HIF-1α和VEGF-c在人炎症牙髓组织中表达的研究

发布时间：2018-04-28 14:42

本文选题：牙髓炎 + 氧代谢　；参考：《暨南大学》2015年硕士论文

【摘要】：背景及目的:牙髓炎是由细菌作为始动因子引起牙髓组织的慢性感染性疾病。乏氧诱导因子-1(hypoxia inducible factor-1,HIF-1)作为细胞在乏氧情况下由激活基因编码的转录因子。血管内皮生长因子(vascular endothelial growth factor,VEGF)是一种高度特异性的细胞因子,具有促进血管内皮细胞分裂、诱发血管形成及增加血管通透性的功能。在乏氧状态下,HIF-1能够诱导而转录激活VEGF,其作为HIF-1α的主要靶基因之一。目前研究发现,HIF-1α和VEGF在肿瘤血管生成之间存在着密切关系。至今为止,HIF-1α和VEGF-c在牙髓炎中的作用研究甚少。本课题主要研究目的是观察炎性牙髓组织中HIF-1α和VEGF-c表达与分布情况,分析牙髓组织在不同状态下的缺氧情况及其程度。探讨HIF-1α和VEGF-c调控牙髓组织炎症进程中的作用,研究结果将为阐明牙髓炎的氧代谢机制以及牙髓炎的靶向治疗提供理论依据。方法:临床随机收集年龄在20~50岁之间患者的牙髓组织,其中男性18名,女性22名,平均年龄32.9±11.4岁。经统计学处理分析各组之间在年龄、性别上差异无统计学意义(P0.05),具有可比性。根据牙髓组织病理学分类进行如下四类分组:正常牙髓组、深龋牙髓组、慢性牙髓炎组以及急性牙髓炎组共40例。所有牙髓标本来自因正畸需要拔除或无法保留的第三磨牙。牙髓组织标本于2%戊二醛溶液中固定48 h以上,制成颊舌向5μm厚度连续切片。通过HE染色方法,观察各组牙髓标本的组织学改变并评分;应用免疫组织化学染色法,观察HIF-1α和VEGF-c在人牙髓组织中的表达及分布、并计算阳性细胞密度。应用SPSS 16.0统计学软件对所有数据进行分析处理:1)用Kruskal-Wallis H秩和检验比较不同组间牙髓组织炎症程度评分;2)用Kruskal-Wallis H秩和检验比较各组间牙髓HIF-1α和VEGF-c阳性细胞密度;3)用Pearson相关分析,评估各组间牙髓炎症程度评分与HIF-1α、VEGF-c阳性细胞密度之间的相关性;4)通过Pearson相关分析,评估不同组间HIF-1α和VEGF-c表达水平之间的相关性。结果:1.牙髓组织学观察结果1)正常牙髓组与深龋牙髓组中,无明显炎性细胞浸润和炎性渗出物。2)与正常牙髓组相比,慢性牙髓炎组的牙髓组织出现中等程度的炎症细胞浸润。3)与慢性牙髓炎组相比,急性牙髓炎组表现为更明显的炎性细胞浸润。4)与正常牙髓组与深龋牙髓组相比,炎症牙髓组织的炎症程度评分显著升高(P0.01)。其中,急性牙髓炎组的炎症程度评分明显高于慢性牙髓炎(P0.01)。2.牙髓组织中HIF-1α与VEGF-c免疫组化染色结果HIF-1α表达主要定位于成牙本质细胞、成纤维细胞、血管内皮细胞及部分炎症细胞。正常牙髓组与深龋牙髓组中,牙髓组织几乎没有HIF-1α阳性表达出现。与正常牙髓组与深龋牙髓组相比较,慢性牙髓炎组中HIF-1α阳性细胞密度显著升高(P0.01);且急性牙髓炎组HIF-1α阳性细胞密度显著高于慢性牙髓炎组(P0.01)。VEGF-c表达主要定位于成牙本质细胞、成纤维细胞、血管内皮细胞,巨噬细胞和浆细胞。正常牙髓组与深龋牙髓组中,牙髓组织见少量VEGF-c阳性表达出现。与正常牙髓组与深龋牙髓组相比较,急性牙髓炎组中VEGF-c阳性细胞密度显著升高(P0.01);且慢性牙髓炎组VEGF-c阳性细胞密度显著高于急性牙髓炎组(P0.01)。3.HIF-1α和VEGF-c阳性细胞密度与牙龈组织炎症程度评分的相关性分析1)通过Pearson相关分析,不同组牙髓组织中HIF-1α阳性细胞密度与炎症程度评分存在正的直线相关关系(P0.01)。2)通过Pearson相关分析,不同组牙髓组织中VEGF-c阳性细胞密度与炎症程度评分存在正的直线相关关系(P0.01)。4.HIF-1α和VEGF-c表达水平的相关性分析经Pearson相关分析,炎症牙髓组织中HIF-1α阳性细胞密度与VEGF-c阳性细胞密度密切相关,且存在正的直线相关关系(P0.01)。结论:1.与正常牙髓组、深龋牙髓组比较,炎症的牙髓组织HIF-1α和VEGF-c的表达明显升高。2.HIF-1α和VEGF-c的表达量与牙髓炎症程度存在明显的相关性。结果提示HIF-1α和VEGF-c可能参与牙髓炎的炎症反应,其具体的作用机制仍需要进一步的研究。
[Abstract]:Background and purpose: pulpitis is a chronic infectious disease of dental pulp tissue caused by bacteria as a starting factor. Hypoxia inducible factor -1 (hypoxia inducible factor-1, HIF-1) is a transcription factor encoded by activated genes in hypoxic conditions. Vascular endothelial growth factor (vascular endothelial growth factor, VEGF) is a kind of height. Specific cytokine has the function of promoting vascular endothelial cell division, inducing angiogenesis and increasing vascular permeability. In hypoxic state, HIF-1 can induce and transcriptional activation of VEGF, which is one of the main target genes of HIF-1 alpha. Current research has found that there is a close relationship between HIF-1 alpha and VEGF in tumor angiogenesis. So far, there is little research on the role of HIF-1 alpha and VEGF-c in pulpitis. The main purpose of this study is to observe the expression and distribution of HIF-1 alpha and VEGF-c in inflammatory pulp tissues, to analyze the condition of the anoxia and the extent of the dental pulp tissue in different states. The role of HIF-1 alpha and VEGF-c in the regulation of the inflammatory process of dental pulp tissue is discussed, and the results will be studied. To provide a theoretical basis for elucidating the oxygen metabolism mechanism of pulpitis and the targeting therapy of pulpitis. Methods: the dental pulp tissues of patients aged 20~50 were randomly collected, including 18 men and 22 women, with an average age of 32.9 + 11.4 years. There was no statistically significant difference in age between groups (P0.05) by statistical analysis. According to the pathological classification of dental pulp tissue, the following four groups of groups were divided: normal pulp group, deep caries pulp group, chronic pulpitis group and acute pulpitis group with 40 cases. All dental pulp specimens were extracted or unreserved third molar teeth. The dental pulp tissue was fixed in 2% glutaraldehyde solution more than 48 h, made of dental pulp tissue. The buccal tongue to 5 m thickness continuous slice. Through the HE staining method, the histological changes of the dental pulp specimens were observed and the scores were observed. The expression and