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

【摘要】：目的：本實(shí)驗(yàn)通過(guò)建立大鼠肝內(nèi)膽管缺血再灌注損傷(IR)動(dòng)物模型，探討靜脈注射氫氣飽和生理鹽水是否對(duì)大鼠肝內(nèi)膽管缺血再灌注損傷具有保護(hù)作用，觀察其病理學(xué)改變，并從膽管上皮細(xì)胞Bcl-2/Fas基因表達(dá)、凋亡等方面探討氫氣飽和生理鹽水可能的作用機(jī)制。方法：將32只雄性SD大鼠隨機(jī)分成4組：假手術(shù)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(jìn)入腹腔，等待麻醉藥物效果顯效后，用碘伏對(duì)大鼠的腹部進(jìn)行消毒，然后沿著大鼠的腹正中線打開(kāi)腹部，分離至膽總管及十二指腸，并于二者匯合處經(jīng)十二指腸外用硬膜外導(dǎo)管逆行置管于膽總管內(nèi)，固定并引流膽汁；分離肝門(mén)部，用無(wú)創(chuàng)血管夾夾閉肝左、中葉血管（包括動(dòng)、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(shí)間撤夾，恢復(fù)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(jīng)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(nèi)，HE染色后光鏡病理學(xué)觀察，觀察肝組織和肝內(nèi)膽管上皮細(xì)胞的病理形態(tài)變化；采用免疫組織化學(xué)法（PV二步法）觀察肝組織中Bcl-2/Fas蛋白在肝內(nèi)膽管上皮細(xì)胞的表達(dá)情況；Tunel法檢測(cè)細(xì)胞凋亡情況；分光光度法檢測(cè)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(guò)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(biāo)準(zhǔn)差（X±s）表示，，采用方差分析對(duì)樣本均數(shù)進(jìn)行比較；采用snk法對(duì)兩兩進(jìn)行比較,使用SPSS17.0統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)學(xué)處理；統(tǒng)計(jì)檢驗(yàn)P0.05被認(rèn)為差異具有統(tǒng)計(jì)學(xué)意義。結(jié)果：假手術(shù)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(shù)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(zhuǎn)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(dòng)物于建模術(shù)后肝組織內(nèi)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(shù)后肝組織內(nèi)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(cè)bcl-2基因和Fas基因的表達(dá)，IR+HS組肝內(nèi)膽管上皮細(xì)胞陽(yáng)性細(xì)胞數(shù)明顯比IR+NS組減少。Tunel檢測(cè)肝內(nèi)膽管上皮細(xì)胞凋亡，IR+HS組肝內(nèi)膽管上皮細(xì)胞凋亡明顯比IR+NS組減輕。結(jié)論:氫氣飽和生理鹽水可以減輕肝缺血再灌注后肝內(nèi)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(yīng)激反應(yīng)，減輕氧化損傷，可以抑制肝缺血再灌注后肝內(nèi)膽管上皮細(xì)胞Bcl-2／Fas蛋白的表達(dá)，減少細(xì)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(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R657.3

【參考文獻(xiàn)】

相關(guān)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(duì)大鼠肝缺血再灌注后氧化應(yīng)激損傷的保護(hù)作用[J];創(chuàng)傷外科雜志;2013年02期

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