Hsuan-Shu Lee and Chen-Yuan Dong, "In vivo multiphoton imaging of bile duct ligation", Proc. SPIE 6860, 68600G (2008)

[2] Fatty Liver: Nonalcoholic Fatty Liver Disease (NAFLD) and Nonalcoholic Steatohepatitis (NASH) < http://www.medicinenet.com/fatty_liver/article.htm>

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[5] Max Born, E. W. Principles of Optics, Electromagnetic theory of propagation, interference and diffraction of light. 7th edn, (2005).

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利用多光子顯微術觀察活體嚴重脂肪肝細胞代謝行為

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