本文選題：慢性牙周炎 + 血管發(fā)生　； 參考：《第四軍醫(yī)大學》2014年碩士論文

【摘要】：研究背景： 炎癥的發(fā)生、發(fā)展是機體一系列復雜的病理變化過程，其中血管的變化在炎癥的進展過程中發(fā)揮有重要作用。血管系統(tǒng)的擴張可以運送炎癥細胞至病損處，并為組織提供氧和營養(yǎng)，同時為炎癥相關的多種分子提供底物。在組織中已存在的血管上，以出芽的方式形成新生血管的過程稱為血管發(fā)生。血管發(fā)生主要表現(xiàn)為組織中血管（主要是毛細血管）的擴張與增殖，在病理檢查可見血管直徑及數目的改變。血管發(fā)生與疾病的發(fā)展及預后有密切關系。 炎癥下血管發(fā)生的過程，是由于多種因素導致促進血管發(fā)生的因子的過表達產生的，在各種細胞因子和生長因子的調節(jié)下，組織內的分子信號可以發(fā)生發(fā)生復雜的相互作用。從而介導產生新血管形成過程中的一系列生理變化，如血管內皮細胞的增殖和遷移、血管基底膜和細胞外基質的降解及重塑、血管結構的重建等。其中血管內皮生長因子（vascular endothelial growth factor, VEGF）是作用于內皮細胞的強有力的生長因子。VEGF通過對血管內皮細胞的作用，，可提高血管的通透性，影響內皮細胞的增殖、分化和遷移，同時還可以通過介導破骨細胞和成骨細胞的遷移影響骨的改建過程；基質細胞衍生因子-1α（stromal cell-derived factor-1alpha, SDF-1α）在多種細胞的存活、生長和遷移中均發(fā)揮有重要作用，可與VEGF相互作用影響血管發(fā)生過程。 慢性牙周炎是由細菌侵入引發(fā)的，以牙周支持組織的慢性炎癥、牙周附著喪失和牙槽骨的破壞為特點的慢性炎癥性疾病，最終可導致牙齒的缺失。牙周組織中血管發(fā)生的狀況對于組織的健康和牙周炎癥的進展具有重要意義，但與慢性牙周炎發(fā)生發(fā)展有何關系，調控血管發(fā)生的分子及其作用機制如何，仍無明確報道。 研究目的： 研究血管密度的變化與慢性牙周炎牙周組織破壞程度之間的關系，以及血管發(fā)生因子VEGF、SDF-1α等在牙周組織中的表達變化，探討血管發(fā)生對牙周炎癥的影響及相關因素。 研究方法： 實驗一：慢性牙周炎牙齦組織中血管密度的變化 在就診的患者中，根據納入標準和排除標準，選取符合條件的慢性牙周炎組和健康對照組的受試患者，說明研究目的和意義，征得患者理解，并簽署知情同意書；臨床檢測記錄患者牙周的探診深度（probing depth, PD）和臨床附著喪失（clinicalattachment loss, CAL）程度；手術中獲取患牙牙齦組織，常規(guī)固定、包埋，制備組織切片；HE染色觀查組織形態(tài)，免疫組化方法檢測CD31的表達，光學顯微鏡下計數CD31陽性表達管腔數目，計算血管密度（microvessels density, MVD），統(tǒng)計學分析健康組與炎癥組血管密度的表達差異，及血管密度與PD和CAL之間的相關性。 實驗二：慢性牙周炎牙齦組織中血管發(fā)生因子的表達變化 標本同實驗一，通過免疫組化染色檢測VEGF和SDF-1α的表達，鏡下計數VEGF和SDF-1α表達的陽性細胞數目并統(tǒng)計分析，進而分析慢性牙周炎組織中血管發(fā)生因子表達量與PD和CAL的相關性；同法獲取患牙牙齦組織，提取組織RNA，PCR技術分析兩組間細胞因子CD31、VEGF和SDF-1α基因水平的表達差異。 研究結果： 實驗一： 1、組織HE染色可見，組織結構上牙齦由上皮層和固有層組成，健康對照組可見上皮釘突插入固有層中，固有層有少量炎癥細胞的浸潤而慢性牙周炎組牙齦可見明顯的上皮釘突明顯向固有層中增生、伸長，固有層中有炎癥細胞大量聚集，上皮輕度過角化。 2、免疫組化染色結果顯示，CD31表達于所有血管內皮細胞，CD31陽性的血管主要位于組織固有層中；組間統(tǒng)計分析結果顯示，在慢性牙周炎組MVD的表達明顯高于健康對照組（P＜0.05）。 3、Pearson回歸分析顯示，MVD與PD和CAL間存在直線正相關關系，且具有統(tǒng)計學意義（P＜0.05）。 實驗二： 1、免疫組化染色結果顯示，血管發(fā)生相關因子陽性表達主要位于牙齦組織的固有層。VEGF與SDF-1α主要表達于血管內皮細胞、間質細胞及部分炎癥細胞，且炎癥聚集處的表達明顯增多。兩種指標均顯示在固有層炎癥浸潤廣泛部位表達明顯增強。 2、免疫組化和Real Time-PCR定量分析均顯示，血管發(fā)生相關因子VEGF和SDF-1α在慢性牙周炎組的表達與健康對照組相比均明顯升高，有統(tǒng)計學差異（P＜0.05）。 3、VEGF和SDF-1α表達量與PD和CAL間存在直線正相關關系（P＜0.05）。 結論： 在慢性牙周炎中，牙周組織血管密度的變化與牙周炎癥狀態(tài)、牙周組織破壞程度關系密切，存在正相關關系；VEGF和SDF-1α等血管發(fā)生相關因子表達量與慢性牙周炎的炎癥程度成正相關，在炎癥的發(fā)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.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R781.42

【引證文獻】

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1 陳娟娟;哺乳期大鼠骨密度、正畸牙移動距離的變化及VEGF在哺乳期大鼠牙周組織中的表達[D];華北理工大學;2016年

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