本文選題：急性肝損傷 + 四氯化碳��； 參考：《東北農(nóng)業(yè)大學》2012年碩士論文

【摘要】：近年來肝臟疾病對人類的健康和社會的發(fā)展造成嚴重威脅,養(yǎng)殖業(yè)中肝病的發(fā)病率明顯增高,不僅給養(yǎng)殖業(yè)造成嚴重的經(jīng)濟損失,而且通過食物鏈直接或間接對人類健康造成危害,引發(fā)諸多食品安全問題。急性肝損傷是多種肝臟疾病的發(fā)生、發(fā)展及最終走向肝功能衰竭的始動環(huán)節(jié)和共同途徑,建立與人類急性肝損傷發(fā)展病變過程相似的動物模型極為重要。深入研究和探討急性肝損傷的發(fā)病機制以及治療措施,對慢性肝臟疾病的防治具有重要意義。GSTA1是Ⅱ相結合反應同功酶GST的一個亞型,在抗氧化損傷、抗腫瘤方面具有重要作用,并有可能成為藥物研發(fā)重要的新方向。目前關于GSTA1在急性肝損傷中的變化研究較少,尚未見GSTA1在急性肝損傷中的作用研究。明確GSTA1在急性肝損傷中的變化將為GSTA1保護作用及其機制的研究提供理論基礎和試驗依據(jù)。 選用雄性昆明小鼠為實驗動物,研究并明確四氯化碳(CCl4)致小鼠急性化學性肝損傷、醋氨酚(APAP)致小鼠急性藥物性肝損傷、乙醇致小鼠急性酒精性肝損傷的建立方法和評價方法。通過篩選獲得最佳的染毒途徑、劑量和時間,以此為基礎復制三種小鼠急性肝損傷模型。CCl4模型以血清轉氨酶(ALT、AST)、肝組織指標(SOD、MDA、GSH、GSH-Px)和肝臟病理組織學分析進行評價；APAP模型以血清轉氨酶(ALT、AST)、肝組織指標(SOD、MDA、GSH、GSH-Px、NO)和肝臟病理組織學分析進行評價；乙醇模型以血清轉氨酶(ALT、 AST)、肝組織指標(SOD、MDA、GSH、GSH-Px、TG)和肝臟病理組織學分析進行評價。并在成功復制的各模型中,檢測血清和肝臟中GSTA1含量和活性變化情況。GSTA1含量檢測為ELISA原理,采用小鼠GSTA1試劑盒進行。GSTA1活性檢測采用比色法,以NBD-Cl為底物,通過GSTA1所催化的酶促反應生成物NBD-SG的吸光度來計算。血清中酶活性表達為每分鐘每毫升血清中GSH反應生成物的量,肝組織中酶活性表達為每分鐘每毫克胞漿蛋白中GSH反應生成物的量。 研究結果表明： 1.以0.35%CCl4油溶液,按10mL·kg-1劑量灌胃,染毒24h可成功復制CC14致小鼠急性肝損傷模型。模型組血清轉氨酶活性升高,與對照組相比差異極顯著(P0.01),肝組織中各指標變化與對照組相比均差異極顯著(P0.01),病理切片觀察到模型組小鼠肝細胞顆粒變性,胞漿崩解,細胞核濃縮,炎性細胞浸潤。血清中GSTA1含量和活性顯著增加(P0.01),而肝臟中GSTA1含量和活性均顯著降低(P0.01)。 2.以200mg·kg-1劑量APAP灌胃,染毒12h可成功復制小鼠急性肝損傷模型。模型組血清轉氨酶活性升高,與對照組相比差異極顯著(P0.01),肝組織中各指標變化與對照組相比均差異極顯著(P0.01),病理切片觀察到模型組肝組織充血、淤血,炎性細胞浸潤,肝索模糊,胞核固縮。血清中GSTA1含量和活性顯著增加(P0.01),而肝臟中GSTA1含量和活性均顯著降低(P0.01)。 3.以50%乙醇,按14mL·kg-1劑量灌胃,染毒8h可成功復制小鼠急性肝損傷模型。模型組血清轉氨酶活性升高,與對照組相比差異極顯著(P0.01),肝組織中各指標變化與對照組相比均差異極顯著(P0.01),病理切片觀察到模型組小鼠肝組織有圓形空泡、炎性細胞浸潤,肝細胞點狀壞死。血清中GSTA1含量和活性顯著增加(P0.01),而肝臟中GSTA1含量和活性均顯著降低(P0.01)。 本試驗成功復制急性化學性、藥物性、酒精性肝損傷動物模型。在三種急性肝損傷模型中,GSTA1含量和活性在血清中顯著增加(P0.01),且在肝組織中顯著降低(P0.01),表明急性肝損傷時GSTA1由肝臟釋放入血,參與機體抗氧化活動,從而起到保護肝臟的作用。血清中GSTA1的變化表明GSTA1對于急性肝損傷的評價具有重要意義。本研究為繼續(xù)探索保肝藥物對GSTA1調節(jié)作用和GSTA1保護肝臟的分子機制打下堅實基礎并提供可靠理論數(shù)據(jù)。
[Abstract]:In recent years , liver diseases have posed a serious threat to human health and social development , and the incidence of liver diseases in the breeding industry is significantly increased . It is very important not only to cause serious economic losses to the breeding industry , but also to directly or indirectly harm the human health through the food chain .

