FAK及磷酸化FAK在尸检标本中冠状动脉粥样硬化局部的表达及意义

发布时间：2018-08-20 17:44

【摘要】：背景:动脉粥样硬化(Atherosclerosis,AS)发病率逐年升高,而由此引发的冠心病及近年来日益受到关注的急性冠脉综合征(Acute Coronary Syndromes,ACS)亦对健康造成威胁,严重者直接危害生命。随着相关研究的深入及扩展,整合素α5β1(Integrinα5β1)及其下游关键通路黏着斑激酶(Focal Adhesion Kinase,FAK)逐渐被纳入视线。最初研究表明FAK与与人类许多恶性肿瘤的发生、侵袭、转移及预后有着密切联系,如结肠、乳腺癌、胃癌、非小细胞肺癌等。FAK的生物学效应主要依赖于关键位点(Tyr397)的磷酸化而实现。另有文献报道,FAK在胚胎期血管生成及血管损伤后血管新生等修复过程中发挥重要作用,并且参与内皮细胞、平滑肌细胞的迁移及增殖等生物学过程。故推测其参与并促进动脉粥样硬化发生、发展。目的:本实验通过免疫组织化学的方法,检测FAK、Phospho FAK、CD131、CD68、actin在实验组(动脉粥样硬化组)、对照组(正常动脉组)中表达的情况。综合分析粥样硬化斑块FAK、Phospho FAK的表达情况与二者的相关性,及二者与AS发生、发展的关系。方法:标本选自近5年大连医科大学病理学与法医学教研室尸检标本100例。分别取左冠脉前降支、左旋支及右冠脉。主要实验手段采用免疫组织化学方法、弹力纤维染色法,并应用图像分析软件对图像进行分析及处理。以CD68与actin的表达定位FAK、Phospho FAK的表达情况。CD131标记新生血管,并计数新生血管的密度来检测FAK与血管新生的相关性。弹力纤维染色及NIH Scion Image(60.1)软件的应用判定冠状动脉狭窄程度。依据粥样硬化斑块的不同时期分为脂纹组及粥样硬化斑块组;依据EVG染色及软件分析结果,按狭窄程度将实验组冠脉分为4个亚组:分别为A组(≤25%)、B组(25%-50%)、C组(51%-75%)、D组(≥76%);依据病史及是否有急性心梗发作将病例分为ACS组、陈旧心梗组及正常组。结果:1)FAK与Phospho FAK均表达于内皮细胞胞浆,内膜间质细胞胞浆,巨噬细胞胞浆,平滑肌细胞胞浆,尤以迁移至内膜的平滑肌细胞表达高及钙化灶处。2)FAK在平滑肌细胞的表达强度与狭窄程度的关系:随着狭窄程度的升高而降低,两两比较分析P值均为0.000。Phospho FAK在平滑肌细胞的表达强度与狭窄程度关系呈现随狭窄程度升高而降低趋势,P值分别为:A组与B组0.011,A组与C组0.172,A组与D组0.000,B组与C组0.221,B组与D组0.000,C组与D组0.000。3)FAK与Phospho FAK的表达与斑块局部血管新生的关系局部血管新生的数量随二者的表达强度增加而增多,FAK与Phospho FAK与新生血管之间存在一定的正相关性,且具有统计学意义(RFAK=0.983,PFAK=0.000;RPhospho FAK=0.994,PPh FAK=0.000)。4)FAK与Phospho FAK在动脉粥样硬化斑块组平滑肌细胞的表达呈明显正相关性(R=0.904,P=0.000)。此外,FAK与Phospho FAK在非斑块底部平滑肌胞浆的表达高于斑块底部平滑肌,且FAK与Phospho FAK在内膜近内弹力板处间质的表达脂纹期高于粥样硬化斑块期。结论:1.FAK与Phospho FAK通过激活内皮细胞、促进平滑肌细胞迁移、诱导斑块局部血管新生等途径参与冠状动脉粥样硬化的发生、发展。2.FAK与Phospho FAK参与动脉粥样硬化局部钙化的发生。
[Abstract]:BACKGROUND: The incidence of atherosclerosis (AS) is increasing year by year. Coronary heart disease (CHD) and acute coronary syndrome (ACS), which have attracted more and more attention in recent years, also pose a threat to health and directly endanger life. With the deepening and expansion of relevant studies, integrin alpha 5 beta 1 (Integrin alpha 5 beta 1) Focal Adhesion Kinase (FAK), a key downstream pathway of FAK, has been gradually brought into view. Initial studies have shown that FAK is closely related to the occurrence, invasion, metastasis and prognosis of many human malignancies, such as colon, breast cancer, gastric cancer, non-small cell lung cancer, etc. The biological effects of FAK mainly depend on the key site (Tyr397). In addition, FAK has been reported to play an important role in the process of angiogenesis and angiogenesis after vascular injury in embryonic period. It is also involved in the migration and proliferation of endothelial cells and smooth muscle cells. Methods: The expression of FAK, Phospho FAK, CD131, CD68 and actin in experimental group (atherosclerosis group) and control group (normal artery group) were detected by histochemical method. The left anterior descending coronary artery, left circumflex coronary artery and right coronary artery were taken respectively. Immunohistochemistry, elastic fiber staining and image analysis software were used to analyze and process the images. Elastic fiber staining and NIH Scion Image (60.1) software were used to determine the degree of coronary artery stenosis. The coronary arteries of the experimental group were divided into four subgroups according to the degree of narrowness: group A (< 25%), group B (25% - 50%), group C (51% - 75%) and group D (> 76%). The patients were divided into ACS group, old myocardial infarction group and normal group according to the history and whether there was acute myocardial infarction attack. The expression of FAK in smooth muscle cells decreased with the increase of stenosis, and the P value was 0.000. The relationship between the expression of Phospho FAK in smooth muscle cells and the degree of stenosis was narrow. The expression of FAK and Phospho FAK and local angiogenesis of plaque increased with the increase of expression intensity of FAK and Phospho FAK, and the number of local angiogenesis of plaque increased with the increase of expression intensity of FAK and Phospho FAK and Phospho FAK. There was a positive correlation between FAK and Phospho FAK in atherosclerotic plaque (R = 0.904, P = 0.000). In addition, FAK and Phospho FAK were positively correlated in non-plaque basal smooth muscle cytoplasm. The expression of FAK and Phospho FAK in the stroma near the intimal elastic plate was higher than that in atherosclerotic plaque. Conclusion: 1. FAK and Phospho FAK participate in the pathogenesis and development of coronary atherosclerosis by activating endothelial cells, promoting the migration of smooth muscle cells and inducing local angiogenesis of plaque. .FAK and Phospho FAK participate in the occurrence of atherosclerotic local calcification.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.4

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