肥胖状态下牙周感染对小鼠牙周组织和脾脏组织中B-1a细胞的影响

发布时间：2018-04-24 06:31

本文选题：肥胖 + 牙周炎　；参考：《南方医科大学》2017年硕士论文

【摘要】：动物研究发现,B-1a细胞能在感染性炎症或慢性炎症疾病的固有免疫机制中起到保护作用,它能在病毒或细菌入侵的早期快速清除感染源。牙周炎与肥胖,可分别引起局部感染性炎症与全身慢性炎症状态,二者相关联的流行病学报道很多,但是它们之间的免疫机制还尚不明了。本研究建立肥胖复合牙周炎小鼠模型,基于B-1a细胞存在牙周病损局部和肥胖与牙周炎相关联的理论基础,对B-1a细胞在肥胖和牙周炎两种炎症状态中发挥的病理机制进行探索。第一章肥胖状态下牙周感染对小鼠牙周组织中B-1a细胞的影响1.目的本章主要探讨肥胖状态下牙周感染对小鼠牙周组织中B-1a细胞的表达水平及功能的影响,分析肥胖伴牙周感染小鼠牙周组织中B-1a细胞膜表面特异性蛋白CD5,及B-1a细胞分泌的白细胞介素-10(IL-10)炎症因子和抗Ⅰ胶原抗体(抗COLⅠ抗体)的蛋白和mRNA表达变化。2.方法动物模型分组:将24只小鼠随机分成两组,每组各12只。其中一组喂以高脂饲料,为肥胖组(high fat diet,HF),另一组喂以低脂饲料,为正常体重组(low fat diet,LF)。饲料诱导30周后,分别从肥胖组和正常体重组中随机抽取6只做牙周真性结扎处理(periodontitis,P),每组剩余的6只做假性结扎处理(control,C),结扎处理5天,随即产生牙周炎组(LFP组)、正常体重对照组(LFC组)、肥胖组(HFC组)、肥胖复合牙周炎组(HFP组)。免疫组化染色定量分析和实时定量PCR基因水平检测:通过实时定量PCR和免疫组化染色方法对牙周组织中CD5、抗COL Ⅰ抗体、IL-10的蛋白和mRNA表达水平进行定量检测。牙周组织中CD5、抗COL Ⅰ抗体、IL-10的表达水平分别与体重和牙周组织浸润的炎症细胞数之间的相关性分析:小鼠称重;牙周组织HE染色后对浸润的炎症细胞定量计数;将体重、炎症细胞数与三种蛋白之间进行Pearson相关性分析。3.结果牙周组织中B-1a细胞的mRNA及蛋白的表达水平:B-1a细胞相关三种蛋白的牙周炎主效应均有显著性,牙周炎组中的IL-10、CD5、抗COLⅠ抗体的mRNA和蛋白表达水平均显著高于对照组(P0.001)。肥胖和牙周炎对牙周组织中B-1a细胞功能的影响:CD5、IL-10、抗COLⅠ抗体的蛋白表达水平与炎症细胞数之间均呈显著正相关性(r0,P0.001);但三者与体重之间的相关性均不显著(P0.05)。4.结论B-1a细胞在牙周炎症早期表达上调,所分泌的抗COL Ⅰ抗体可以抵御细菌初期对牙周组织的破坏。但肥胖并未对炎性牙周组织中B-1a细胞的表达水平及功能产生显著性影响。第二章肥胖状态下牙周感染对小鼠脾脏组织中B-1a细胞的影响1.目的本章主要探讨肥胖状态下牙周感染对小鼠脾脏组织中B-1a细胞表达水平及功能的影响,分析肥胖伴牙周感染小鼠脾脏组织中B-1a细胞膜表面特异性蛋白CD5,及B-1a细胞分泌的抗COL Ⅰ抗体和IL-10炎症因子的蛋白表达变化。2.方法蛋白质印迹定量分析:用蛋白质印迹方法对脾脏组织中CD5、IL-10、抗COL Ⅰ抗体的蛋白表达水平进行定量检测。将脾脏组织中CD5、IL-10、抗COL Ⅰ抗体的表达水平分别与体重和牙周组织浸润的炎症细胞数之间进行Pearson相关性分析。3.结果脾脏组织中B-1a细胞的表达水平:B-1a细胞相关三种蛋白的肥胖主效应均有显著性,肥胖组中的CD5、IL-10、抗COLⅠ抗体的蛋白表达水平均显著高于正常体重组(P0.001)。肥胖和牙周炎对脾脏组织中B-1a细胞功能的影响:CD5、IL-10、抗COLⅠ抗体的蛋白表达水平与体重之间均呈显著正相关性(r0,P0.001),但三者与牙龈炎症细胞数之间的相关性均不显著(P0.05)。4.结论B-1a细胞在肥胖机体内表达上调,但其发挥的功能尚不明确。牙周感染并未对肥胖机体脾脏组织中B-1a细胞的表达产生显著性影响。
[Abstract]:Animal studies have found that B-1a cells can play a protective role in the inherent immune mechanism of infectious inflammation or chronic inflammatory diseases. It can quickly clear the source of infection in the early stages of virus or bacterial invasion. Periodontitis and obesity can cause local infection and systemic chronic inflammatory disease. The two related epidemiological reports are very important. Many, but the immune mechanism between them is still unknown. This study established a model of obese and complex periodontitis in mice. Based on the theoretical basis of the association between periodontal lesions and obesity and periodontitis, B-1a cells were explored in the two inflammatory states of B-1a cells in obesity and periodontitis. The first chapter was the obesity state. The effect of periodontal infection on B-1a cells in the periodontal tissue of mice 1. the purpose of this chapter is to investigate the effect of periodontal infection on the expression level and function of B-1a cells in the periodontal tissue of the mice, and to analyze the specific protein CD5 of the B-1a cell membrane in the periodontal tissues of the obese and periodontal infected mice, and the interleukocyte secreted by B-1a cells. -10 (IL-10) inflammatory factor and anti collagen antibody (anti COL I antibody) protein and mRNA expression change.2. method animal model group: 24 mice were randomly divided into two groups, 12 rats in each group. One group was fed with high fat diet, the obese group (high fat diet, HF), the other group fed with low fat diet, for the normal weight group (low fat diet, low). After 30 weeks of induction, 6 patients were randomly selected from the obese group and the normal weight group for periodontitis (P). The remaining 6 of each group were treated with pseudoligature (control, C) and ligation for 5 days. Then the periodontitis group (group LFP), the normal weight control