慢性牙周炎中血管发生的表达变化及相关性分析

发布时间：2018-06-19 22:51

本文选题：慢性牙周炎 + 血管发生　；参考：《第四军医大学》2014年硕士论文

【摘要】：研究背景：炎症的发生、发展是机体一系列复杂的病理变化过程，其中血管的变化在炎症的进展过程中发挥有重要作用。血管系统的扩张可以运送炎症细胞至病损处，并为组织提供氧和营养，同时为炎症相关的多种分子提供底物。在组织中已存在的血管上，以出芽的方式形成新生血管的过程称为血管发生。血管发生主要表现为组织中血管（主要是毛细血管）的扩张与增殖，在病理检查可见血管直径及数目的改变。血管发生与疾病的发展及预后有密切关系。炎症下血管发生的过程，是由于多种因素导致促进血管发生的因子的过表达产生的，在各种细胞因子和生长因子的调节下，组织内的分子信号可以发生发生复杂的相互作用。从而介导产生新血管形成过程中的一系列生理变化，如血管内皮细胞的增殖和迁移、血管基底膜和细胞外基质的降解及重塑、血管结构的重建等。其中血管内皮生长因子（vascular endothelial growth factor, VEGF）是作用于内皮细胞的强有力的生长因子。VEGF通过对血管内皮细胞的作用，，可提高血管的通透性，影响内皮细胞的增殖、分化和迁移，同时还可以通过介导破骨细胞和成骨细胞的迁移影响骨的改建过程；基质细胞衍生因子-1α（stromal cell-derived factor-1alpha, SDF-1α）在多种细胞的存活、生长和迁移中均发挥有重要作用，可与VEGF相互作用影响血管发生过程。慢性牙周炎是由细菌侵入引发的，以牙周支持组织的慢性炎症、牙周附着丧失和牙槽骨的破坏为特点的慢性炎症性疾病，最终可导致牙齿的缺失。牙周组织中血管发生的状况对于组织的健康和牙周炎症的进展具有重要意义，但与慢性牙周炎发生发展有何关系，调控血管发生的分子及其作用机制如何，仍无明确报道。研究目的：研究血管密度的变化与慢性牙周炎牙周组织破坏程度之间的关系，以及血管发生因子VEGF、SDF-1α等在牙周组织中的表达变化，探讨血管发生对牙周炎症的影响及相关因素。研究方法：实验一：慢性牙周炎牙龈组织中血管密度的变化在就诊的患者中，根据纳入标准和排除标准，选取符合条件的慢性牙周炎组和健康对照组的受试患者，说明研究目的和意义，征得患者理解，并签署知情同意书；临床检测记录患者牙周的探诊深度（probing depth, PD）和临床附着丧失（clinicalattachment loss, CAL）程度；手术中获取患牙牙龈组织，常规固定、包埋，制备组织切片；HE染色观查组织形态，免疫组化方法检测CD31的表达，光学显微镜下计数CD31阳性表达管腔数目，计算血管密度（microvessels density, MVD），统计学分析健康组与炎症组血管密度的表达差异，及血管密度与PD和CAL之间的相关性。实验二：慢性牙周炎牙龈组织中血管发生因子的表达变化标本同实验一，通过免疫组化染色检测VEGF和SDF-1α的表达，镜下计数VEGF和SDF-1α表达的阳性细胞数目并统计分析，进而分析慢性牙周炎组织中血管发生因子表达量与PD和CAL的相关性；同法获取患牙牙龈组织，提取组织RNA，PCR技术分析两组间细胞因子CD31、VEGF和SDF-1α基因水平的表达差异。研究结果：实验一： 1、组织HE染色可见，组织结构上牙龈由上皮层和固有层组成，健康对照组可见上皮钉突插入固有层中，固有层有少量炎症细胞的浸润而慢性牙周炎组牙龈可见明显的上皮钉突明显向固有层中增生、伸长，固有层中有炎症细胞大量聚集，上皮轻度过角化。 2、免疫组化染色结果显示，CD31表达于所有血管内皮细胞，CD31阳性的血管主要位于组织固有层中；组间统计分析结果显示，在慢性牙周炎组MVD的表达明显高于健康对照组（P＜0.05）。 3、Pearson回归分析显示，MVD与PD和CAL间存在直线正相关关系，且具有统计学意义（P＜0.05）。实验二： 1、免疫组化染色结果显示，血管发生相关因子阳性表达主要位于牙龈组织的固有层。VEGF与SDF-1α主要表达于血管内皮细胞、间质细胞及部分炎症细胞，且炎症聚集处的表达明显增多。两种指标均显示在固有层炎症浸润广泛部位表达明显增强。 2、免疫组化和Real Time-PCR定量分析均显示，血管发生相关因子VEGF和SDF-1α在慢性牙周炎组的表达与健康对照组相比均明显升高，有统计学差异（P＜0.05）。 3、VEGF和SDF-1α表达量与PD和CAL间存在直线正相关关系（P＜0.05）。结论：在慢性牙周炎中，牙周组织血管密度的变化与牙周炎症状态、牙周组织破坏程度关系密切，存在正相关关系；VEGF和SDF-1α等血管发生相关因子表达量与慢性牙周炎的炎症程度成正相关，在炎症的发展中具有重要作用。
[Abstract]:Research background:
Inflammation, development is a complex series of pathological changes in the body, in which vascular changes play an important role in the progression of inflammation. The expansion of the vascular system can transport inflammatory cells to the lesion, provide oxygen and nutrition for tissue, and provide substrates for various molecules in inflammation. On the blood vessels, the process of forming new blood vessels in the way of buds is called angiogenesis. Angiogenesis is mainly manifested by the expansion and proliferation of blood vessels in the tissue (mainly capillary), and the changes in the diameter and number of blood vessels are seen in the pathological examination.
The process of vasculogenesis under inflammation is caused by a variety of factors that lead to the overexpression of factors that promote angiogenesis. Under the regulation of various cytokines and growth factors, molecular signals in the tissue can occur complex interactions, which mediate a series of physiological changes in the formation of new vessels, such as intravascular blood vessels. The proliferation and migration of skin cells, degradation and remodeling of vascular basement membrane and extracellular matrix, reconstruction of vascular structure, and vascular endothelial growth factor (vascular endothelial growth factor, VEGF) is a strong growth factor.VEGF acting on endothelial cells through the action of the vascular endothelial cells, which can improve the permeability of blood vessels. The proliferation, differentiation and migration of endothelial cells can also affect the process of bone remodeling by mediating the migration of osteoclasts and osteoblasts; the matrix cell derived factor -1 alpha (stromal cell-derived factor-1alpha, SDF-1 a) plays an important role in the survival, growth and migration of a variety of cells and can interact with VEGF. It affects the process of vasculogenesis.
Chronic periodontitis is a chronic inflammatory disease characterized by chronic inflammation of the periodontal supporting tissue, loss of periodontal attachment and destruction of alveolar bone in periodontal tissue, which can eventually lead to the loss of teeth. The status of angiogenesis in periodontal tissue is of great significance to the health of the tissues and the progress of periodontitis, but with chronic teeth. What is the relationship between the occurrence and development of periodontitis, and how the molecules regulating angiogenesis and their mechanisms of action are still not reported clearly.
