NLRP3信号通路在不同炎症状态下对牙周炎发生发展的作用研究

发布时间：2018-06-28 11:03

  本文选题：慢性炎症 + 配胖　； 参考：《南方医科大学》2017年博士论文

【摘要】：1.研究背景及目的慢性炎症是一系列慢性疾病,如肥胖、糖尿病等的共同病理机制。肥胖机体以长期的低度慢性系统性炎症及免疫反应异常、胰岛素抵抗为重要特征,使其易感于重大代谢疾病包括糖尿病等。肥胖是糖尿病的重要病因,二者呈“孪生”流行趋势,且均可促进牙周炎的发展。而关于NLRP3信号通路在其中所起的关键作用的研究匮乏。本研究目的为探讨肥胖及Ⅱ型糖尿病导致的系统性炎症状态对牙周组织炎症的影响及NLRP3信号通路在其中所起的关键作用。2.研究方法(1)饮食诱导16周牙周结扎10天建立肥胖小鼠牙周炎模型,记为正常饮食组、正常饮食结扎组、高脂饮食组、高脂饮食结扎组。(2)体内通过RT-PCR和IHC的方法检测小鼠牙龈组织内NLRP3信号通路(NLRP3,capase1,IL-1β,NFκb)及巨噬细胞表面标记物F4/80、巨噬细胞趋化因子MCP1的表达情况。(3)体外分离培养不同饮食背景的小鼠BMDMs并对其进行LPS刺激,通过RT-PCR及ICC检测巨噬细胞内NLRP3信号通路的表达。(4)收集人类健康组、慢性牙周炎组及慢性牙周炎伴Ⅱ型糖尿病组患者牙龈组织,体内通过RT-PCR及IHC检测牙龈组织内NLRP3信号通路的表达。体外分离培养人牙龈上皮细胞并对其进行高糖及LPS刺激实验,通过RT-PCR及ICC检测上皮细胞内NLRP3信号通路的表达。3.研究结果(1)高脂饲料诱导16w小鼠的体重及血糖明显高于正常饲料组。小鼠牙槽骨吸收:牙周炎组显著高于非牙周炎组,高脂饮食组显著高于正常饮食组。血清胰岛素及炎症因子TNF-α,IL-1β,IL-6,IL-10表达水平:高脂饮食组显著高于正常饮食组。(2)小鼠牙龈组织中NLRP3,Caspasel,IL-1β及NFκb的mRNA及蛋白质表达水平:牙周炎组显著高于非牙周炎组。(3)小鼠牙龈组织中F4/80及MCP1的mRNA及蛋白质表达水平:牙周炎组显著高于非牙周炎组,高脂饮食组显著低于正常饮食组。BMDMs中NLRP3信号通路因子的mRNA及蛋白质表达水平:LPS刺激组显著高于非刺激组,高脂饮食组显著低于正常饮食组。(4)人牙龈组织中NLRP3信号通路因子的mRNA及蛋白质表达水平:在伴有炎症及Ⅱ型糖尿病的牙龈组织中表达显著性升高。HGECs中NLRP3信号通路因子的mRNA及蛋白质表达水平在高糖刺激后显著升高。4.结论我们发现,肥胖、Ⅱ型糖尿病的机体处于慢性炎症状态均加重了牙周炎的发生发展。肥胖导致感染的牙周组织内巨噬细胞数量减少及活性降低,引起牙周固有免疫的异常,最终加重了牙周炎的局部炎症状况。而Ⅱ型糖尿病患者的高糖状态可能通过牙龈上皮细胞NLRP3信号通路的高表达而加重慢性牙周炎的发生发展。
[Abstract]:1. Background and objective chronic inflammation is a common pathological mechanism of a series of chronic diseases such as obesity and diabetes. Obesity is characterized by chronic low degree chronic systemic inflammation, abnormal immune response and insulin resistance, which makes it susceptible to major metabolic diseases, including diabetes, etc. Obesity is an important cause of diabetes, both of which are "twin" epidemic trend, and can promote the development of periodontitis. However, there is little research on the key role of NLRP3 signaling pathway. The aim of this study was to investigate the effects of systemic inflammation induced by obesity and type 2 diabetes on periodontal inflammation and the key role of NLRP3 signaling pathway. Methods (1) Obesity mouse periodontitis model was established by 16 weeks periodontal ligation induced by diet for 10 days, which was recorded as normal diet group, normal diet ligation group, high fat diet group, and high fat diet group. High fat diet ligation group. (2) the expression of NLRP3 signal pathway (NLRP3 signal pathway), macrophage surface marker F4 / 80 and macrophage chemokine MCP1 in vitro were detected by RT-PCR and IHC in mice gingival tissue. (3) the expression of NLRP3 signal pathway (NLRP3) and macrophage surface marker F4 / 80, MCP1 were detected in vitro. Jing mice BMDMs were stimulated by LPS. The expression of NLRP3 signal pathway in macrophages was detected by RT-PCR and ICC. (4) gingival tissues were collected from healthy people, chronic periodontitis patients and chronic periodontitis patients with type 鈪，

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