周芬碧Fen-Pi Chou廖婉琦Wan-Chi生化暨生物科技研究所2026-10-289999-12-312012https://handle.ncl.edu.tw/11296/jn3wxkhttps://scholars.csmu.edu.tw/handle/123456789/35763肝臟在人體中參與許多生理作用，在人體中為重要器官。過去文獻顯示，荖葉具有抗氧化性，其可透過多條訊息傳遞路徑使體內matrix metalloproteinase-2 (MMP2)蛋白活化去降解alpha – smooth muscle actin (α-sma)沉積造成的肝臟纖維化現象，並且活化體內superoxide dismutase (SOD) 、catalase (CAT)等酵素活性，以證實其保肝之效果。為了更進一步瞭解荖葉之保肝作用是否透過肝臟解毒酵素，本論文使用AAF誘導大鼠肝損傷模式，並於誘導前先行餵食荖葉水萃取物，而後荖葉水萃取物與AAF並行使用。動物犧牲後，以定量PCR測定大鼠肝臟中解毒酵素mRNA之表現量，並且觀看基因表現量與血液中所測得的生化指標兩者之間的關係。結果發現雖然在單獨餵食荖葉水萃取物時並不影響肝臟功能，然而在單獨AAF誘導組中則有13個基因於統計上有著顯著差異，其中CYP1A1、GSTM5、GSTP1三者表現為增加，其餘為明顯減少。且於正常控制組的相關性分析中可以發現P450活性、淋巴球含量以及血小板含量三者與大多數基因呈現負相關之狀態，然而於AAF誘導組中則全數轉變為正相關性之表現。接著在AAF誘導肝損傷之情況中配合餵食荖葉，其可觀察到大鼠肝臟會隨著荖葉濃度之提升，亦隨之增加肝臟結節的病變及惡化的狀況，但在基因定量中並無發現因為荖葉之因素而改變結果，除了極少數基因，如：CYP1A1、CYP2B3、CYP2E1、CYP3A23/3A1等，但這些基因亦無隨荖葉濃度變化而改變，以及血液值相關性分析亦無隨荖葉濃度而變化之規則。由於實驗結果發現解毒酵素並無因為荖葉而影響表現量，表示荖葉的保肝作用可能並不是透過解毒代謝路徑。然而因本研究之餵食時間較長，無法觀察到荖葉之立即影響，不排除動物犧牲時荖葉對解毒酵素基因之作用已回復穩定狀態。Liver is an important organs involved in many physiological roles. Recent research indicates that water extracts of piper betel leaf (PBL) have antioxidant effect. PBL decreased the AST, ALT, and increased SOD, CAT enzyme activities to decrease free radicals induced by CCl4 in rat liver. This effect was elicited via the Ras/Raf signal pathway and induction of TIMP2 to active MMP2 enzyme expression that might enhance the degradation of α-sma, and to ease liver fibrosis. To further understand the hepatoprotective effect of the PBLs, this study use the AAF-induced liver damage model to examine the influence of PBL on the expression of liver detoxification enzymes. Animals were pretreated with PBLs before the induction of AAF, and then PBLs and AAF were given simultaneously. At the end of the study RT-PCR was used to determine the mRNA expression of detoxification enzymes in rat liver that was further correlated to the biochemical markers in sera. The results showed that PBLs alone did not affect liver function. However, AAF influenced the levels of 13 genes in which CYP1A1, GSTM5, and GSTP1 were increased and the rest were decreased. The correlation analysis of the normal group showed that the P450 activity, lymphocyte and platelet were negatively correlated, and became positively correlated in the AAF-induction group. The AAF combined with PBLs group found dose-dependent increases in hepatic nodular lesions and deterioration. However, the combined treatment of PBL did not change the gene expression of AAF, except CYP1A1、CYP2B3、CYP2E1、CYP3A23/3A1. The results show that PBLs did not affect the expression of the detoxification enzymes, indicating that the protective role of PBL may not be through the detoxification metabolic pathway. However, we can not rule out the possibility that the long-term feeding of this study might diminish the immediate impact of betel leaf.thesis