The purpose of the study is:
To study the relationship between the changes of blood vessel density and the degree of periodontal tissue damage in chronic periodontitis, as well as the changes in the expression of angiogenic factor VEGF, SDF-1 alpha in periodontium and the influence of angiogenesis on periodontitis and related factors.
Research methods:
Experiment 1: changes of vascular density in gingival tissue of chronic periodontitis
According to the inclusion criteria and exclusion criteria, the subjects of the eligible chronic periodontitis group and the healthy control group were selected to illustrate the purpose and significance of the study, to obtain the understanding of the patients, and to sign the informed consent; the clinical detection records of the patients' probing depth (PD) and the loss of clinical attachment (clinicalat The degree of tachment loss, CAL); during the operation, the gingival tissues of the affected teeth were obtained, the routine fixation, the embedding, and the preparation of tissue sections; the HE staining was used to examine the tissue morphology, the immunohistochemical method was used to detect the expression of CD31, the number of CD31 positive expressions of the lumen was counted under the optical microscope, the density of the blood vessel (microvessels density, MVD) was calculated, and the health group and inflammation were analyzed statistically. The difference of the expression of vessel density and the correlation between vascular density and PD and CAL in the disease group.
Experiment two: expression of vascular endothelial growth factor in gingival tissue of chronic periodontitis
The expression of VEGF and SDF-1 alpha was detected by immunohistochemical staining. The number of positive cells expressed by VEGF and SDF-1 alpha was counted under the microscope. The correlation between the expression of angiogenic factor and PD and CAL in the tissues of chronic periodontitis was analyzed, and the two groups of RNA and PCR techniques were obtained by the same method. The difference of cytokine CD31, VEGF and SDF-1 alpha gene expression levels.
The results of the study:
Experiment 1:
1, the tissue HE staining showed that the gingiva was composed of epithelial layer and lamina propria on the tissue structure. In the healthy control group, the epithelial nail process was inserted into the lamina propria, and a small amount of inflammatory cells were infiltrated in the lamina propria, and the gingiva of the chronic periodontitis group showed obvious epithelial nailed process to the lamina propria. The skin is light through keratinization.
2, immunohistochemical staining showed that CD31 was expressed in all vascular endothelial cells, and the CD31 positive blood vessels were mainly in the lamina propria. The statistical analysis showed that the expression of MVD in the chronic periodontitis group was significantly higher than that in the healthy control group (P < 0.05).
3, Pearson regression analysis showed that there was a positive linear correlation between MVD and PD and CAL, and was statistically significant (P < 0.05).
Experiment two:
1, immunohistochemical staining showed that the positive expression of angiogenesis related factors was mainly expressed in the lamina propria.VEGF and SDF-1 alpha mainly in vascular endothelial cells, interstitial cells and some inflammatory cells, and the expression of inflammatory aggregation was obviously increased. The two indicators showed obvious expression in the extensive region of propria infiltration. Enhance.
2, immunohistochemistry and Real Time-PCR quantitative analysis showed that the expression of angiogenesis related factors VEGF and SDF-1 alpha in the chronic periodontitis group was significantly higher than that in the healthy control group, and there was a statistical difference (P < 0.05).
3, there was a positive linear correlation between the expression of VEGF and SDF-1 and PD and CAL (P < 0.05).
Conclusion:
In chronic periodontitis, the changes of vascular density in periodontal tissue are closely related to periodontal inflammation and periodontal tissue damage, and there is a positive correlation between the expression of VEGF and SDF-1 alpha angiogenesis related factors and the degree of inflammation in chronic periodontitis, which plays an important role in the development of inflammation.
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.42

【引证文献】