2023-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/648338脂肪肝 (fatty liver disease)是全世界最盛行的慢性肝病之一，當脂肪在肝臟過度累積達5%，就會形成脂肪肝。酗酒是先進國家相當嚴重的社會問題，飲酒過量會促使肝細胞合成的脂肪增加，因此酒精性脂肪肝病(alcoholic fatty liver disease, AFLD)是需要被重視的問題。在AFLD病程中，過量攝取酒精會使腸道通透性增加，腸源性內毒素「脂多醣（Lipopolysaccharide, LPS）」因此洩漏至血液循環中，LPS會被細胞上的Toll-like receptor 4（TLR-4）以及Cluster of differentiation 14（CD14）接收，透過一連串下游調控因子，引起發炎反應。而AFLD會使肝臟結構改變，當肝細胞再生無法填補細胞死亡的空缺時，便產生酒精性肝纖維化，導致胞外基質（extracellular matrix, ECM）過度累積。基質金屬蛋白酶第九型(matrix metalloproteinase 9, MMP-9) 是負責調節ECM持衡的關鍵酵素，因此MMP-9可能參與在酒精性脂肪肝病程中。本研究將以MMP-9基因剔除 (MMP-9 knockout, MMP-9 KO)小鼠餵飼液態酒精飼糧誘發脂肪肝病，再外源注射LPS，比較其與野生型小鼠發炎相關因子TNFα、IL-6及IL-1β表現量、脂質浸潤程度及MMP-9在肝臟之蛋白質表現量與酵素活性。細胞實驗部分將小鼠肝癌細胞株Hepa1-6和小鼠巨噬細胞RAW264.7共培養，並以酒精及LPS處理，評估細胞膜上TLR-4與CD14受體總量，藉此模擬肝臟巨噬細胞在AFLD下的反應，釐清MMP9在酒精性脂肪肝病程詳細生理疾病機制，為未來酒精性脂肪肝病之治療提供更多的可能性。 Fatty liver disease is one of the world`s most prevalent chronic liver diseases and occurs when fat excess accumulates of 5% in the liver. Alcoholism is a very serious social problem in advanced countries. Excessive drinking will increase the synthesis of fat in liver cells. Therefore, alcoholic fatty liver disease (AFLD) needs to be taken seriously. During the course of AFLD, excessive intake of alcohol will increase intestinal permeability, and the intestinal endotoxin "Lipopolysaccharide (LPS)" will leak into the blood circulation. LPS will be received by Toll-like receptor 4 (TLR-4) and Cluster of differentiation 14 (CD14) on cells, and through a series of downstream regulatory factors, causing inflammation. AFLD changes the liver structure, and when liver cell regeneration fails to fill the vacancy of cell death, alcoholic liver fibrosis occurs, resulting in excessive accumulation of extracellular matrix (ECM). Matrix metalloproteinase type 9 (MMP-9) is a key enzyme responsible for regulating ECM homeostasis, for reason that MMP-9 may be involved in the course of AFLD. In this study, we induce fatty liver disease by feeding MMP-9 knockout (MMP-9 KO) mice liquid alcohol diets, and then exogenously injected with LPS. Compare the expressions of TNFα, IL-6, and IL-1β, the degree of lipid infiltration, and the protein expression, and enzyme activity of MMP-9 in the liver with wild-type mice. In the cell experiment part, will co-cultured the mouse liver cancer cell line Hepa1-6 and the mouse macrophage RAW264.7, then treated with alcohol and LPS to evaluate the total amount of TLR-4 and CD14 receptors on the cell membrane.Simulate the response of liver macrophages under AFLD, clarify the detailed physiological disease mechanism of MMP-9 in the course of AFLD, and provide more possibilities for the treatment of AFLD in the future.酒精性脂肪肝;基質金屬蛋白酶第九型;胞外基質;發炎反應;慢性肝病;Alcoholic Fatty Liver Disease;Matrix Metalloproteinase-9;Extracellular Matrix;Inflammation;Chronic Liver Disease