纤维蛋白原对内皮祖细胞血管生成功能的影响

发布时间：2018-04-16 14:34

本文选题：内皮祖细胞 + 纤维蛋白原　；参考：《首都医科大学》2017年硕士论文

【摘要】：背景:内皮祖细胞(EPCs)与纤维蛋白原可参与损伤血管的修复与新生血管形成。在内皮损伤后EPCs可由骨髓动员并归巢于损伤部位参与血管修复及新生血管形成。纤维蛋白原可通过促进ECs的迁移与增殖功能而参与肿瘤组织中新生血管的形成。研究显示长期接触纤维蛋白原可不同程度增加肺动脉内皮细胞(PAECs)与肺动脉平滑肌细胞(PASMCs)内基础钙(Ca2+)浓度,影响细胞的迁移与增殖功能。目的:将大鼠骨髓EPCs暴露于纤维蛋白原环境中培养后,对其血管生成功能进行检测,初步探讨纤维蛋白原对EPCs血管生成功能影响。方法:大鼠骨髓EPCs于体外培养9天后,将其接种于4μg/ml与40μg/ml两种浓度的纤维蛋白原包板的玻片上,孵育72小时后,进行血管生成实验,并使用Image J对血管网状结构长度、小管数、分支点数进行定量化分析。结果:EPCs体外培养的形态学变化:细胞培养1天后可见细胞贴壁,大多数细胞为圆形,4天后部分贴壁细胞为梭形或多边形,可见部分细胞排列呈线状,6天后贴壁梭形或多边形细胞增加,且呈优势生长,8天后贴壁梭形或多边形细胞呈优势生长,呈铺路石样紧密排列。EPCs化学免疫荧光染色鉴定:Dil-ac-LDL和FITC-UEA-1双阳性细胞占85.726%±2.568%(n=3)。血管生成功能检测:细胞接种于由Matrigel包被的24孔板孵育5小时后可见明显血管网状结构形成,血管网状结构长度为9.782±0.666mm,小管数为7.433±0.286个,分支点数为8.533±1.347个(n=3)对照组:血管网状结构长度为9.975±0.563mm,小管数为8.967±0.260个,分支点数为11.800±0.436个(n=3);4μg/ml纤维蛋白原组血管网状结构长度为12.604±1.354mm(P=0.231),小管数为10.767±0.649个(P=0.064),分支点数为14.400±1.179个(P=0.071)(n=3),较对照组无明显差异。40μg/ml纤维蛋白原组血管网状结构长度为25.764±1.920mm(P=00.000),小管数为16.133±0.677个(P=0.001),分支点数为21.833±0.726个(P=0.001)(n=3),较对照组有明显增加。4μg/ml组与40μg/ml组相比:血管网状结构(P=0.001)、小管数(P=0.002)及分支点数(P=0.001)均有明显增加,有统计学差异。结论:纤维蛋白原可促进EPCs的血管生成功能,且呈浓度依赖性,但其机制仍需进一步探讨。
[Abstract]:Background: endothelial progenitor cells (EPCs) and fibrinogen are involved in the repair and angiogenesis of injured blood vessels.After endothelial injury, EPCs can be mobilized by bone marrow and homing to the injured site to participate in vascular repair and angiogenesis.Fibrinogen can play a role in angiogenesis by promoting the migration and proliferation of ECs.The results showed that long-term exposure to fibrinogen could increase the basic Ca ~ (2 +) concentration in pulmonary artery endothelial cells (PAECs) and pulmonary artery smooth muscle cells (PASMCs), and affect the migration and proliferation of the cells.Aim: to detect the angiogenesis function of rat bone marrow EPCs exposed to fibrinogen, and to explore the effect of fibrinogen on EPCs angiogenesis.Methods: after cultured in vitro for 9 days, rat bone marrow EPCs was inoculated on the glass of fibrinogen coated with 4 渭 g/ml and 40 渭 g/ml, incubated for 72 hours, the angiogenesis experiment was carried out, and Image J was used to measure the length of vascular reticular structure and the number of tubules.The number of branches is analyzed quantitatively.Results the morphological changes of EPCs cultured in vitro were as follows: after 1 day of cell culture, adherent cells could be seen, and most of the cells were fusiform or polygonal after 4 days.It can be seen that some of the cells arranged in the form of linear or polygonal cells increased after 6 days, and showed dominant growth after 8 days of adherent fusiform or polygonal cells.The proportion of double positive cells of FITC-UEA-1 and Dil-ac-LDL was 85.726% 卤2.568%.Angiogenesis function test: after inoculation with Matrigel coated 24 hole plate for 5 hours, the formation of vascular reticular structure was obvious. The length of vascular reticular structure was 9.782 卤0.666 mm, the number of tubules was 7.433 卤0.286 mm, and the number of tubules was 7.433 卤0.286 mm.The number of branches was 8.533 卤1.347) in the control group, the length of vascular reticular structure was 9.975 卤0.563mm, and the number of tubules was 8.967 卤0.260.The number of branches in the fibrinogen group was 12.604 卤1.354mm, the number of tubules was 10.767 卤0.64mm, and the number of branches was 14.400 卤1.179 g/ml. There was no significant difference. 40 渭 g/ml fibrinogen group was 25.764 卤1.920mmP00.000, and the number of tubules was 16.133 卤0.677.Compared with the 40 渭 g/ml group, there were significant increases in the number of branches (21.833 卤0. 726) and P0. 001 (P0. 001) in the control group. Compared with the 40 渭 g/ml group, there was a significant increase in the number of vascular reticular structure (P0. 001), the number of tubules (P0. 002) and the number of branches (P0. 001).There is a statistical difference.Conclusion: fibrinogen can promote angiogenesis of EPCs in a concentration-dependent manner, but its mechanism needs further study.
【学位授予单位】：首都医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563

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