HRS干预小型猪腹腔镜微创肝脏手术对机体氧化应激和炎性反应的研究

发布时间：2018-05-02 10:39

本文选题：小型猪 + 肝脏缺血再灌注　；参考：《东北农业大学》2017年硕士论文

【摘要】：在复杂的肝脏手术中常会出现缺血再灌注现象,可造成肝脏严重损伤,如何有效减轻手术造成的肝损伤是临床上亟待解决的问题。最近几年医学气体在临床中得到了广泛应用,经研究表明氢气(H_2)不仅能够抗氧化,同时还能够有效减轻炎症反应,在众多器官疾病中具有重要作用。目前对于富氢生理盐水(HRS)的研究大多局限于大鼠、兔等小型实验动物,对于与人同源性近的小型猪的研究极少,而将HRS应用于腹腔镜技术建立的肝脏缺血再灌注合并肝损伤模型的研究尚未见报道。因此本研究应用HRS干预小型猪腹腔镜微创肝脏手术,通过跟踪监测术后小型猪氧化应激和炎症反应的变化,来深入探讨HRS干预后对机体氧化应激和炎症反应的影响。本实验将24头小型猪随机分为假手术组(A组)、模型组(B组)和HRS干预组(C组)。A组仅进行气腹并翻动肝叶,B组建立手术模型,C组建立手术模型并于再灌注前10 min、术后1 d、2 d、3 d经门静脉置管注射HRS(10 m L/kg)。各组分别于术前、再灌注后即刻、2h、6 h、术后1 d、2 d、3 d、5 d、7 d采集血液样本,进行氧化应激及炎症指标的检测,比较组间差异。本实验成功完成各组手术,均未发生严重并发症。氧化应激检测发现:A组SOD、CAT、MPO、MDA在再灌注后6 h与术前比较差异显著(0.01P0.05),之后迅速恢复,表明麻醉气腹对机体氧化应激影响轻微且短暂;B组SOD、CAT、GSH-Px、MPO在再灌注后2 h与术前和A组比较差异极显著(P0.01),MDA水平上升幅度高于A组,在再灌注后6 h与A比较差异显著(0.01P0.05),说明模型对机体在再灌注早期能够造成严重影响;C组MPO在再灌注后2 h与B组相比差异极显著(P0.01),MDA在再灌注后6 h低于B组,与B组相比差异显著(0.01P0.05),代表HRS能够减轻模型造成的氧化应激反应,减少脂质过氧化产物。炎症指标检测发现:A组WBC、LY、IL-6、CRP水平术后逐渐升高,在再灌注后6h与术前比较差异极显著(P0.01);B组各项指标在再灌注后6 h、术后1 d、2 d与术前和A组比较差异极显著(P0.01),LY在术后1 d、2 d与A组相比差异显著(0.01P0.05),均高于A组,结果说明假手术对炎症的影响较小并在术后很快恢复,而模型造成机体炎症反应剧烈;C组TNF-α、IL-1β、IL-6、CRP、COR低于B组,在术后1 d、2 d与术前和B组相比差异极显著(P0.01),IL-10在术后2 d、3 d、5 d与B组相比差异极显著(P0.01),结果表明C组氧化应激和炎症反应较B组轻。综上所述得出结论:本实验成功完成了肝门静脉置管,将HRS应用于小型猪腹腔镜微创肝脏手术模型,为HRS的临床应用奠定了基础,拓宽了实验动物模型,为HRS对腹腔镜微创肝脏手术的作用研究提供科学的理论依据;HRS能有效提高小型猪术后抗氧化酶的活力,减轻肝脏缺血再灌注合并肝损伤引起的氧化应激反应;HRS通过抑制促炎因子,增加抗炎因子的表达,减轻术后机体的炎症反应。
[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- 伪 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.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R657.3;R-332

【参考文献】