大鼠回肠贮袋炎的肠屏障变化研究

发布时间：2018-05-31 16:05

本文选题：回肠贮袋炎 + 肠屏障　；参考：《天津医科大学》2015年硕士论文

【摘要】：目的:建立大鼠回肠贮袋炎模型,研究大鼠回肠贮袋炎的肠屏障变化,并探讨其发生改变的机制。方法:利用显微外科技术建立大鼠回肠贮袋模型,将模型大鼠随机分为IPAA组(接受回肠贮袋手术,术后正常饮水)和DSS组(接受回肠贮袋手术,术后第30天始,自由饮用5%DSS溶液4天),并设假手术组(接受开关腹手术,术后正常饮水)作为对照。实验期间观察记录各组大鼠体重变化、便血情况、粪便评分;术后第35天,处死全部大鼠,假手术组取末端回肠组织,IPAA组及DSS组取贮袋组织及贮袋近端回肠组织,HE染色进行组织学评分;免疫组化染色检测组织MPO、occludin蛋白表达水平;留取血标本,采用比色法测定血D-乳酸浓度;实时荧光定量PCR检测组织IL-1β、IL-6、TNF-α和IL-10的m RNA转录水平。结果:各组大鼠术后体重在短期下降之后逐渐增长,术后第30-35天,DSS组体重再次持续下降,假手术组和IPAA组则继续增长;DSS组大鼠均出现血便(6/6),假手术组(0/6)和IPAA组(0/6)均无便血;DSS组粪便评分低于假手术组(P0.01)和IPAA组(P0.01),IPAA组粪便评分低于假手术组(P0.01);DSS组和IPAA组贮袋的组织学评分均高于DSS组回肠、IPAA组回肠和假手术组回肠(P0.01),且DSS组贮袋高于IPAA组贮袋(P0.01),而各组回肠的组织学评分之间差异无统计学意义(P0.05)。DSS组和IPAA组的MPO蛋白表达水平,血D-乳酸浓度,IL-1β、IL-6和TNF-α的m RNA转录水平均高于假手术组(P0.01),且DSS组均高于IPAA组(P0.01)。DSS组和IPAA组的occludin蛋白表达水平和IL-10m RNA转录水平均低于假手术组(P0.01),且DSS组均低于IPAA组(P0.01)。结论:1.成功建立大鼠回肠贮袋模型,并使用DSS成功诱导大鼠回肠贮袋炎模型。2.大鼠回肠贮袋炎模型的肠屏障结构和功能受损,受损程度和炎症程度呈正相关。3.促炎因子相对增多和抗炎因子相对减少促进肠屏障受损。4.针对促炎因子、抗炎因子进行的治疗,可能对肠屏障结构和功能的恢复有效。
[Abstract]:Aim: to establish a rat model of ileal pouch inflammation, to study the changes of intestinal barrier in rats with ileal pouch inflammation, and to explore the mechanism of the changes. Methods: rat ileal bag model was established by microsurgical technique. The model rats were randomly divided into two groups: IPAA group (receiving ileal bag operation, normal drinking water after operation) and DSS group (receiving ileal bag operation 30 days after operation). Free drinking of DSS solution for 4 days, and sham operation group (operation on switch abdomen, normal drinking water after operation) as control group. During the experiment, the changes of body weight, hematochezia and stool score were observed and recorded, and all the rats were killed on the 35th day after operation. In the sham operation group, the terminal ileum tissue of IPAA group and the DSS group were taken from the bag tissue and the proximal ileum tissue for histological grading; the expression of MPOOOOcludin protein was detected by immunohistochemical staining; the blood samples were collected. The serum Dlactic acid concentration was measured by colorimetry, and the mRNA transcription levels of IL-6TNF- 伪 and IL-10 were detected by real-time fluorescence quantitative PCR. Results: the weight of the rats in each group increased gradually after the short-term decrease, and the weight of the DSS group decreased again on the 30th to 35th day after operation. In the sham operation group and the IPAA group, the stool score in the DSS group was lower than that in the sham operation group and the IPAA group. The stool score in the DSS group was lower than that in the sham operation group (P 0.01) and the stool score in the IPAA group was lower than that in the sham operation group (P 0.01) and in the IPAA group (P 0.01 / 6). The stool score in the DSS group was lower than that in the sham operation group (P 0.01) and the stool score in the IPAA group was lower than that in the IPAA group. The histological scores of DSS group were higher than that of DSS group IPAA group and sham operation group (P 0.01), and that of DSS group was higher than that of IPAA group (P 0.01), but there was no significant difference in histological score between DSS group and IPAA group (P 0.05) and IPAA group (P < 0.05), the expression level of MPO protein in DSS group was higher than that in IPAA group, and the expression of MPO protein in DSS group was higher than that in IPAA group. The mRNA transcription levels of IL-1 尾 and TNF- 伪 in serum DLA were higher than those in sham operation group (P 0.01), and the levels of occludin protein expression and IL-10 mRNA transcription in DSS group were higher than those in IPAA group (P 0.01) and IPAA group (P 0.01), and the levels of IL-10 mRNA transcription in DSS group were all lower than those in IPAA group (P 0.01). Conclusion 1. The rat ileal bag model was successfully established and the rat ileal pouch model was successfully induced by DSS. The intestinal barrier structure and function of rat ileal pouch model were damaged, and the degree of injury was positively correlated with the degree of inflammation. The relative increase of proinflammatory factor and the decrease of anti-inflammatory factor promote intestinal barrier damage. 4. The treatment of proinflammatory factor and anti-inflammatory factor may be effective for the recovery of intestinal barrier structure and function.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R656.9

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