鱼油对脓毒症幼鼠肠道菌群、细菌易位及炎症因子TNF-a的影响

发布时间：2018-04-04 09:23

本文选题：鱼油　切入点：脓毒症　出处：《南昌大学》2017年硕士论文

【摘要】：目的:研究脓毒症幼鼠肠道菌群紊乱、易位和血清炎症因子TNF-α的变化,以及鱼油干预后对上述情况的影响。方法:1、实验分组及喂养方式:选取24只健康SD幼鼠(4周龄),根据完全随机分组原则分为三组:正常对照组,模型组及鱼油治疗组(每组各8只)。正常组为正常状态每日饲喂基础日粮,模型组饲喂每日基础日粮,鱼油治疗组则是基础日粮加鱼油灌胃。按上述方式分别喂养1周。第8天时模型组、鱼油治疗组幼鼠皆制作脓毒症模型,并依照5mg/kg的剂量经腹腔注射制剂——大肠杆菌脂多糖(LPS)的方法造模,三组继续如上述喂养方式喂养3d。2、标本采集:制模第三天下午2时开胸收集实验所需各组织,心脏取血用于ELISA实验测定血清TNF-α,采集并准确称量肝、脾、肾、肠系膜淋巴结、结肠内容物各1g,在距离屈氏韧带8厘米处迅速切除约10厘米回肠段置于福尔马林中浸泡以用于病理学检查。3、观察指标:观察各组幼鼠肠道菌群(双歧杆菌、乳酸杆菌、大肠杆菌、葡萄球菌)及细菌易位率的变化及比较;ELISA实验测定各组幼鼠血清炎症因子TNF-α数值变化;采用苏木精—伊红染色法(HE染色),显微镜下取每组(各组随机2只)幼鼠回肠组织观察病理学改变。结果:1、一般表现:模型组制模后出现病态表现:精神差,活动、食纳减少,寒战竖毛,呼吸增快,口唇发绀。剖腹时可见少数幼鼠存在腹腔积液,部分肠管肿胀、发黑。鱼油治疗组与模型组对比,各病态表现均有所减轻,正常对照组无上述病态表现。2、脓毒症幼鼠存在肠道菌群紊乱状态,在细菌具体数量上表现为肠道双歧杆菌及肠道乳酸杆菌明显下降(P0.01),肠道大肠杆菌及葡萄球菌则明显升高(P0.01),鱼油对脓毒症幼鼠菌群紊乱具有一定纠正作用(P0.01)。它增加了上述肠道双歧、乳酸杆菌的数目(P0.01),降低了大肠杆菌、葡萄球菌的数目(P0.01),减少了肠道细菌易位的发生。3、脓毒症幼鼠血清炎症因子TNF-α明显升高(P0.01),鱼油能显著减低TNF-α数值(P0.01)。4、脓毒症幼鼠存在肠道粘膜组织学损伤,鱼油可改善脓毒症幼鼠回肠黏膜损伤和回肠组织病理变化。结论:脓毒症幼鼠肠道菌群存在紊乱及易位,伴肠道粘膜损伤,血清炎症因子TNF-α明显升高;鱼油干预可促进肠道双歧、乳酸杆菌的生长使数量增加,抑制大肠杆菌及葡萄球菌而使数量降低;可减少细菌易位的发生;对血清TNF-α的释放具有一定抑制作用,可减轻组织和器官的一系列损伤;改善了脓毒症幼鼠回肠黏膜损伤和回肠组织病理变化,使肠道免疫屏障保持完整,提高肠道黏膜修复力。
[Abstract]:Aim: to study the changes of intestinal flora disturbance, translocation and serum inflammatory factor TNF- 伪 in septic young rats.Methods: 24 healthy SD rats were randomly divided into three groups: normal control group, model group and fish oil treatment group (8 rats in each group).The normal group was fed the basic diet daily, the model group was fed the daily basic diet, and the fish oil treatment group was fed with the basic diet plus fish oil.The animals were fed for 1 week according to the above method.On the 8th day, the model group, fish oil treatment group all made sepsis model, and according to the dose of 5mg/kg through intraperitoneal injection preparation-Escherichia coli lipopolysaccharide lipopolysaccharide (LPS), the model was made.The three groups continued to feed 3d.2as mentioned above. Specimen collection: the tissues needed for the open-chest collection experiment were collected at 2: 00 p.m. on the third day of model making. Blood was taken from the heart to determine serum TNF- 伪 by ELISA test. The liver, spleen, kidney and mesenteric lymph nodes were collected and weighed accurately, and the liver, spleen, kidney and mesenteric lymph nodes were measured accurately.Colon contents, 1 g each, were quickly removed about 10 cm from the tricuspid ligament and immersed in formalin for pathological examination .3.The indicators: the intestinal flora (Bifidobacterium, Lactobacillus) was observed in each group.Comparison of the changes of Escherichia coli, Staphylococcus) and bacterial translocation rate in young rats serum TNF- 伪 was determined by Elisa.Hematoxylin-eosin staining was used to observe the pathological changes in ileum of each group (2 randomly in each group).Results: in the model group, there were pathological manifestations after modeling: mental retardation, activity, decrease of food intake, shivering hair, rapid breathing, cyanosis of lips.During laparotomy, a few young rats were found to have abdominal effusion, swelling and blackening of the intestine.Compared with the model group, the pathological manifestations of fish oil treatment group were alleviated, and the normal control group did not have the pathological manifestation. 2. The septic young rats had intestinal flora disorder state.The specific number of bacteria showed that intestinal bifidobacterium and intestinal Lactobacillus decreased significantly (P 0.01), intestinal Escherichia coli and Staphylococcus significantly increased P 0.01, fish oil had a certain corrective effect on sepsis young rat flora disorder (P0.01).It increases the number of Lactobacillus bifidus, P0. 01, and reduces the number of Escherichia coli.The number of staphylococci decreased the occurrence of intestinal bacterial translocation. The inflammatory factor TNF- 伪 in serum of septic young rats was significantly increased. Fish oil could significantly reduce the value of TNF- 伪, P0.01. 4. There were intestinal mucosal tissue damage in sepsis young rats.Fish oil can improve ileal mucosal injury and ileal histopathological changes in septic young rats.Conclusion: the intestinal flora of sepsis young rats has disorder and translocation, accompanied by intestinal mucosal injury, serum inflammatory factor TNF- 伪 is obviously increased, fish oil intervention can promote intestinal bifidus, Lactobacillus growth can increase the number.Inhibition of Escherichia coli and Staphylococcus can reduce the number of bacteria translocation can reduce the release of serum TNF- 伪 can reduce a series of tissue and organ damage.It improved the ileal mucosal injury and the pathological changes of ileum in septic young rats, kept the intestinal immune barrier intact and improved the repair ability of intestinal mucosa.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R720.597

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