發(fā)布時間：2018-05-02 10:39

  本文選題：小型豬 + 肝臟缺血再灌注��； 參考：《東北農(nóng)業(yè)大學》2017年碩士論文

【摘要】：在復雜的肝臟手術中常會出現(xiàn)缺血再灌注現(xiàn)象,可造成肝臟嚴重損傷,如何有效減輕手術造成的肝損傷是臨床上亟待解決的問題。最近幾年醫(yī)學氣體在臨床中得到了廣泛應用,經(jīng)研究表明氫氣(H_2)不僅能夠抗氧化,同時還能夠有效減輕炎癥反應,在眾多器官疾病中具有重要作用。目前對于富氫生理鹽水(HRS)的研究大多局限于大鼠、兔等小型實驗動物,對于與人同源性近的小型豬的研究極少,而將HRS應用于腹腔鏡技術建立的肝臟缺血再灌注合并肝損傷模型的研究尚未見報道。因此本研究應用HRS干預小型豬腹腔鏡微創(chuàng)肝臟手術,通過跟蹤監(jiān)測術后小型豬氧化應激和炎癥反應的變化,來深入探討HRS干預后對機體氧化應激和炎癥反應的影響。本實驗將24頭小型豬隨機分為假手術組(A組)、模型組(B組)和HRS干預組(C組)。A組僅進行氣腹并翻動肝葉,B組建立手術模型,C組建立手術模型并于再灌注前10 min、術后1 d、2 d、3 d經(jīng)門靜脈置管注射HRS(10 m L/kg)。各組分別于術前、再灌注后即刻、2h、6 h、術后1 d、2 d、3 d、5 d、7 d采集血液樣本,進行氧化應激及炎癥指標的檢測,比較組間差異。本實驗成功完成各組手術,均未發(fā)生嚴重并發(fā)癥。氧化應激檢測發(fā)現(xiàn):A組SOD、CAT、MPO、MDA在再灌注后6 h與術前比較差異顯著(0.01P0.05),之后迅速恢復,表明麻醉氣腹對機體氧化應激影響輕微且短暫;B組SOD、CAT、GSH-Px、MPO在再灌注后2 h與術前和A組比較差異極顯著(P0.01),MDA水平上升幅度高于A組,在再灌注后6 h與A比較差異顯著(0.01P0.05),說明模型對機體在再灌注早期能夠造成嚴重影響;C組MPO在再灌注后2 h與B組相比差異極顯著(P0.01),MDA在再灌注后6 h低于B組,與B組相比差異顯著(0.01P0.05),代表HRS能夠減輕模型造成的氧化應激反應,減少脂質過氧化產(chǎn)物。炎癥指標檢測發(fā)現(xiàn):A組WBC、LY、IL-6、CRP水平術后逐漸升高,在再灌注后6h與術前比較差異極顯著(P0.01);B組各項指標在再灌注后6 h、術后1 d、2 d與術前和A組比較差異極顯著(P0.01),LY在術后1 d、2 d與A組相比差異顯著(0.01P0.05),均高于A組,結果說明假手術對炎癥的影響較小并在術后很快恢復,而模型造成機體炎癥反應劇烈;C組TNF-α、IL-1β、IL-6、CRP、COR低于B組,在術后1 d、2 d與術前和B組相比差異極顯著(P0.01),IL-10在術后2 d、3 d、5 d與B組相比差異極顯著(P0.01),結果表明C組氧化應激和炎癥反應較B組輕。綜上所述得出結論:本實驗成功完成了肝門靜脈置管,將HRS應用于小型豬腹腔鏡微創(chuàng)肝臟手術模型,為HRS的臨床應用奠定了基礎,拓寬了實驗動物模型,為HRS對腹腔鏡微創(chuàng)肝臟手術的作用研究提供科學的理論依據(jù);HRS能有效提高小型豬術后抗氧化酶的活力,減輕肝臟缺血再灌注合并肝損傷引起的氧化應激反應;HRS通過抑制促炎因子,增加抗炎因子的表達,減輕術后機體的炎癥反應。
[Abstract]:Ischemia and reperfusion often occur in complicated liver surgery, which can cause serious liver injury. How to effectively alleviate the liver injury caused by surgery is a clinical problem to be solved. In recent years, medical gas has been widely used in clinic. Studies show that hydrogen gas H2) not only can resist oxidation, but also can effectively reduce inflammatory reaction, which plays an important role in many organ diseases. At present, most of the studies on HRSs are limited to small experimental animals, such as rats and rabbits, but very little to miniature pigs with close homology with human beings. However, the application of HRS to the model of liver ischemia reperfusion combined with liver injury established by laparoscopic technique has not been reported. In this study, HRS was used to intervene mini-pig laparoscopic minimally invasive liver surgery, and the effects of HRS intervention on oxidative stress and inflammatory response were studied by monitoring the changes of oxidative stress and inflammatory response in mini-pigs after operation. In this experiment, 24 miniature pigs were randomly divided into sham-operation group (group A, model group B) and HRS intervention group (group C, group C). HRS(10 mL / kg was injected into the portal vein at the end of 2 days and 3 days after operation. Blood samples were collected before operation, immediately after reperfusion for 6 hours and 1 day after operation for 2 days and 3 days and 5 days for 7 days, respectively. The indexes of oxidative stress and inflammation were detected, and the differences between the groups were compared. The operation was successfully completed in each group without serious complications. Oxidative stress test showed that there was a significant difference in MDA between before and after reperfusion at 6 h after reperfusion, and then recovered rapidly. The results showed that the effect of anesthesia pneumoperitoneum on oxidative stress was slight and the level of MDA in group B was significantly higher than that in group A at 2 h after reperfusion. There was a significant difference between group A and group A at 6 h after reperfusion, indicating that the MPO in group C was significantly lower than that in group B at 6 h after reperfusion compared with group B at 2 h after reperfusion. Compared with group B, there was a significant difference between group B and group B, indicating that HRS could attenuate the oxidative stress reaction and reduce the product of lipid peroxidation. The inflammatory index test showed that the level of IL-6 CRP increased gradually after operation in group WBCU. There were significant differences in the indexes of group B and group A at 6 h after reperfusion, 1 day after reperfusion and 1 day after reperfusion. There was a significant difference between group A and group A on the 1st day and 2nd day after reperfusion, and there was a significant difference between group A and group A on the 1st day and 2nd day after reperfusion, which was significantly higher than that in group A (P 0.01 P 0.05), and was significantly higher than that in group A (P 0.01 P 0.05) at 1 day after reperfusion, and was significantly higher than that in group A (P 0.01 P 0.05). The results showed that the effect of sham operation on inflammation was small and recovered quickly after operation, but the model caused severe inflammatory reaction in C group (TNF- 偽 IL-1 尾) and IL-6 CRPCOR was lower than that in B group. There was a significant difference in IL-10 between group B and group B on the 1st and 2nd day after operation. The results showed that the oxidative stress and inflammatory response in group C were lighter than those in group B. To sum up, this experiment successfully completed hepatic portal vein catheterization, and applied HRS to mini-pig laparoscopic minimally invasive liver surgery model, which laid the foundation for clinical application of HRS and widened the experimental animal model. To provide a scientific theoretical basis for the study of the effect of HRS on laparoscopic minimally invasive liver surgery. The oxidative stress response induced by liver ischemia / reperfusion injury was alleviated. HRS alleviated the inflammatory response after operation by inhibiting the expression of pro-inflammatory factor and increasing the expression of anti-inflammatory factor.
【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】：2017
【分類號】：R657.3;R-332

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