人冠状动脉粥样病灶内CD40L水平与病灶结构变化的关系

发布时间：2018-08-17 10:14

【摘要】：目的:为了探讨CD40-CD40L信号在人冠状动脉粥样硬化(atherosclerosis,AS)病灶发生发展的作用,本课题进行了人冠状动脉AS病灶结构变化的观察和病灶内CD40L、MMP-9表达水平的检测。方法:收集人冠状动脉AS血管60例作为实验组,按照病理分期分为四组:脂纹期5例,纤维斑块期19例,粥样斑块期18例,继发改变期18例;以无病变冠状动脉12例为对照组。常规HE染色观察两组人群冠状动脉的组织结构并用图像分析软件测量病灶结构的相关指标,免疫组化(Immunohistochemistry,IHC)SP两步法检测CD40L、MMP-9蛋白表达水平,逆转录-实时荧光定量PCR(Reverse transcription-quantitative real time PCR,RT-qPCR)法检测CD40L、MMP-9 mRNA表达水平,分析CD40L、MMP-9表达水平与冠脉AS病灶结构变化之间的关系。结果:1.对照组冠脉管壁薄,内膜、中膜、外膜各层结构厚度均匀一致,内膜平滑完整。实验组肉眼呈从脂纹期到继发病变期的不同变化,病变使冠脉出现不同程度的管壁增厚和管腔狭窄,部分病灶内可见钙化、出血、血栓形成等;光镜下轻者仅见泡沫细胞增生,血管壁厚度及管腔无明显改变;重者出现明显粥样坏死灶,甚至斑块内出血或血栓形成,血管壁显著增厚,管腔明显狭窄。2.对照组血管壁内未见CD40L表达,实验组CD40L表达水平明显增高,阳性表达主要分布于AS病灶肩区、基底部、纤维帽区域及坏死灶周围的泡沫细胞及炎细胞胞浆,CD40L平均光密度值测定显示实验组显著高于对照组(P0.05),且实验组内各期病变CD40L水平具有显著性差异(P0.05)。3.对照组血管壁可见MMP-9蛋白少量表达于中膜平滑肌细胞及内膜泡沫细胞的胞浆;实验组冠脉AS病灶内可见MMP-9蛋白大量表达于泡沫细胞及炎细胞,以粥样斑块肩区、基底部、纤维帽区域及坏死灶周围明显,另可见少量表达于中膜平滑肌细胞。平均光密度值测定显示,实验组MMP-9水平显著高于对照组(P0.05);且不同病变阶段的病灶内MMP-9表达水平存在显著性差异(P0.05)。4.RT-qPCR法检测冠脉组织中CD40L、MMP-9 mRNA水平,结果显示实验组CD40L水平高于对照组(P0.05),进一步比较仅继发病变期高于对照组(P0.05);MMP-9 mRNA实验组与对照组比较,无论组间或组内,差异均无统计学意义(P0.05)。5.蛋白CD40L与MMP-9的表达水平呈正相关,且二者与内膜最厚处厚度、坏死灶厚度、血管狭窄程度呈正相关(P0.05),与纤维帽厚度分数有一定的负相关趋势。结论:1.人冠脉AS病灶内CD40L表达水平明显增强。2.人冠脉AS病灶内MMP-9高表达,并与CD40L表达水平正相关,与病灶结构指标变化有关。3.人冠脉AS病灶内CD40L可能促进MMP-9表达,使病灶内细胞外基质(extracellular matrix,ECM)降解加速,最终导致病灶结构发生纤维帽变薄、斑块破裂等演变。
[Abstract]:Objective: to investigate the role of CD40-CD40L signal in the pathogenesis and development of human coronary atherosclerosis (as), the changes of lesion structure and the expression of CD40LMP-9 in human coronary atherosclerosis (as) were observed in this study. Methods: 60 cases of coronary artery as experimental group were divided into four groups according to pathological stages: lipid striated stage (n = 5), fibrous plaque (n = 19), atherosclerotic plaque (n = 18), secondary change (n = 18) and control group (n = 12). The tissue structure of coronary artery in both groups was observed by routine HE staining and the expression of CD40LmMMP-9 protein was detected by immunohistochemical SP two-step method with image analysis software. Reverse transcription-real-time fluorescence quantitative PCR (Reverse transcription-quantitative real time polymerase chain reaction (RT-PCR) was used to detect the expression of MMP-9 mRNA, and the relationship between the expression of MMP-9 and the structural changes of coronary atherosclerosis (as) was analyzed. The result is 1: 1. In the control group, the wall of coronary artery was thin, the thickness of each layer of intima, media and adventitia was uniform, and the intima was smooth and intact. The experimental group showed different changes from lipid-striated stage to secondary lesion stage. The lesion resulted in different degree of thickening of the coronary artery wall and stenosis of the lumen, calcification, hemorrhage, thrombosis and so on in some of the lesions, but only foam cell proliferation was observed in the light microscope. The thickness of vascular wall and lumen had no obvious change, and the severe cases had obvious atheronecrosis, even plaque hemorrhage or thrombosis, the wall of blood vessel was thickened significantly, and the lumen was obviously narrow. 2. The expression of CD40L was not found in the vascular wall of the control group, but the expression of CD40L was significantly increased in the experimental group, and the positive expression was mainly located in the shoulder region and the basal base of the as lesion. The mean optical density of CD40L in foam cells and inflammatory cells around the fibrous cap and necrotic foci was significantly higher in the experimental group than in the control group (P0.05), and the level of CD40L in each stage of the experimental group was significantly different (P0.05). In the control group, a small amount of MMP-9 protein was expressed in the cytoplasm of the medial smooth muscle cells and intimal foam cells, and in the coronary atherosclerotic plaques, the MMP-9 protein was expressed in the foam cells and inflammatory cells in the coronary atherosclerotic plaques. The fibrous cap area and the necrotic foci were obvious, and a few of them were expressed in the medial smooth muscle cells. The mean optical density showed that the level of MMP-9 in the experimental group was significantly higher than that in the control group (P0.05), and there was a significant difference in the expression of MMP-9 in different lesion stages (P0.05). 4. RT-qPCR method was used to detect the level of CD40LmMP-9 mRNA in coronary artery tissue. The results showed that the level of CD40L in the experimental group was higher than that in the control group (P0.05), and the level of MMP-9 mRNA in the experimental group was higher than that in the control group (P0.05). There was no significant difference between the experimental group and the control group (P0.05). The expression of protein CD40L was positively correlated with the expression of MMP-9, and they were positively correlated with the thickness of the thickest intima, the thickness of necrotic foci and the degree of vascular stenosis (P0.05), and had a negative correlation with the thickness fraction of the fibrous cap. Conclusion 1. The expression of CD40L in as lesions of human coronary artery was significantly increased. 2. 2. High expression of MMP-9 was found in as lesions of human coronary artery, and was positively correlated with CD40L expression, and was related to structural changes of lesions. 3. In human coronary atherosclerosis, CD40L may promote the expression of MMP-9, accelerate the degradation of extracellular matrix, and eventually lead to the formation of fibrous cap thinning and plaque rupture.
【学位授予单位】：贵州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.4

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