前列腺素E1对大鼠小肠缺血再灌注损伤保护作用的实验研究

发布时间：2018-06-24 08:41

本文选题：大鼠 + 前列腺素E1　；参考：《昆明医学院》2010年硕士论文

【摘要】： 目的：通过应用前列腺素E1对大鼠小肠缺血／再灌注损伤影响的研究,从肠粘膜的损伤程度,促炎因子和抗炎因子的释放,以及细菌移位等不同方面探讨前列腺素E1对大鼠小肠缺血再灌注损伤的是否具有保护作用,从而为临床防治小肠缺血再灌注损伤提供一个新的策略。方法：SPF级健康成年雄性SD大鼠80只,体重280g-320g,随机分为假手术(Sham group, S)组、缺血／再灌注损伤(ischemia/reperfusion injury, IRI)组、缺血预处理组(ischemia preconditioning, IPC)和前列腺E1(Prostaglandin E1,PGE1)组(n=20),每组根据再灌注后采集标本的时间分为两个亚组(n=10),实验前12h使大鼠禁食,自由饮水,并于实验前2h行含绿色荧光素蛋白大肠杆菌(Ecdi DH5α)2ml灌胃。 ①S组：麻醉后取腹正中切口,长约3cm,分离肠系膜上动脉(SMA)后关腹； ②IRI组：同S组分离SMA后,用无创微动脉夹夹闭SMA后关腹,45min后取出动脉夹,分别于再灌注2h、4h时采集标本； ③IPC组：同S组分离SMA后,给予SMA 3个循环的预处理(缺血2min后再灌注2min为一个循环)后关腹,余同IRI组； ④PGE1组：同S组分离SMA后,用无创微动脉夹夹闭SMA后关腹,缺血45min后,于再灌注前5min经尾静脉按20ug／kg注入PGE1,余同IRI组。 ELISA法检测血清中IL-1β,IL-6,IL-10和TNF-α的含量,同时取回肠壁组织长1cin行肠粘膜病理学检测并行HE染色,取肝脏组织、脾脏组织及门静脉血行细菌移位检测。结果： 1、小肠病理变化：S组大鼠肠绒毛完整,排列整齐,镜下无中性粒细胞浸润。IRI组大鼠在再灌注2h和4h后小肠绒毛上皮顶端与固有层分离并见固有层毛细血管暴露、出血和溃疡,粘膜绒毛坏死,脱落,镜下见大量中性粒细胞浸润。与IRI组相比,IPC组和PGE1组在各时间点肠粘膜绒毛损伤减轻,坏死,脱落少见,镜下中性粒细胞浸润轻。IPC组和PGE1组损伤评分明显低于IRI组(P0.05),IPC组和PGE1组相比,差异无统计学意义(P0.05)。 2、细胞因子改变：IRI组、IPC组和PGE1组再灌注2h、4h,大鼠动脉血清中炎性细胞因子IL-1β,IL-6,TNF-α含量明显高于S组(P0.05)；IPC组和PGE1组IL-1p,IL-6,TNF-a则低于IRI组(P0.05)；IPC组和PGE1组相比,差异无统计学意义(P0.05)。大鼠动脉血清中抗炎因子IL-10含量在IRI组、IPC组和PGE1组各时间点明显低于S组(氏0.05)；IPC组和PGE1组各时间点高于IRI组(P0.05)；IPC组和PGE1相比,差异无统计学意义(P0.05)。 3、细菌移位改变：再灌注2h、4h,S组大鼠脾脏、肝脏及门静脉血未检出移位细菌生长；IPC组和PGE1组明显低于IRI组(P0.05),IPC组和PGE1组相比,差异无统计学意义(P0.05)。结论： PGE1和IPC能减轻大鼠小肠缺血再灌注损伤后小肠粘膜绒毛的坏死程度：PGE1和IPC能抑制大鼠小肠缺血再灌注损伤后促炎细胞因子(IL-1p,IL-6,TNF-α)的释放和促进抗炎细胞因子IL-10的表达：PGE1和IPC能减少大鼠小肠缺血再灌注损伤后的细菌移位。
[Abstract]:Objective: to study the effects of prostaglandin E _ 1 on intestinal ischemia / reperfusion injury in rats, and to investigate the extent of intestinal mucosal injury, the release of proinflammatory and anti-inflammatory factors, and the effects of prostaglandin E _ 1 on intestinal ischemia / reperfusion injury in rats. To explore whether prostaglandin E1 has protective effect on intestinal ischemia-reperfusion injury in rats from different aspects such as bacterial translocation, so as to provide a new strategy for clinical prevention and treatment of intestinal ischemia-reperfusion injury. Methods 80 healthy male Sprague-Dawley rats of SPF grade, weighing 280g-320g, were randomly divided into sham group (S) group and ischemia/reperfusion injury-injury (IRI) group. Ischemic preconditioning group (ischemia preconditioning, IPC) and prostaglandin E1 (PGE1) group were divided into two subgroups according to the time of reperfusion. The rats were fasting and drinking freely 12 hours before the experiment. In group 1, the median incision was taken after anesthesia, about 3 cm long, and the superior mesenteric artery (SMA) was separated and then closed. 