慢性酒精攝取下野生牛樟芝對肝臟保護作用的探討

Hepatoprotection of wild fruiting body of Niuchangchih(Antrodia camphorata) in chronic alcohol consumption.

康琝毓(Wen-Yu Kang)

指導教授 ： 陳億乘

中山醫學大學/健康管理學院/營養學研究所/碩士(2010年)

https://doi.org/10.6834/CSMU.2010.00026

摘要

長期飲酒導致酒精性脂肪肝，進而引起肝發炎、肝硬化。在原住民口耳相傳下，認為牛樟芝(Antrodia camphorate, AC)可以解宿醉，改善肝疾病等病症等等的作用，然而市面上牛樟芝多為人工培養對於保護肝臟的效用眾說紛紜，因此本實驗為探討野生牛樟芝在慢性酒精攝取下是否具保護肝臟作用﹖本研究挑選五週齡雄性Wistar大鼠將其分為四組，飲食皆為chow diet，飲水以20%酒精水溶液取代，先飼養一週適應環境，一週後隨機分組成四組，每組十隻大鼠，分組如下：一) ALC組：給予20%酒精水溶液；二) ALC_Sil組：給予20%酒精水溶液＋0.25g/kg b.w.劑量的silymarin；三) ALC_1X組：給予20%酒精水溶液＋0.025g/kg b.w.劑量的野生牛樟芝子實體粗萃取物；四) ALC_4X組：給予20%酒精水溶液＋0.1g/kg b.w.劑量的野生牛樟芝子實體粗萃取物，持續飼養4週後發現ALC_4X組具有較低的內臟脂肪的重量百分比(p＜0.05)，野生牛樟芝子實體粗萃取物組別(ALC_1X組，ALC_4X組)有較低血清中TC的濃度(p＜0.05)，並且有較少的肝臟中脂肪含量以及TC與TAG (只有ALC_4X組)濃度(p＜0.05)，較高的糞便中TC與膽酸的排泄(p＜0.05)。ALC_1X組與ALC_4X組中肝臟破損指數(AST、ALP)也比ALC組低。另外，野生牛樟芝子實體粗萃取物也可以降低慢性酒精攝取下血清與肝臟中脂質過氧化物(MDA)濃度(p＜0.05)，提高肝臟中總抗氧化能力(TEAC) (p＜0.05)，而肝臟中GSH含量只有ALC_Sil組與ALC_4X組顯著增加(p＜0.05)。酒精代謝方面：ALC_1X組與ALC_4X組中肝臟中ADH、ALDH的基因表現量上升(p＜0.05)，另外，野生牛樟芝子實體粗萃取物也可提升ADH (只有ALC_4X組)、ALDH酵素活性(p＜0.05)，降低血清中酒精濃度(p＜0.05)。在代謝酒精的另一條路徑中，雖然0.1g/kg b.w.野生牛樟芝劑量會向下調控肝臟的CYP2E1的基因表現量(p＜0.05)，但是也減少其代謝過程中所產生的自由基，降低肝臟氧化壓力。同時，野生牛樟芝子實體粗萃取物也幫助提高肝臟中抗氧化酵素SOD、CAT與GSH-Px的活性(p＜0.05)，清除體內因代謝酒精產生的過多自由基，降低肝臟受傷機會，而CAT活性的提高也加速酒精的代謝。在體內膽固醇恆定(Cholesterol homeostatsis)相關基因表現：ALC_1X組與ALC_4X組可降低肝臟中HMG-CoAR基因表現(p＜0.05)減少內生性膽固醇的合成。在體內脂肪酸與三酸甘油酯(Fatty acid and triacylglycerol)生成相關基因表現：野生牛樟芝子實體粗萃取物可降低肝臟中SREBP-1c、ACC (只有ALC_4X組)與FAS基因表現(p＜0.05)。在能量代謝(Energy metabolism)相關基因表現：ALC_4X組中肝臟中PPAR-α的基因表現量相對提高(p＜0.05)。從肝臟切片中觀察慢性飲酒大鼠補充野生牛樟芝子實體粗萃取物可以減少肝臟細胞壞死的病灶與發炎現象產生，同樣在與肝臟發炎相關的基因表現中發現，野生牛樟芝子實體粗萃取物可以降低TNF-α、KLF-6與TGF-β1的基因表現，但是下降的AP-1基因表現只有在0.1g/kg b.w.劑量下發現(p＜0.05)。本實驗結果指出：在慢性酒精攝取下，野生牛樟芝子實體粗萃取物具有降低血清總膽固醇濃度、肝受損指數與肝臟脂肪含量，促進糞便總膽固醇與膽酸的排泄，提高肝臟抗氧化能力，避免酒精氧化造成的傷害；另外，野生牛樟芝子實體粗萃取物則可抑制因慢性酒精攝取下所造成的脂質生成以及發炎相關的基因表現；此外，野生牛樟芝子實體粗萃取物也可相對提升能量代謝相關基因表現，繼而降低肝代謝異常所造成的傷害。