group (group LFC), the obese group (group HFC), and the obesity complex periodontitis group (group HFP). Quantitative analysis of immunohistochemical staining and real-time quantitative PCR gene level detection: quantitative detection of CD5, anti COL I antibody, IL-10 protein and mRNA expression level in periodontal tissues by real-time quantitative PCR and immunohistochemical staining. The expression of CD5, anti COL I antibody, and IL-10 expression level in periodontal tissues with body weight and periodontal tissue infiltration respectively Correlation analysis between the number of inflammatory cells: weighing in mice; quantitative counting of infiltrating inflammatory cells after HE staining in periodontal tissue; Pearson correlation analysis between body weight, number of inflammatory cells and three kinds of proteins; the mRNA and protein of B-1a cells in periodontal tissues were reached: the periodontitis owner of three kinds of protein related proteins in the periodontal tissue. The effect of IL-10, CD5, anti COL I antibody in the periodontitis group was significantly higher than that of the control group (P0.001). The effect of obesity and periodontitis on the function of B-1a cells in periodontal tissue: the protein expression level of CD5, IL-10, anti COL I antibody and the number of inflammatory cells was significantly positive correlation (R0, P0.001). The correlation between the three and the weight was not significant (P0.05).4. conclusion the expression of B-1a cells was up-regulated in the early periodontitis, and the secreted anti COL I antibody could resist the early destruction of periodontal tissue. But obesity did not have a significant effect on the expression level and function of B-1a cells in the inflammatory periodontitis. The second chapter of obesity was in the state of obesity. The effect of periodontal infection on the B-1a cells in the spleen tissue of mice 1. the purpose of this chapter is to investigate the effect of periodontal infection on the expression and function of B-1a cells in the spleen tissues of the mice, and to analyze the specific protein CD5 of the B-1a cell membrane in the spleen tissues of the obese and periodontal infected mice, and the anti COL I anti B-1a cells secreted by B-1a cells. Protein expression changes in body and IL-10 inflammatory factors.2. method of Western blot quantitative analysis: the protein expression level of CD5, IL-10, anti COL I antibody in spleen tissues was quantified by Western blot. The expression of CD5, IL-10, anti COL I antibody in the spleen tissues was divided with body weight and infiltration of periodontitis. Pearson correlation analysis between the numbers of.3. results in the expression level of B-1a cells in the spleen tissue: the main effect of obesity on three proteins related to B-1a cells was significant. The protein expression level of CD5, IL-10, anti COL I antibody in the obese group was significantly higher than that of the normal weight group (P0.001). The B-1a cell work in the spleen tissues of obesity and periodontitis Effect: the protein expression level of CD5, IL-10, anti COL I antibody showed significant positive correlation with body weight (R0, P0.001), but the correlation between the three and the number of gingivitis cells was not significant (P0.05).4. conclusion B-1a cells were up regulated in the body of obesity, but the function of the cells was not clear. Periodontal infection was not on the obese body. The expression of B-1a cells in spleen tissue has a significant effect.

【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R781.4

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