發(fā)布時(shí)間：2018-02-13 23:13

  本文關(guān)鍵詞： 間斷低氧預(yù)適應(yīng) 缺血再灌注損傷 Bcl-2 Bax 凋亡 肝臟　出處：《揚(yáng)州大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：1、目的 建立大鼠肝切除殘余肝臟缺血再灌注損傷模型,觀察術(shù)前間斷低氧預(yù)適應(yīng)對(duì)殘余肝臟中凋亡相關(guān)蛋白Bcl-2和Bax表達(dá)的影響。進(jìn)而探討間斷低氧預(yù)適應(yīng)對(duì)肝切除合并缺血再灌注損傷肝臟的保護(hù)機(jī)制,為其在肝臟外科臨床應(yīng)用尋找理論依據(jù)。 2、方法 健康SD大鼠20只,用于大鼠肝臟解剖和肝切除缺血再灌注損傷模型的練習(xí),計(jì)算模型術(shù)后一周生存率。能夠建立穩(wěn)定的模型后,另取SD大鼠54只,隨機(jī)分為肝切除組(PH組)、肝切除殘余肝臟缺血再灌注組(IR組)和術(shù)前間斷低氧預(yù)適應(yīng)組(IHP組)。分別結(jié)扎PH組大鼠肝臟左葉和中葉根部后將其切除,切除部分濕重約占整個(gè)肝臟濕重的70%。殘余肝臟缺血再灌注組(IR組),即在夾閉肝十二指腸韌帶、阻斷入肝血流情況下,按照PH組的方法切除肝臟的左葉和中葉。肝門(mén)阻斷20min后開(kāi)放血流,殘余肝臟發(fā)生了缺血再灌注過(guò)程。間斷低氧預(yù)適應(yīng)組(IHP組),術(shù)前一周將大鼠置于氧氣體積分?jǐn)?shù)10%的低氧環(huán)境中接受低氧刺激,每天1h。一周后在肝門(mén)阻斷下行肝切除術(shù)(同IR組)。各組分別于術(shù)后12、24和48h取下腔靜脈血和殘余肝組織進(jìn)行檢測(cè)。用全自動(dòng)生化分析儀檢測(cè)血清谷丙轉(zhuǎn)氨酶(ALT)、谷草轉(zhuǎn)氨酶(AST)含量,間接反映肝臟受損程度。采用免疫組織化學(xué)方法檢測(cè)殘余肝組織凋亡相關(guān)蛋白Bcl-2和Bax表達(dá)情況,以積分光密度來(lái)反映其表達(dá)強(qiáng)度。 3、結(jié)果 1.肝大部切除合并殘余肝臟缺血再灌注損傷模型術(shù)后七天生存率為93.3%。麻醉復(fù)蘇后大鼠健康存活,未發(fā)現(xiàn)明顯手術(shù)并發(fā)癥。 2.術(shù)后肝功能在術(shù)后每個(gè)時(shí)間點(diǎn),IR組和IHP組大鼠血清ALT和AST水平均高于PH組,但I(xiàn)HP組明顯低于IR組,差異均具有統(tǒng)計(jì)意義(P0.05)。 3.術(shù)后殘余肝臟組織中Bcl-2和Bax表達(dá)’術(shù)后每個(gè)時(shí)間點(diǎn)各組大鼠肝臟中Bcl-2蛋白表達(dá)的積分光密度相比,IHP組均顯著高于PH組和IR組(差異有統(tǒng)計(jì)學(xué)意義,P0.05)。雖然PH組在術(shù)后每個(gè)時(shí)間點(diǎn)也均高于IR組,但僅在術(shù)后48小時(shí)有統(tǒng)計(jì)學(xué)差異(P0.05),術(shù)后12小時(shí)和24小時(shí)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。Bax蛋白表達(dá)的積分光密度與Bcl-2相反,IHP組在各時(shí)間點(diǎn)均低于IR組,但高于PH組。差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。 4、結(jié)論 通過(guò)臨時(shí)夾閉肝十二指腸韌帶,可建立穩(wěn)定、可靠的肝切除殘余肝臟缺血再灌注模型(肝再生合并缺血再灌注損傷模型)。間斷低氧預(yù)適應(yīng)對(duì)殘余肝臟缺血再灌注損傷具有保護(hù)作用,其途徑可能是通過(guò)促進(jìn)抗凋亡蛋白Bcl-2表達(dá)和抑制促凋亡蛋白Bax表達(dá),來(lái)減少肝細(xì)胞凋亡。
[Abstract]:1. Purpose. A rat model of hepatic ischemia-reperfusion injury after hepatectomy was established. To observe the effect of intermittent hypoxia preconditioning on the expression of apoptosis-related protein Bcl-2 and Bax in residual liver, and to explore the protective mechanism of intermittent hypoxia preconditioning on liver injury after hepatectomy and ischemia reperfusion injury. To find a theoretical basis for its clinical application in liver surgery. 2, method. Twenty healthy Sprague-Dawley (SD) rats were used for the exercise of liver anatomy and hepatectomy ischemia-reperfusion injury model, and the one-week survival rate of the model was calculated. After establishing a stable model, another 54 SD rats were taken out. The rats were randomly divided into hepatectomy group (PH group), hepatectomy residual liver ischemia reperfusion group (IR group) and intermittent hypoxia preconditioning group (IHP group). Zuo Ye of liver and middle lobe root of PH group were ligated respectively. The partial wet weight of the liver was about 70% of the total wet weight of the liver. In the residual liver ischemia-reperfusion group, the IR