主动吸烟对口腔黏膜上皮细胞NOD1信号通路的影响
发布时间：2019-02-15 05:45
【摘要】：第一部分构建经口主动吸烟动物模型[目的]吸烟对呼吸系统、消化系统、生殖系统、感官系统等多系统造成危害,可导致各种疾病的发生、发展。为了研究烟草与人类疾病的相关性,吸烟相关疾病模型的建立及完善越来越受到人们的关注,目前吸烟相关疾病模型主要包括临床样本吸烟模型、体外吸烟细胞模型和吸烟动物模型三类。但迄今为止,有关主动吸烟动物模型方面的研究鲜少,经口主动吸烟动物模型更未见国内外文献报道。本实验的目的是构建经口主动吸烟动物模型。[方法]Wistar大鼠20只,主动吸烟组和非吸烟组各10只,雌雄各半。主动吸烟组大鼠采用主动吸烟装置,建立大鼠经口主动吸烟模型。主动吸烟组大鼠每只连续经口吸烟3个月,每日两次,每次0.5小时。非吸烟组大鼠由经口装置吸空气,时间和频率与主动吸烟组相同。观察大鼠吸烟成瘾性、口腔黏膜外观的改变、神经行为学改变和体重的变化等。组化技术(HE染色)检查口腔黏膜的组织病理改变。[结果]主动吸烟组大鼠约2周时间吸烟成瘾,3个月时大体观可见口腔黏膜上皮颜色变为灰白色,质地粗糙,弹性降低。经统计学分析,体重较非吸烟组显著降低。口腔粘膜临床大体观及组织病理学检查显示,主动吸烟组大鼠口腔黏膜发生白斑,发生率为75%。[结论]本实验构建了经口主动吸烟动物(大鼠)模型,主动吸烟造成大鼠口腔黏膜大体和病理学改变,诱导口腔白斑的发生。第二部分 主动吸烟对大鼠口腔黏膜上皮细胞NOD1信号通路的影响[目的]口腔黏膜上皮细胞中NOD1信号通路是口腔黏膜固有免疫的重要组成部分,经本课题组前期体外实验和人体组织标本研究发现,吸烟可影响NOD1信号通路关键蛋白的表达,提示吸烟可抑制NOD1通路,降低口腔黏膜局部免疫。基于前期研究,本实验在构建经口主动吸烟动物模型的基础上,研究经口主动吸烟对口腔黏膜上皮细胞NOD1信号通路的影响。[方法]通过免疫印迹和免疫组化技术及图像分析法,对主动吸烟组与非吸烟组大鼠颊粘膜上皮细胞NOD1、RIP2、NF-κB、P-NF-κB (phospho-NF-κB,磷酸化NF-κB)蛋白表达进行检测,比较两组表达水平的差异。[结果]相对于非吸烟组,主动吸烟组NOD1和NF-κB的蛋白表达水平下降,RIP2、和P-NF-κB的蛋白表达水平上升。[结论]主动吸烟可导致口腔黏膜上皮细胞中NOD1信号通路中关键蛋白的改变,结果显示,主动吸烟对口腔黏膜上皮细胞中NOD1信号通路产生了影响。
[Abstract]:The first part is to construct the animal model of active smoking through mouth [objective] smoking causes harm to respiratory system, digestive system, reproductive system, sensory system and so on, which can lead to the occurrence and development of various diseases. In order to study the relationship between tobacco and human diseases, more and more attention has been paid to the establishment and improvement of smoking-related disease models. At present, the models of smoking-related diseases mainly include the smoking models of clinical samples. In vitro smoking cell model and smoking animal model. But so far, there are few studies on active smoking animal models, and there is no domestic and foreign literature on active smoking animal models. The purpose of this study was to establish an animal model of active smoking via mouth. [methods] Twenty Wistar rats were divided into active smoking group (n = 10) and non smoking group (n = 10). The active smoking model was established by using active smoking device in the active smoking group. Rats in active smoking group smoked twice a day for 0.5 hours twice a day for 3 months. The time and frequency of inhaling air in non-smoking group was the same as that in active smoking group. The changes of smoking addiction, oral mucosa appearance, neurobehavioral changes and body weight were observed. The histopathological changes of oral mucosa were examined by HE staining. [results] in the active smoking group, the rats were addicted to smoking for about 2 weeks. At 3 months, the color of oral mucosal epithelium became gray and white, the texture was rough, and the elasticity was decreased. By statistical analysis, the body weight was significantly lower than that of non-smoking group. The clinical gross and histopathological examination of oral mucosa showed that leukoplakia was found in oral mucosa of active smoking group (75%). [conclusion] the animal model of active oral smoking was established in this experiment. The oral mucosa gross and pathological changes were induced by active smoking, and the occurrence of oral leukoplakia was induced. Part II effects of active smoking on NOD1 signaling pathway in rat oral mucosal epithelial cells [objective] NOD1 signaling pathway is an important part of oral mucosal innate immunity in rats. Previous experiments in vitro and human tissue samples showed that smoking could affect the expression of key proteins in NOD1 signaling pathway, suggesting that smoking could inhibit NOD1 pathway and reduce local immunity of oral mucosa. Based on the previous studies, the effects of oral active smoking on NOD1 signaling pathway in oral epithelial cells were studied based on the establishment of oral active smoking animal model. [methods] NOD1,RIP2,NF- 魏 B P-NF- 魏 B (phospho-NF- 魏 B) in buccal mucosal epithelial cells of active smoking and non-smoking rats were studied by Western blot, immunohistochemistry and image analysis. The expression of phosphorylated NF- 魏 B was detected and the differences between the two groups were compared. [results] compared with non-smoking group, the expression of NOD1 and NF- 魏 B protein decreased in active smoking group, and the protein expression level of RIP2, and P-NF- 魏 B increased in active smoking group. [conclusion] active smoking can result in the change of key proteins in NOD1 signaling pathway in oral mucosal epithelial cells. The results show that active smoking has an effect on NOD1 signaling pathway in oral mucosal epithelial cells.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R781