The acute hepatic injury induced by acute chemical liver injury and alcohol induced acute alcoholic liver injury in mice induced by carbon tetrachloride ( CCl _ 4 ) induced acute hepatic injury in mice was studied by using male Kunming mice as experimental animals .
APAP model was evaluated by serum transaminase ( ALT , AST ) , liver tissue index ( SOD , MDA , GSH , GSH - Px , NO ) and histopathological analysis of liver ;
The content and activity of GSTA1 in serum and liver were evaluated by using serum transaminase ( ALT , AST ) , liver tissue index ( SOD , MDA , GSH , GSH - Px , TG ) and histopathological analysis .

The results show that :

1 . The acute liver injury model of CC14 - induced acute liver injury could be successfully replicated by 10 mL 路 kg - 1 dose of 0 . 35 % CCl 4 oil . Compared with the control group , the activity of serum transaminase increased significantly in the model group ( P0.01 ) . The changes of the indexes in the liver tissues were significantly higher than those in the control group ( P0.01 ) . The content and activity of GSTA1 in the liver were significantly increased ( P0.01 ) , while the content and activity of GSTA1 in the liver decreased significantly ( P0.01 ) .

2 . The acute liver injury model of mice was successfully replicated at 200 mg 路 kg -1 APAP . The serum transaminase activity in the model group was significantly higher than that in the control group ( P0.01 ) . The changes of the indexes in the liver tissues were significantly higher than those in the control group ( P0.01 ) . The content and activity of GSTA1 in the liver were significantly increased ( P0.01 ) , while the content and activity of GSTA1 in the liver decreased significantly ( P0.01 ) .

3 . The model of acute liver injury in mice was successfully reproduced at the dose of 14 mL 路 kg - 1 at 50 % ethanol . The serum transaminase activity in the model group was significantly higher than that in the control group ( P0.01 ) . The changes of the indexes in the liver tissues were significantly higher than those in the control group ( P0.01 ) . The content and activity of GSTA1 in the liver were significantly increased ( P0.01 ) , while the content and activity of GSTA1 in the liver decreased significantly ( P0.01 ) .

This study successfully replicated animal models of acute chemical , drug and alcoholic liver injury . In the three models of acute liver injury , the content and activity of GSTA1 increased significantly in serum ( P0.01 ) .
【學位授予單位】：東北農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R-332

【引證文獻】

相關碩士學位論文 前1條

1 劉穎姝;急性肝損傷早期診斷指標GSTA1的研究及保肝藥物初篩[D];東北農(nóng)業(yè)大學;2013年

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本文編號：2052981