發(fā)布時(shí)間：2018-04-16 14:34

  本文選題：內(nèi)皮祖細(xì)胞 + 纖維蛋白原��； 參考：《首都醫(yī)科大學(xué)》2017年碩士論文

【摘要】：背景:內(nèi)皮祖細(xì)胞(EPCs)與纖維蛋白原可參與損傷血管的修復(fù)與新生血管形成。在內(nèi)皮損傷后EPCs可由骨髓動(dòng)員并歸巢于損傷部位參與血管修復(fù)及新生血管形成。纖維蛋白原可通過促進(jìn)ECs的遷移與增殖功能而參與腫瘤組織中新生血管的形成。研究顯示長(zhǎng)期接觸纖維蛋白原可不同程度增加肺動(dòng)脈內(nèi)皮細(xì)胞(PAECs)與肺動(dòng)脈平滑肌細(xì)胞(PASMCs)內(nèi)基礎(chǔ)鈣(Ca2+)濃度,影響細(xì)胞的遷移與增殖功能。目的:將大鼠骨髓EPCs暴露于纖維蛋白原環(huán)境中培養(yǎng)后,對(duì)其血管生成功能進(jìn)行檢測(cè),初步探討纖維蛋白原對(duì)EPCs血管生成功能影響。方法:大鼠骨髓EPCs于體外培養(yǎng)9天后,將其接種于4μg/ml與40μg/ml兩種濃度的纖維蛋白原包板的玻片上,孵育72小時(shí)后,進(jìn)行血管生成實(shí)驗(yàn),并使用Image J對(duì)血管網(wǎng)狀結(jié)構(gòu)長(zhǎng)度、小管數(shù)、分支點(diǎn)數(shù)進(jìn)行定量化分析。結(jié)果:EPCs體外培養(yǎng)的形態(tài)學(xué)變化:細(xì)胞培養(yǎng)1天后可見細(xì)胞貼壁,大多數(shù)細(xì)胞為圓形,4天后部分貼壁細(xì)胞為梭形或多邊形,可見部分細(xì)胞排列呈線狀,6天后貼壁梭形或多邊形細(xì)胞增加,且呈優(yōu)勢(shì)生長(zhǎng),8天后貼壁梭形或多邊形細(xì)胞呈優(yōu)勢(shì)生長(zhǎng),呈鋪路石樣緊密排列。EPCs化學(xué)免疫熒光染色鑒定:Dil-ac-LDL和FITC-UEA-1雙陽性細(xì)胞占85.726%±2.568%(n=3)。血管生成功能檢測(cè):細(xì)胞接種于由Matrigel包被的24孔板孵育5小時(shí)后可見明顯血管網(wǎng)狀結(jié)構(gòu)形成,血管網(wǎng)狀結(jié)構(gòu)長(zhǎng)度為9.782±0.666mm,小管數(shù)為7.433±0.286個(gè),分支點(diǎn)數(shù)為8.533±1.347個(gè)(n=3)對(duì)照組:血管網(wǎng)狀結(jié)構(gòu)長(zhǎng)度為9.975±0.563mm,小管數(shù)為8.967±0.260個(gè),分支點(diǎn)數(shù)為11.800±0.436個(gè)(n=3);4μg/ml纖維蛋白原組血管網(wǎng)狀結(jié)構(gòu)長(zhǎng)度為12.604±1.354mm(P=0.231),小管數(shù)為10.767±0.649個(gè)(P=0.064),分支點(diǎn)數(shù)為14.400±1.179個(gè)(P=0.071)(n=3),較對(duì)照組無明顯差異。40μg/ml纖維蛋白原組血管網(wǎng)狀結(jié)構(gòu)長(zhǎng)度為25.764±1.920mm(P=00.000),小管數(shù)為16.133±0.677個(gè)(P=0.001),分支點(diǎn)數(shù)為21.833±0.726個(gè)(P=0.001)(n=3),較對(duì)照組有明顯增加。4μg/ml組與40μg/ml組相比:血管網(wǎng)狀結(jié)構(gòu)(P=0.001)、小管數(shù)(P=0.002)及分支點(diǎn)數(shù)(P=0.001)均有明顯增加,有統(tǒng)計(jì)學(xué)差異。結(jié)論:纖維蛋白原可促進(jìn)EPCs的血管生成功能,且呈濃度依賴性,但其機(jī)制仍需進(jìn)一步探討。
[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% 鹵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 鹵0.666 mm, the number of tubules was 7.433 鹵0.286 mm, and the number of tubules was 7.433 鹵0.286 mm.The number of branches was 8.533 鹵1.347) in the control group, the length of vascular reticular structure was 9.975 鹵0.563mm, and the number of tubules was 8.967 鹵0.260.The number of branches in the fibrinogen group was 12.604 鹵1.354mm, the number of tubules was 10.767 鹵0.64mm, and the number of branches was 14.400 鹵1.179 g/ml. There was no significant difference. 40 渭 g/ml fibrinogen group was 25.764 鹵1.920mmP00.000, and the number of tubules was 16.133 鹵0.677.Compared with the 40 渭 g/ml group, there were significant increases in the number of branches (21.833 鹵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.
【學(xué)位授予單位】：首都醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R563

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