應用活體多光子顯微術來探討脂肪肝對肝膽代謝功能的影響

Abstract

肝病是影響人類健康的重要因素之一，而肝癌也一直位居癌症十大死因前一、二名。脂肪肝是肝病的一種，而台灣在患脂肪肝的人口比例上竟然位居世界之首。1脂肪肝的患者如果不做規劃性或藥物性的控制，有很大的機率會惡化成脂肪肝炎或是肝硬化。脂肪肝的重要性吸引了許多學者投入研究，希望能對這個病症的治療或預防有更多的貢獻。 在我的研究之中，我主要結合了多光子顯微術、本實驗室自製的肝觀察視窗平台、與螢光劑6-CFDA來進行活體實驗。非螢光性的6-CFDA會在肝細胞內被分解成發螢光的6-CF，然後被肝細胞的蛋白質運輸出去，我們利用6-CF在肝細胞內的強度變化來估計肝細胞的代謝能力。另外我使用的動物模型是餵食MCD的B57CL/6小鼠，分別在餵食後的第二、四、六、八個禮拜做實驗，實驗結果與控制組老鼠比較顯示患脂肪肝會造成肝表現較差的代謝能力。

Liver disease represent a major health problem worldwide. Fatty liver is a common liver disease which can progress to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis. In this thesis, in order to elucidate the effects of fatty liver on hepatobiliary metabolism, we combined multi-photon microscopy with a home-built intravital hepatic imaging chamber (IHIC) to image and quantify the metabolic capacity of liver in methionine- and choline-deficient (MCD) dietary animal model. Mice were imaged at 2, 4, 6, and 8 weeks after they were first fed with MCD. Non-fluorescent 6-carboxyfluorescein diacetate (6-CFDA) which is hydrolyzed by the hepatocytes into fluorescent 6-carboxyfluorescein (6-CF) and excreted into the bile canaculi by the multidrug resistant proteins (mrps) is then used as the molecular probe in tracing the effects of fatty liver on hepatobiliary metabolism. Our data showed that the metabolic function of liver in fatty liver diseased mice is impaired in comparison with that of normal mice.