distribution of HIF-1 alpha and VEGF-c in the human dental pulp tissues were observed and the density of the positive cells was calculated by immunohistochemical staining. All data were analyzed with the SPSS 16 system software: 1) K Ruskal-Wallis H rank sum test compared the degree of inflammation in dental pulp tissue between different groups; 2) the density of HIF-1 alpha and VEGF-c positive cells in dental pulp of each group was compared with Kruskal-Wallis H rank and test; 3) the correlation between the degree of inflammation of dental pulp and the density of HIF-1 A and VEGF-c positive cells was evaluated by Pearson correlation analysis; 4) through Pearson. Correlation analysis, evaluation of the correlation between HIF-1 alpha and VEGF-c expression levels between different groups. Results: 1. the results of dental pulp histology were 1) in the normal pulp group and the deep caries pulp group, there was no obvious inflammatory cell infiltration and inflammatory exudate.2) and the pulp tissue of the chronic pulpitis group had moderate inflammatory cell infiltration compared with the normal pulp group. 3) compared with the chronic pulpitis group, the acute pulpitis group showed a more obvious inflammatory cell infiltration.4). Compared with the normal dental pulp group and the deep caries pulp group, the inflammatory degree score of the inflammatory pulp tissue was significantly higher (P0.01). Among them, the severity score of the acute pulpitis group was significantly higher than that of the chronic pulpitis (P0.01).2. pulp tissue of HIF-1 a. The expression of HIF-1 alpha in VEGF-c immunohistochemical staining was mainly located in odontoblast, fibroblasts, vascular endothelial cells and some inflammatory cells. In the normal pulp group and the deep caries pulp group, there was almost no positive expression of HIF-1 alpha in the pulp tissue. Compared with the normal pulp group and the deep caries pulp group, the HIF-1 alpha in the chronic pulpitis group was in the group of the normal pulp and the deep caries pulp group. The density of positive cells increased significantly (P0.01), and the density of HIF-1 alpha positive cells in the acute pulpitis group was significantly higher than that in the chronic pulpitis group (P0.01). The expression of.VEGF-c was mainly located in odontoblasts, fibroblasts, vascular endothelial cells, macrophages and plasma cells. In the normal dental pulp group and the deep caries pulp group, a small amount of VEGF-c positive was found in the pulp tissue. Compared with the normal dental pulp group and the deep carious pulp group, the density of VEGF-c positive cells in the acute pulpitis group increased significantly (P0.01), and the density of VEGF-c positive cells in the chronic pulpitis group was significantly higher than that of the acute pulpitis group (P0.01), the correlation between the density of.3.HIF-1 alpha and VEGF-c positive cells and the grade of gingival tissue inflammation was 1). After Pearson correlation analysis, there was a positive linear correlation between the density of HIF-1 alpha positive cells in the dental pulp tissues and the degree of inflammation (P0.01).2 in different groups of dental pulp tissues. Through Pearson correlation analysis, there was a positive linear correlation between the density of VEGF-c positive cells in the dental pulp tissues and the degree of inflammation (P0.01).4.HIF-1 A and VEGF-c expression levels in different groups of dental pulp tissues. The correlation analysis was analyzed by Pearson correlation analysis. The density of HIF-1 alpha positive cells in inflammatory pulp tissues was closely related to the density of VEGF-c positive cells, and there was a positive linear correlation (P0.01). Conclusion: 1. the expression of HIF-1 alpha and VEGF-c in the pulp tissues of the normal dental pulp group and the deep carious pulp group increased the table of.2.HIF-1 A and VEGF-c. The results suggest that HIF-1 alpha and VEGF-c may be involved in the inflammatory response of pulpitis, and the specific mechanism of its action still needs further study.

【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R781.31

【共引文献】