Chia Nan University of Pharmacy & Science Institutional Repository:Item 310902800/24651
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    標題: 研究肝硬化病人血中基質金屬蛋白酶之濃度
    Investigation of the relationship between serum levels of matrix metalloproteinases in cirrhosis patients with the gender and cirrhosis etiology
    作者: 劉志龍
    貢獻者: 嘉南藥理科技大學：生物科技系暨研究所
    黃志揚
    葉東柏
    關鍵字: 肝硬化
    明膠酵素圖譜法
    酵素連結免疫吸附分析法
    C型肝炎
    B型肝炎
    基質金屬蛋白\酶\
    酒精性肝炎
    Cirrhosis
    HBV
    ELISA
    MMPs
    Gelatin zymography
    HC
    Alcoholic
    日期: 2009
    上傳時間: 2011-10-27 14:43:12 (UTC+8)
    摘要: 從解剖學上的角度來觀之，肝臟是聯繫身體各個系統的器官，同時肝
    臟具備多項生化弁遄A因此在代謝上扮演很重要角色。研究顯示造成肝臟
    發炎，進而成為肝硬化的原因有B 型肝炎(HBV)、C 型肝炎(HCV)以及酒精
    性肝炎。有許\多生物標記已顯示與肝臟的硬化以及癌症的生長、侵襲與轉
    移有關， 像是參與細胞外基質分解之基質金屬蛋白\酶\(matrix
    metalloproteinases, MMPs) (包括MMP-1 至 MMP-20 等)等。因此分析參與
    疾病形成過程中的生物標記是相當重要的，並具有潛在的意義。
    本研究中，我們收集健康人(男性22 個/女性8 個)及肝硬化病患之血液
    檢體各30 個。後者包括酒精性肝硬化男性7 個、HＢV 引起的肝硬化10 個
    (男性6 個/女性4 個)及HＣV 引起的肝硬化13 個(男性9 個/女性4 個)。使
    用明膠酵素圖譜法 (Gelatin zymography) 做初步篩選，並了解明膠\酶\
    （ gelatinase ） 存在型式之分怖； 再以酵素連結免疫吸附分析法
    (Enzyme-linked immuno-sorbent assay; ELISA) 測定在血漿樣品中的總
    MMP-9 及總MMP-2 濃度，最後利用T test 去做統計分析，以比較健康人與
    不同致病原因之肝硬化患者間MMP-9 及MMP-2 總濃度的差異。
    本研究顯示明膠\酶\，不論是 MMP-9或MMP-2 均以\酶\原型式（pro-form）
    存在為主。而健康人與不同致病原因之肝硬化患者間總MMP-9 或MMP-2
    濃度差異性的比較結果如下：一、在HBV 引起的肝硬化方面：(1)男性健康
    II
    人的總MMP-9 濃度明顯高於HBV 合併肝硬化的男性病人（P
    =0.025），但在女性健康人的總MMP-9 濃度與HBV 合併肝硬化的女性病
    人的總MMP-9 濃度則無明顯差異（P =0.088）；(2) 不同性別HBV 合併肝
    硬化病人間的總MMP-9 濃度並無明顯差異（P =0.837）；(3)相同性別健康
    人與HBV合併肝硬化病人的總MMP-2 濃度間並無明顯差異（男性P =0.083 ;
    女性P =0.298）；(4) 不同性別HBV 合併肝硬化病人間的總MMP-2 濃度亦
    無明顯差異（P =0.475）。二、在HCV 引起的肝硬化方面：(1)男性健康人
    的總MMP-9 濃度與HCV合併肝硬化的男性病人的總MMP-9 濃度無明顯差
    異（P =0.394），在女性健康人的總MMP-9 濃度與HCV 合併肝硬化的女
    性病人的總MMP-9 濃度亦無明顯差異（P =0.12）；(2) 不同性別HBV 合
    併肝硬化病人間的總MMP-9 濃度並無明顯差異（P =0.219）；(3)男性病人
    的總MMP-2 濃度明顯高於男性健康人的總MMP-2 濃度（P =0.012），但在
    女性健康人的總MMP-2 濃度與HCV 合併肝硬化的女性病人的總MMP-2
    濃度無明顯差異（P =0.068）；(4) 不同性別HBV 合併肝硬化病人間的總
    MMP-2 濃度亦無明顯差異（P =0.171）；三、在酒精性肝硬化方面：(1)男
    性健康人的總MMP-9 濃度與酒精性肝硬化的男性病人的總MMP-9 濃度無
    明顯差異（P =0.489）；(2)酒精性肝硬化的男性病人的總MMP-2 濃度較高
    於健康人的總MMP-2 濃度（P <0.001）。
    綜合以上的結果可以得到如下的結論：（1）健康男性的總MMP-9 濃
    III
    度明顯高於HBV 引起的肝硬化的男性病人的總MMP-9 濃度（P =0.025）；
    酒精性肝硬化的男性病人的總MMP-2 濃度明顯高於健康男性的總MMP-2
    濃度（P =0.025）；HCV 引起的肝硬化的男性病人的總MMP-2 濃度明顯高
    於健康男性的總MMP-2 濃度（P =0.012）；（2）在不同原因引起的肝硬化
    及病人之間的比較：健康人(男性及女性)血漿中的總MMP-9 濃度高於HBV
    引起的肝硬化病人(男性及女性) 的總MMP-9 濃度 （P < 0.001）及HCV 引
    起的肝硬化病人(男性及女性) 的總MMP-9 濃度（P < 0.001）。酒精肝硬化
    男性病人血漿中的總MMP-9 濃度高於HBV 引起的肝硬化病人(男性及女性)
    的總MMP-9 濃度（P =0.029）及HCV 引起的肝硬化病人(男性及女性) 的
    總MMP-9 濃度（P = 0.052）。健康人(男性及女性)血漿中的總MMP-2 濃
    度低於HBV 引起的肝硬化病人(男性及女性)的總MMP-2 濃度（P =0.009）、
    HCV 引起的肝硬化病人(男性及女性)的總MMP-2 濃度（P < 0.001）和酒精
    肝硬化男性病人的總MMP-2 濃度（P =0.001）。再者，酒精肝硬化男性病
    人血漿中的總MMP-2 濃度高於HBV 引起的肝硬化病人(男性及女性)（P
    =0.004）血漿中的總MMP-2 濃度。HCV 引起的肝硬化病人(男性及女性)
    血漿中的總MMP-2 濃度高於HBV 引起的肝硬化病人(男性及女性) （P
    =0.02）
    以上數據顯示，不論MMP-9 或MMP-2 在肝硬化病人血漿中的含量的
    確會有所變化，這種現象對於臨床上肝硬化病人之病情診斷分析與治療或
    IV
    預防上將可提供相當程度的參考價值。
    The liver is one of vital organ in human being, it has wide range of functions. This organ plays an important role in metabolism and has a number of biochemical functions. The past studies had shown hepatic disease mainly caused by hepatitis B virus (HBV), hepatitis C virus (HCV) and alcoholism may eventually lead to liver cirrhosis. Many biomarkers have been shown liver cirrhosis related to hepatocellular carcinoma (HCC), including carcinogenesis, invasiveness and metastasis, such as proteinases are involved in the breakdown of extracellular matrix (e.g., MMP-2, MMP-9). Therefore, biomarkers analysis has been shown to be an effective method of identification and monitoring.
