肥胖对Helicobacter suis感染后胃黏膜相关淋巴组织（MALT）形成的影响

发布时间：2018-04-08 09:26

本文选题：肥胖　切入点：H.suis　出处：《青岛大学》2017年硕士论文

【摘要】：目的通过对Helicobacter suis(H.suis)感染肥胖小鼠的研究,探讨高脂饮食诱导的肥胖对H.suis感染后小鼠胃黏膜相关淋巴样组织(mucosa associated lymphoid tissue,MALT)形成的影响及其可能的机制,进而有望为肥胖的促瘤机制提供新的研究方向。方法1.H.suis感染小鼠模型的建立:将40只雌性C57BL/6小鼠随机分为模型组和对照组,模型组每只小鼠给予0.5 m L经PCR检测确定H.suis存在的猪胃黏膜匀浆液灌胃,对照组则给予等量磷酸盐缓冲液(phosphate buffer saline,PBS)灌胃。分别于灌胃1个月及3个月后处死两组中的半数小鼠,取其胃黏膜组织,采用PCR方法检测H.suis在小鼠胃内的定植情况,HE染色方法检测胃黏膜淋巴滤泡形成的情况。2.肥胖对H.suis感染后胃MALT形成及相关炎症因子表达的影响:建模成功后,新购40只雌性C57BL/6小鼠随机分为4组,每组10只,即正常对照组(NC组)、高脂饮食组(HFD组)、H.suis感染组(HS组)和H.suis感染+高脂饮食组(HS+HFD组),将已经感染H.suis小鼠的胃黏膜匀浆液(0.2ml/只)灌胃于HS组与HS+HFD组。NC组与HS组持续给予普通饮食,HFD组与HS+HFD组自感染后第3周开始给予高脂饮食;差异性饮食24周后,处死各组小鼠取其胃组织,采用HE染色方法检测各组小鼠胃黏膜淋巴滤泡的数量与大小;采用Real-time PCR方法检测各组小鼠胃黏膜瘦素(leptin)及其受体Ob-R、炎症因子IFN-γ与趋化因子CXCL13、CCL2的表达量。结果1.灌胃所用猪胃黏膜匀浆液中存在H.suis;H.suis可以在小鼠胃内定植,1个月及3个月后均检测到该菌;H.suis感染可诱导小鼠胃黏膜滤泡的形成,并且感染3个月较感染1个月的淋巴滤泡明显增大。2.与HS组相比,HS+HFD组小鼠胃黏膜淋巴滤泡的数量与大小明显增加,胃黏膜leptin及其受体Ob-R,IFN-γ、CXCL13、CCL2的m RNA表达量亦明显增加。结论1.成功建立了稳定可靠的H.suis感染小鼠模型;2.H.suis可以诱导出胃黏膜淋巴滤泡的形成;3.高脂饮食诱导的肥胖可能促进了leptin→IFNγ→CXCL13链的表达,进而促进H.suis感染后胃黏膜相关淋巴组织的形成。
[Abstract]:Objective to investigate the effect of obesity induced by high fat diet on the formation of mucosa associated lymphoid tissue (malt) in mice infected with Helicobacter suis (H. suis) and its possible mechanism.It is expected to provide a new research direction for the mechanism of obesity.Methods: 40 female C57BL/6 mice were randomly divided into model group and control group. Each mouse in the model group was given 0.5 mL PCR assay to determine the presence of H.suis in the gastric mucosa homogenate.The control group was given the same amount of phosphate buffer solution (Phosphate buffer saline).After 1 month and 3 months of gastric perfusion, half of the mice in the two groups were killed, and their gastric mucosal tissues were taken. The colonization of H.suis in the stomach of mice was detected by PCR method. The formation of lymphatic follicles in gastric mucosa was detected by HE staining.Effects of obesity on gastric MALT formation and expression of related inflammatory factors after H.suis infection: after successful modeling, 40 newly purchased female C57BL/6 mice were randomly divided into 4 groups, 10 in each group.The normal control group (NC group), the high-fat diet group (HFD group) and the high-fat diet group (HS group) and the H.suis infected high-fat diet group (HS HFD group) were perfused with 0.2 ml / mouse gastric mucosal homogenate of infected H.suis mice. The gastric mucosa homogenate of the infected mice was given intragastric to HS group and HS HFD group. NC group and HS group were maintained in HS group and HS group.The high fat diet was given to the general diet group and HS HFD group from the 3rd week after infection.After 24 weeks of different diet, the gastric tissues of each group were killed, and the number and size of lymphatic follicles in gastric mucosa of each group were detected by HE staining.The expression of leptin and its receptor Ob-R-, IFN- 纬 and CXCL13 CCL2 in gastric mucosa of mice in each group were detected by Real-time PCR method.Result 1.H. suis could be colonized in the gastric mucosa homogenate of pigs. After 1 and 3 months, it was found that H. suis infection could induce the formation of gastric follicles in mice.The number of lymphatic follicles in 3 months of infection was significantly larger than that in 1 month of infection.Compared with HS group, the number and size of lymphoid follicles in HS HFD group were significantly increased, and the expression of m RNA in gastric mucosal leptin and its receptor Ob-R- IFN- 纬 -CXCL13CCL2 was also significantly increased.Conclusion 1.A stable and reliable mouse model of H.suis infection was successfully established. 2. H. suis could induce the formation of lymphatic follicles in gastric mucosa.Obesity induced by high fat diet may promote the expression of IFN 纬 CXCL13 chain in leptin, and then promote the formation of gastric mucosal associated lymphoid tissue after H.suis infection.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2;R589.2

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