2IRI group: after SMA was separated from the same S group, the artery clamp was removed 45 minutes after the closure of the SMA, and the specimens were collected at 4 h after 2 h reperfusion, while in the same S group, the SMA was separated from the same S group. Three cycles of preconditioning (2min after ischemia and reperfusion as a circulation) were given, followed by closure of abdomen, rest in IRI group, 4PGE1 group: after isolation of SMA from group S, closure of abdomen with non-invasive microartery clamping, and after ischemia of 45min. Before reperfusion, 5min was injected into PGE1 via caudal vein according to 20ug/kg. Elisa was used to detect the contents of IL-1 尾, IL-6, IL-10 and TNF- 伪 in serum, and the long 1cin of intestinal wall was collected for intestinal mucosal pathological examination and HE staining, and liver tissue was obtained. Bacterial translocation was detected in splenic tissue and portal vein blood. Results: 1. The intestinal villi were intact and arranged neatly in the group of small intestine pathological changes. No neutrophil infiltration. IRI rats were separated from the lamina propria at 2 and 4 hours after reperfusion and showed capillary exposure hemorrhage and ulcer mucosal villi necrosis and neutrophils infiltration under microscope. Compared with IRI group, IPC group and PGE1 group had less injury, necrosis and shedding of intestinal mucosal villi at different time points. The injury scores of IPC group and PGE1 group were significantly lower than those of IRI group (P0.05) and PGE1 group (P 0.05), and the injury scores of IPC group and PGE1 group were significantly lower than those of IRI group (P0.05). There was no significant difference (P0.05). 2. The changes of cytokines in IPC group and PGE1 group were significantly higher than those in group S (P0.05), and the levels of IL-1pIL-6TNF-a in arterial serum of rats were significantly higher than those of IRI group (P0.05), and the level of IL-1pIL-6TNF-a in arterial serum of rats was significantly higher than that of IRI group (P0.05). There was no significant difference between IPC group and PGE1 group (P0.05). The levels of anti-inflammatory factor IL-10 in serum of rats in IRI group were significantly lower than those in group S (0.05) and PGE1 group (P 0.05), and were significantly higher than those in IRI group (P0.05) and IPC group and PGE1 group at each time point. There was no significant difference between the two groups (P0.05). 3. The change of bacterial translocation: the spleen, liver and portal vein blood were not detected in the spleen, liver and portal vein blood of rats in the group of 2 h reperfusion (P0.05) compared with the group of IRI (P0.05) and the group of PGE1 (PGE1), the difference of bacterial translocation between IPC group and PGE1 group was significantly lower than that in group IPC and PGE1. The difference was not statistically significant (P0.05). Conclusion: PGE1 and IPC can reduce the degree of necrosis of intestinal villi after small intestinal ischemia reperfusion injury in rats. PGE1 and IPC can inhibit the release of pro-inflammatory cytokine (IL-1pmIL-6TNF- 伪) after small intestinal ischemia-reperfusion injury in rats. And promote the expression of anti-inflammatory cytokine IL-10: PGE1 and IPC can reduce the bacterial translocation after small intestinal ischemia-reperfusion injury in rats.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

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