關鍵字

酒精 ； 牛樟芝 ； 脂肪肝

並列摘要

A chronic alcohol comsumption can cause alcoholic fatty liver (AFL), further leading to hepatitis, fibrosis, and cirrhosis. Commercially, Antrodia camphorata (AC) is announced a hepatoprotection. Based on our knowledge, a comprehensive report on the wild AC hepatoprotection against a chronic alcohol comsumption is not available. Hence, this study would like to investigate the wild AC hepatoprotection against a chronic alcohol comsumption via regulations of lipid homeostasis, alcohol metabolism, antioxidant capacity, and profibrosis/inflamation in a rat model.
After one week accumulation, 40 male Wistar rats (5-wk age) all drank 20% alcohol solution ad libitum and assigned randomnly into four groups: 1) no treatment (ALC); 2) 0.25g silymarin /kg b.w./day (ALC_Sil); 3) 0.025g wild AC /kg b.w./day (ALC_1X); 4) 0.1g wild AC /kg b.w./day (ALC_4X). The experiment lasted for 4 weeks.
Results indicated that lower (p<0.05) serum TC level, and hepatic TC and TAG levels were observed in the wild AC supplemented groups, especially the ALC_4X group, compaired to those of the ALC group. Meanwhile, wild AC supplemented groups could increase (p<0.05) the TC and bile acid excretion in feces. In regulation of lipid homeostasis in liver of alcohol-fed rats, wild AC downregulated (p<0.05) HMG-CoAR, SREBP-1c, ACC, and FAS gene expressions; while an upregulation (p<0.05) of PPAR-α gene expression was only observered in the ALC_4X group.
Although CYP2E1 gene expression in alcohol-fed rats was downregulated (p<0.05) by supplementing 0.1g wild AC/kg b.w./day, wild AC upregulated (p<0.05) hepatic ADH and ALDH gene expressions, and meanwhile enhanced (p<0.05) the hepatic ADH and ALDH activities, which lower (p<0.05) the serum alcohol concentration. The wild AC also increased (p<0.05) the liver GSH and TEAC levels, as well as, SOD, CAT, and GSH-Px activities, and meanwhile decreased (p<0.05) MDA levels. Besides, the increased CAT activity by supplementing wild AC to alcohol-fed rats could accelerate alcohol metabolism. The wild AC also downregulated (p<0.05) the hepatic TNF-α, KLF-6, AP-1, and TGF-β1 gene expressions of alcohol-fed rats. Following above benefits on the wild AC against a chronic alcohol comsumption, lower (p<0.05) AST and ALP values, as well as mild or no liver necrosis, inflammation and infiltration via a histological examination were observed.

並列關鍵字

alcohol ； Antrodia camphorata ； fatty liver

參考文獻

陳勁初、呂鋒洲，2001年，靈芝之王：台灣樟芝。

彰化基督教醫院，2010年，消化系中心，http://www2.cch.org.tw/gi/index.htm。

Agarwal, D. P. (2001). Genetic polymorphisms of alcohol metabolizing enzymes. Pathologie Biologie 49, 703-709

Aksenov, M. Y. and Markesbery, W. R. (2001). Changes in thiol content and expression of glutathione redox system genes in the hippocampus and cerebellum in Alzheimer’s disease. Neuroscience Letters 302,141-145.

Allen, J. P., Litten, R. Z., Anton, R. F., and Cross, G. M. (1994). Carbohydrate-deficient transferrin as a measure of immoderate drinking: Remaining issues. Alcoholism, Clinical and Experimental Research 18, 799-812.