group was in the condition of clamping the ligaments of the liver and duodenum and blocking the blood flow into the liver. Zuo Ye and middle lobe of the liver were resected according to PH group. The hepatic portal was blocked for 20 minutes and the blood flow was opened. The rats in the intermittent hypoxic preconditioning group were exposed to hypoxic stimulation in hypoxic environment with oxygen volume fraction 10% one week before operation. After 1 hour a day, the hepatic resection was performed at hepatic hilus occlusion (IR group). The blood samples of inferior vena cava and residual liver tissue were taken at 1224 and 48 hours after operation. The serum alanine aminotransferase (alt) and alanine aminotransferase (alt) were measured by automatic biochemical analyzer. Immunohistochemical method was used to detect the expression of apoptosis-related protein Bcl-2 and Bax in residual liver tissue, and the intensity of expression was reflected by integral optical density. 3, result. 1. The survival rate of the model of hepatic ischemia reperfusion injury after subtotal hepatectomy was 93.30.The survival rate of rats after anesthesia and resuscitation was healthy and there were no obvious operative complications. 2. The levels of serum ALT and AST in IR group and IHP group were higher than those in PH group at each time point after operation, but those in IHP group were significantly lower than those in IR group. The difference was statistically significant (P 0.05). 3. Expression of Bcl-2 and Bax in residual liver tissue after operation: the integral optical density of Bcl-2 protein expression in the liver of each group was significantly higher than that of PH group and IR group at each time point after operation (P < 0.05), although the expression of Bcl-2 protein in PH group was significantly higher than that in PH group and IR group (P < 0.05). Each time point after operation was also higher than that in IR group. However, there was a significant difference only at 48 hours after operation (P 0.05). There was no significant difference between 12 hours and 24 hours after operation. The integral optical density of P0.05U. Bax protein expression in Bcl-2 group was lower than that in IR group at all time points, but higher than that in PH group. The difference was statistically significant (P 0.05). 4. Conclusion. Stability can be established by temporarily clipping the hepatoduodenal ligament. A reliable model of residual liver ischemia-reperfusion injury after hepatectomy (liver regeneration combined with ischemia-reperfusion injury). Intermittent hypoxic preconditioning has protective effect on residual liver ischemia-reperfusion injury. The pathway may be to reduce hepatocyte apoptosis by promoting the expression of anti-apoptotic protein Bcl-2 and inhibiting the expression of pro-apoptotic protein Bax.
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R657.3

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