本文選題：氫氣 + 飽和��； 參考：《瀘州醫(yī)學院》2014年碩士論文

【摘要】：目的：本實驗通過建立大鼠肝內膽管缺血再灌注損傷(IR)動物模型，探討靜脈注射氫氣飽和生理鹽水是否對大鼠肝內膽管缺血再灌注損傷具有保護作用，觀察其病理學改變，并從膽管上皮細胞Bcl-2/Fas基因表達、凋亡等方面探討氫氣飽和生理鹽水可能的作用機制。方法：將32只雄性SD大鼠隨機分成4組：假手術組（SO組），缺血再灌注組（IR組），缺血再灌注+生理鹽水組（IR+NS組），缺血再灌注+氫氣飽和生理鹽水組（IR+HS組），每各8只。用1％戊巴比妥鈉以每100g體重0.6ml的劑量注射進入腹腔，等待麻醉藥物效果顯效后，用碘伏對大鼠的腹部進行消毒，然后沿著大鼠的腹正中線打開腹部，分離至膽總管及十二指腸，并于二者匯合處經十二指腸外用硬膜外導管逆行置管于膽總管內，固定并引流膽汁；分離肝門部，用無創(chuàng)血管夾夾閉肝左、中葉血管（包括動、靜脈和膽管），阻斷血流，并保持肝右、尾狀葉血流通暢，按規(guī)定時間撤夾，恢復血供，從而制成70％肝缺血再灌注損傷模型。各組SD大鼠均于缺血60min，經尾靜脈注射氫氣飽和生理鹽水，再灌注120min后統(tǒng)一取左肝組織并收集膽汁（均為I/R前后兩組）。肝組織分別用10%甲醛固定和凍存在-80℃冰箱內，HE染色后光鏡病理學觀察，觀察肝組織和肝內膽管上皮細胞的病理形態(tài)變化；采用免疫組織化學法（PV二步法）觀察肝組織中Bcl-2/Fas蛋白在肝內膽管上皮細胞的表達情況；Tunel法檢測細胞凋亡情況；分光光度法檢測丙二醛(mal0ndia1dehyde，MDA)和谷胱甘肽(glutathione， GSH)胡含量，超氧化物歧化酶(superoxidedismutase，SOD)和過氧化氫酶(catalase，CAT)的活力。將上面所有得到的數(shù)據(jù)用均數(shù)±標準差（X±s）表示，，采用方差分析對樣本均數(shù)進行比較；采用snk法對兩兩進行比較,使用SPSS17.0統(tǒng)計軟件進行統(tǒng)計學處理；統(tǒng)計檢驗P0.05被認為差異具有統(tǒng)計學意義。結果：假手術組（SO組），缺血再灌注組（IR組），缺血再灌注+生理鹽水組（IR+NS組）相比較，術后缺血再灌注+富氫水組（IR+HS組）的肝組織病理情況有所改善；膽汁中葡萄糖（Glu）的含量在IR前后升高的幅度明顯降低（IR后值均大于IR前值）；膽汁中γ-谷酰轉移酶（GGT）的活性在IR前后升高的幅度明顯明顯降低（IR后值均大于IR前值）；IR組、IR+NS組、IR+HS組動物于建模術后肝組織內MDA含量明顯升高，GSH含量降低，SOD活力和CAT活力下降;與IR+NS組相比較，IR+HS組術后肝組織內MDA含量升高較少（P0.01）.，GSH含量下降程度降低（P0.01），SOD活力和CAT活力下降程度也降低（P0.01）。免疫組化法測bcl-2基因和Fas基因的表達，IR+HS組肝內膽管上皮細胞陽性細胞數(shù)明顯比IR+NS組減少。Tunel檢測肝內膽管上皮細胞凋亡，IR+HS組肝內膽管上皮細胞凋亡明顯比IR+NS組減輕。結論:氫氣飽和生理鹽水可以減輕肝缺血再灌注后肝內膽道損傷，可以抑制肝臟缺血再灌注損傷所引發(fā)的氧化應激反應，減輕氧化損傷，可以抑制肝缺血再灌注后肝內膽管上皮細胞Bcl-2／Fas蛋白的表達，減少細胞凋亡。
[Abstract]:Objective : To study the possible mechanism of hydrogen saturated physiological saline by establishing an animal model of intra - hepatic bile duct ischemia - reperfusion injury ( IR ) in rats . The mechanism of possible mechanism of hydrogen saturated physiological saline was investigated in the following aspects : sham operation group ( SO group ) , ischemia / reperfusion group ( IR group ) , ischemia / reperfusion + physiological saline group ( IR + NS group ) , ischemia / reperfusion + hydrogen saturated physiological saline group ( IR + HS group ) .
The liver tissues were isolated from left and middle lobe vessels ( including dynamic , venous and bile duct ) , the blood flow was blocked and the blood supply was recovered after reperfusion for 120 min after reperfusion for 120 min . The pathological changes of liver and intrahepatic bile duct epithelial cells were observed .
The expression of Bcl - 2 / Fas protein in liver tissues was observed by immunohistochemical method ( PV - 2 ) .
Tunel method was used to detect the apoptosis of cells .
The activity of malondialdehyde ( MDA ) and glutathione ( GSH ) , superoxide dismutase ( SOD ) and catalase ( CAT ) were determined by spectrophotometry . All the data obtained were expressed by mean 鹵 standard deviation ( X 鹵 s ) , and the mean number of samples was compared by variance analysis .
Using the snk method , the two methods were compared , and the SPSS 17.0 statistical software was used for statistical processing ;
The results were as follows : sham operation group ( SO group ) , ischemia / reperfusion group ( IR group ) , ischemia / reperfusion + physiological saline group ( IR + NS group ) .
The content of Glu in bile was significantly decreased before and after IR ( both IR and IR values were higher than those before IR ) .
The activity of 緯 - glutamyltransferase ( GGT ) in bile was significantly lower than that before and after IR ( both IR and IR values were higher than those before IR ) .
Compared with IR + NS group , the content of MDA in the liver tissues of IR + HS group decreased significantly ( P0.01 ) . The expression of bcl - 2 gene and Fas gene was decreased in IR + HS group .

【學位授予單位】：瀘州醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R657.3

【參考文獻】

相關期刊論文 前1條

1 駱助林;湯禮軍;汪濤;羅浩;黃竹;王華;田伏洲;;氫氣生理鹽水對大鼠肝缺血再灌注后氧化應激損傷的保護作用[J];創(chuàng)傷外科雜志;2013年02期

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