本文编号：2423027
[Abstract]:The first part is to construct the animal model of active smoking through mouth [objective] smoking causes harm to respiratory system, digestive system, reproductive system, sensory system and so on, which can lead to the occurrence and development of various diseases. In order to study the relationship between tobacco and human diseases, more and more attention has been paid to the establishment and improvement of smoking-related disease models. At present, the models of smoking-related diseases mainly include the smoking models of clinical samples. In vitro smoking cell model and smoking animal model. But so far, there are few studies on active smoking animal models, and there is no domestic and foreign literature on active smoking animal models. The purpose of this study was to establish an animal model of active smoking via mouth. [methods] Twenty Wistar rats were divided into active smoking group (n = 10) and non smoking group (n = 10). The active smoking model was established by using active smoking device in the active smoking group. Rats in active smoking group smoked twice a day for 0.5 hours twice a day for 3 months. The time and frequency of inhaling air in non-smoking group was the same as that in active smoking group. The changes of smoking addiction, oral mucosa appearance, neurobehavioral changes and body weight were observed. The histopathological changes of oral mucosa were examined by HE staining. [results] in the active smoking group, the rats were addicted to smoking for about 2 weeks. At 3 months, the color of oral mucosal epithelium became gray and white, the texture was rough, and the elasticity was decreased. By statistical analysis, the body weight was significantly lower than that of non-smoking group. The clinical gross and histopathological examination of oral mucosa showed that leukoplakia was found in oral mucosa of active smoking group (75%). [conclusion] the animal model of active oral smoking was established in this experiment. The oral mucosa gross and pathological changes were induced by active smoking, and the occurrence of oral leukoplakia was induced. Part II effects of active smoking on NOD1 signaling pathway in rat oral mucosal epithelial cells [objective] NOD1 signaling pathway is an important part of oral mucosal innate immunity in rats. Previous experiments in vitro and human tissue samples showed that smoking could affect the expression of key proteins in NOD1 signaling pathway, suggesting that smoking could inhibit NOD1 pathway and reduce local immunity of oral mucosa. Based on the previous studies, the effects of oral active smoking on NOD1 signaling pathway in oral epithelial cells were studied based on the establishment of oral active smoking animal model. [methods] NOD1,RIP2,NF- 魏 B P-NF- 魏 B (phospho-NF- 魏 B) in buccal mucosal epithelial cells of active smoking and non-smoking rats were studied by Western blot, immunohistochemistry and image analysis. The expression of phosphorylated NF- 魏 B was detected and the differences between the two groups were compared. [results] compared with non-smoking group, the expression of NOD1 and NF- 魏 B protein decreased in active smoking group, and the protein expression level of RIP2, and P-NF- 魏 B increased in active smoking group. [conclusion] active smoking can result in the change of key proteins in NOD1 signaling pathway in oral mucosal epithelial cells. The results show that active smoking has an effect on NOD1 signaling pathway in oral mucosal epithelial cells.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R781
【参考文献】
相关期刊论文 前1条
1 李晓捷,高晶,孙忠人;宫内感染致早产鼠脑瘫动物模型制备及其鉴定的实验研究[J];中国康复医学杂志;2004年12期
，本文编号：2423027
本文链接：https://www.wllwen.com/yixuelunwen/kouq/2423027.html
最近更新
教材专著