    In this study, we collected blood samples from 30 healthy control (22 male/8 female) and 30 liver cirrhosis patients respectively. In liver cirrhosis groups, including 7 males of alcoholic liver cirrhosis, 10 of HBV-induced liver cirrhosis（6 males/4 females）and 13 of HCV-induced live cirrhosis（9 males/4 females）patients. Firstly. a gelatin zymography was performed for preliminary screen and form distribution of gelatinase. Thereafter, an ELISA (Enzyme-linked immuno-sorbent assay) was used for the determination of total MMP-9 and total MMP-2 concentration in the blood samples. By T-test analysis, the enzyme concentration between normal control and cirrhosis patients with different causation were further compared.
    The results of zymography in this study showed that either MMP-9 or MMP-2, the majority of pro-form of both enzymes were found. The matrix metalloproteinase concentration comparison with ELISA between normal control and cirrhosis patients with different causation were obtained as followed. Firstly; in the HBV-induced liver cirrhosis: (1) In healthy males (P = 0.025), but not in females (p = 0.088), the total MMP-9 concentration were significantly higher than in patients; (2) The total MMP-9 concentration in HBV-induced liver cirrhosis males and females was no significant different (p = 0.837); (3) The total MMP-2 concentration in healthy control and HBV-induced liver cirrhosis of same gender was no significant different (male p = 0.083; female p = 0.298). (4) The total MMP-2 concentration in HBV-induced liver cirrhosis males and females was also no significant different (p = 0.475). Thirdly; In the alcoholic liver cirrhosis: (1) The total MMP-9 concentration in healthy control and alcoholic liver cirrhosis male was no significant different (p = 0.489); (2) The total MMP-2 concentration in males patients were significantly higher than in healthy males (P < 0.001)。
    These results can be concluded as follows: (1) In healthy males total MMP-9 concentration were significantly higher than in males patients (p = 0.025). In alcoholic liver cirrhosis males patients total MMP-2 concentration was significantly higher than in healthy males (p = 0.025). In HCV-induced liver cirrhosis male patients total MMP-2 concentration were significantly higher than in healthy males (p = 0.012); (2) In comparison with three different cirrhosis groups, total MMP-9 concentration in both gender healthy controls were significantly higher than HBV-induced (p < 0.001) and HCV-induced liver cirrhosis patients (p < 0.001). The total MMP-9 concentration in alcoholic liver cirrhosis male patients was significantly higher than in both genders of HBV-induced (p = 0.029) and HCV-induced liver cirrhosis patients (p = 0.052). The total MMP-2 concentration in healthy person was significantly lower than HBV-induced (p = 0.029), HCV-induced (p < 0.001) and alcoholic (p = 0.001) liver cirrhosis patients. Furthermore, the total MMP-2 concentration in alcoholic liver cirrhosis patients was significantly higher than HBV-induced liver cirrhosis patients (p = 0.004) of both genders. The total MMP-2 concentration HCV-induced liver cirrhosis patients were higher than HBV-induced liver cirrhosis patients (p = 0.02).
    The data been presented above indicate that the total concentration of either MMP-9 or MMP-2 in plasma samples of liver cirrhosis patients is specifically changed, these phenomena will be significant in clinically diagnosis and treatment or prevention of liver cirrhosis.
    關聯: 校內外完全公開 ，學年度：97， 82 頁
    顯示於類別:[生物科技系(所)] 博碩士論文

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