本文選題：遠程缺血預(yù)處理　切入點：缺血再灌注損傷　出處：《蘭州大學(xué)》2010年碩士論文

【摘要】： 目的采用兔單肺原位缺血再灌注損傷模型,通過對雙下肢遠程缺血預(yù)處理后兔單肺原位缺血再灌注損傷引起的肺組織病理學(xué)改變、MDA、SOD、細胞凋亡指數(shù)(AI) Bcl-2/Bax及血氣分析的變化,探討雙下肢遠程缺血預(yù)處理對兔單肺原位缺血再灌注損傷細胞凋亡的影響及可能機制。 方法清潔級日本大耳白兔18只,體重2.0-2.5kg,隨機分為3組(n=6)：假手術(shù)對照組(S組)、肺缺血再灌注組(IR組)、肢體遠程缺血預(yù)處理組(R組)。參照Sekido介紹的方法(略加改造)復(fù)制兔肺缺血再灌注模型：于胸骨旁左側(cè)肋軟骨與肋骨交界處向頭側(cè)剪斷第8-6肋骨,開胸暴露左肺門并留置阻斷帶,在呼氣末阻斷左肺門60min為缺血期,隨后開放阻斷帶恢復(fù)左肺供血和通氣為再灌注期。阻斷左肺門前經(jīng)耳緣靜脈給予低分子肝素1 mg·kg-1抗凝。遠程缺血預(yù)處理組(R組)在開胸左肺門放置阻斷帶穩(wěn)定30min后用阻斷帶在兔雙下肢根部同時阻斷下肢血流造成雙下肢缺血10min,以多普勒血流探測儀檢測不到股動脈搏動為準(zhǔn),然后松開阻斷帶再灌10min,重復(fù)三次。30min后結(jié)扎左肺門60min,然后恢復(fù)灌注180min。缺血再灌注組(IR組)在左肺門放置阻斷帶后曠置90min,在呼氣末阻斷左肺門60min制造肺缺血再灌注模型為缺血期,以左肺不張為準(zhǔn),60min后松開結(jié)扎線,恢復(fù)灌注1 80min,只在雙下肢松繞阻斷帶但不行雙下肢缺血。假手術(shù)組(S組)只在左肺門放置阻斷帶但不結(jié)扎肺門及只在雙下肢松繞阻斷帶但不行雙下肢缺血。分別在遠程缺血預(yù)處理前,再灌注15min、30min、60min、120min及180min時采動脈血測血氣并觀察有創(chuàng)動脈血壓的變化。各組分別于再灌注180min放血處死家兔留取標(biāo)本待測肺組織MDA含量、SOD活性、HE染色下觀察肺損傷定量評價指標(biāo)(IQA)的變化以及TUNEL法檢測肺組織凋亡指數(shù)(AI)、Westernbloting法半定量Bcl-2、Bax蛋白的表達。實驗過程中監(jiān)測直腸溫度并用電熱毯維持直腸溫度在36-38℃。 結(jié)果①PaO2, A-aDO2和RI:三組的基礎(chǔ)A-aDO2、PaO2及RI無統(tǒng)計學(xué)差異(P＞0.05),隨著再灌注時間的延長,IR組的PO2逐漸降低,尤以再灌注60min內(nèi)明顯(P0.01)。R組和S組的PaO2均高于IR組(P0.01)。IR組的A-aDO2和RI逐漸增加,尤其在再灌注60min時明顯高于R組和S組(P0.01)。再灌注后IR組的A-aDO2和RI明顯高于R組和S組(P0.01)。②IQA:IR組的IQA明顯高于R組和S組(P0.01)。③MDA含量和SOD活性：IR組的MDA含量明顯高于S組和R組(P0.01),而IR組的SOD活性明顯低于S組和R組(P0.01)。④AI和Bcl-2/Bax:與S組比較,IR組的AI和Bax蛋白含量明顯增加(P0.01),但Bcl-2明顯降低(P0.01),并且Bcl-2/Bax降低(P0.01)。與IR組比較,R組的AI和Bax明顯降低(P0.01),而Bcl-2和Bcl-2/Bax明顯增加(P0.01)。R組和S組之間比較無統(tǒng)計學(xué)意義(P0.05)。 結(jié)論雙下肢遠程缺血預(yù)處理對兔肺缺血再灌注損傷具有保護作用,其保護作用主要是通過抑制氧化損傷,上調(diào)Bcl-2/Bax的比值,從而減少肺組織細胞凋亡的發(fā)生。
[Abstract]:Objective to study the changes of Bcl-2/Bax and blood gas in lung histopathological changes induced by in situ ischemia-reperfusion injury in rabbits with single lung ischemia reperfusion injury after long leg ischemia preconditioning. To investigate the effect of remote ischemic preconditioning on apoptosis of rabbit single lung ischemia reperfusion injury and its possible mechanism. Methods Eighteen Japanese white rabbits of clean grade were used. Weight 2.0-2.5kg, randomly divided into three groups: sham operation control group S group, lung ischemia reperfusion group IR group, limb remote ischemic preconditioning group R group. According to the method introduced by Sekido (slightly modified), the rabbit lung ischemia-reperfusion model: in chest. Cut the 8-6 ribs at the junction of the left costal cartilage and the ribs at the left side of the bone to the head. Open chest exposed the left pulmonary hilum and retained the blocking band. The left pulmonary hilus 60min was blocked at the end of the breath for ischemic period. The left pulmonary portal vein was blocked with low molecular weight heparin (1 mg kg-1) for anticoagulation.The remote ischemic preconditioning group (group R) placed a blocking zone to stabilize 30min in the open left pulmonary hilum. The femoral artery pulsation could not be detected by Doppler blood flow detector after the occlusion band was used to block the lower extremity blood flow at the same time in both lower extremities of rabbits for 10 min, and the femoral artery pulsation could not be detected by Doppler blood flow detector. The left pulmonary hilum was ligated for 60 minutes after repeated for three times for 30 minutes, and then returned to perfusion for 180 minutes. In the IR group, the left pulmonary hilus was placed in the left pulmonary hilus for 90 minutes. The model of pulmonary ischemia reperfusion was established by occlusion of the left pulmonary hilus 60min at the end of expiratory period, and the model of pulmonary ischemia reperfusion was established at the end of expiratory period. The ligation line was loosened 60 minutes after the left atelectasis. Recovery perfusion for 180 min, only in both lower extremities loose occlusion zone but not lower extremity ischemia. Sham operation group (S group) only placed in the left pulmonary hilus without ligation of the pulmonary hilus, and only in the lower extremities around the blocking zone but not in both lower extremities ischemia. Before remote ischemic preconditioning, The arterial blood gas was measured and the changes of blood pressure of invasive arteries were observed during 15 minutes of reperfusion for 30 minutes and 60 minutes for 120 minutes and for 180min. The quantitative evaluation of lung injury was observed in each group of rabbits who were sacrificed by reperfusion 180min for the determination of MDA content and the activity of MDA and HE staining. The changes of IQA) and the expression of Bax protein in lung tissue were detected by TUNEL. The rectal temperature was monitored and the rectal temperature was maintained at 36-38 鈩，

本文編號：1689528