應用蛋白質體學研究中藥茵陳蒿湯對保肝作用的分子調控機制(2-2)

李宗諺
長庚大學
膽汁淤積對於肝細胞損傷是一項重要的因素，同時也對肝臟細胞凋亡扮演重要的角色。傳統中醫藥以1) 疏肝解鬱；2) 活血除濕為原則，提供有效與安全的治療。中藥方劑茵陳蒿湯可以改善肝臟膽紅素清除率，改善黃疸現象。臨床文獻報導中國大陸和日本使用中藥方劑茵陳蒿湯治療不同類型的肝疾病，發現其具有利膽與保肝的特性。因此，本計劃藉由膽道阻塞手術的動物模式來探討茵陳蒿湯對於肝臟細胞凋亡與肝纖維化的調控機制。膽道結紮手術使大鼠因膽汁淤積產生肝纖維化。術後實驗動物在清醒狀態下，連續27天給予茵陳蒿湯(50、 125、250 mg/kg BW)或生理食鹽水的胃管餵食。第28天，動物在麻醉狀態下收取血清做游離脂肪酸、膽固醇、三酸甘油脂、與腫瘤壞死因子(TNF-a)的分析。所有實驗動物皆收取肝臟組織，部分作為病理組織切片，部分則以西方印漬法分析細胞凋亡相關蛋白做半定量的測定。反轉錄多鏈聚合酶反應分析肝臟組織中過氧小體增生活化受體-g，a，d與固醇調節因子結合蛋白 mRNA的表現。另外，為了進一步瞭解茵陳蒿湯對交互作用的蛋白質網路完整的病理機制的調控，本實驗也將以蛋白質體學(proteomics)對檢體進行研究。本實驗的執行，我們藉由探討茵陳蒿湯對膽汁鬱滯所引起肝細胞凋亡的分子機制調控，同時對茵陳蒿湯在肝纖維化治療過程中肝臟組織、血清表現差異極大的蛋白質作分類，以嘗試建立茵陳蒿湯對肝纖維化的療效機制圖譜。透過更多的實驗證據來加強我們對茵陳蒿湯在膽汁瘀積的療效評估並對慢性肝疾病能有更多科學的思維。
關鍵字：茵陳蒿湯；膽汁淤積；過氧小體增生活化受體

Application of Proteomics on Hepatoprotection Effect of Chinese Herb Medicine Yin-Chen-Hao-Tang(2-2)

Lee, Tzung-Yan
Chang Gung University
The accumulation of hydropholic bile acids in the liver is considered to play a pivotal role in the induction of apoptosis of hepatocyte during cholestasis. Treatment with traditional Chinese medicine drugs has been proved satisfactory and curative effect with principles of (1) soothing the liver and dispelling stasis; (2) promoting blood circulation and excreting dampness. A Chinese herbal remedy called Yin-Chen-Hao-Tang (YCHT or ICHT ) is now shown to attenuate development of hepatic bilirubin clearance. This herbal decoctions have been recognized as a ’’magic bullet” for jaundice and has long been used in China and Japan as a choleretic and hepatoprotective agent for various types of liver diseases. Therefore, we used an experimental animals model of biliary atresia to test the hypothesis that YCHT plays a key regulatory role in the pathogenesis of bile duct obstruction in hepatic apoptosis. Experimental cholestasis-induced liver fibrosis after common bile-duct ligation (BDL) in rats. After BDL surgery, YCHT (50, 125, 250 mg/kg body p.o.) continued for 27 days.
關鍵字：Yin-Chen-Hao-Tang；cholestasis；peroxisome proliferators-activated receptors