本文關(guān)鍵詞： 視網(wǎng)膜血管內(nèi)皮細(xì)胞 間充質(zhì)細(xì)胞 發(fā)育 轉(zhuǎn)型　出處：《青島大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討鼠視網(wǎng)膜血管正常發(fā)育過程中血管內(nèi)皮細(xì)胞轉(zhuǎn)型的具體機(jī)制。檢測細(xì)胞間CD31、VE-cadherin,間充質(zhì)樣細(xì)胞標(biāo)志蛋白平滑肌樣肌動(dòng)蛋白(a-SMA)、鈣結(jié)合樣蛋白(FSP-1)及細(xì)胞轉(zhuǎn)型調(diào)控基因Twist在鼠視網(wǎng)膜血管成熟過程中表達(dá)變化情況。 方法：對照實(shí)驗(yàn)研究將新生C57BL/6J小鼠40只,在生后第1、5、8、17天時(shí)各處死小鼠10只,應(yīng)用熒光素鈉灌注視網(wǎng)膜鋪片方法觀察視網(wǎng)膜血管發(fā)育的形態(tài)變化,應(yīng)用視網(wǎng)膜鋪片免疫熒光技術(shù)觀察鼠視網(wǎng)膜發(fā)育過程中血小板-內(nèi)皮細(xì)胞粘附分子(PECAM1/CD31)、平滑肌肌動(dòng)蛋白(smooth muscle actin, a-SMA)、成纖維細(xì)胞特異性蛋白-1(Fibroblast specific protein, Fsp-1)及Twist表達(dá)變化情況；提取全視網(wǎng)膜RNA,應(yīng)用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)檢測CD31、a-SMA、FSP-1及Twist基因表達(dá)變化情況,采用ΔΔCt法計(jì)算RQ值,使用SPSS13.0統(tǒng)計(jì)學(xué)軟件進(jìn)行單因素方差分析,以P0.05作為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果：視網(wǎng)膜鋪片免疫熒光技術(shù)結(jié)果顯示,小鼠CD31在生后第5天時(shí)表達(dá)較高,第8天和17天保持較低的水平,VE-cadherin在出后呈逐漸上升的趨勢,a-SMA和FSP-1表達(dá)分別在第1天和第5天較高,但其它時(shí)間均為較低水平,Twist表達(dá)在第5天較高。RT-PCR檢測結(jié)果顯示各組CD31(F=27.813,P0.05)、a-SMA(F=19.171,P0.05)、FSP-1(F=508.791,P0.05)、Twist(F=27.229,P0.05)RQ值間比較有統(tǒng)計(jì)學(xué)意義。 結(jié)論：在小鼠視網(wǎng)膜血管正常發(fā)育過程中可能發(fā)生了間充質(zhì)內(nèi)皮轉(zhuǎn)型,Twist基因可能參與了這個(gè)轉(zhuǎn)型過程。
[Abstract]:Objective: to investigate the mechanism of vascular endothelial cell transformation during the normal development of rat retinal vessels, and to detect CD31 VE-cadherin, smooth muscle like actin (SMA), calcium binding protein (FSP-1) and cell transformation modulation in mesenchymal cells. Expression of Twist gene in rat retinal vascular maturation. Methods: 40 newborn C57BL / 6J mice and 10 dead mice were killed at 817 days after birth. The morphological changes of retinal vascular development were observed by perfusion of fluorescein sodium into retinal slices. The expression of platelet-endothelial cell adhesion molecule (PECAM1 / CD31), smooth muscle actin smooth muscle actinin, a-SMA, fibroblast-specific protein (Fsp-1) and Twist during the development of rat retina were observed by immunofluorescence technique. The expression of FSP-1 and Twist was detected by reverse transcriptase polymerase chain reaction (RT-PCR). The RQ value was calculated by 螖 Ct method. The single factor variance analysis was performed by SPSS13.0 software. The difference was statistically significant (P0.05). Results: immunofluorescence technique showed that the expression of CD31 was higher in mice on the 5th day after birth. The expression of VE-cadherin increased gradually on day 8 and day 17, and the expression of a-SMA and FSP-1 increased on day 1 and day 5, respectively. On the 5th day, however, the expression of Twist was at a lower level. The results of RT-PCR showed that there was a significant difference between the RQ values of TwistF27.229P0.05RQ of TwistF27.229P0.05in each group. The results showed that the expression of TwistF27.229P0.05RQ in each group was significantly higher than that in the control group (P < 0.05), but the expression of Twist was lower at the other time. The results showed that the expression of TwistF27.229P0.05RQ in each group was significantly higher than that in the control group (P < 0.05). Conclusion: Twist gene may be involved in the normal development of retinal vessels in mice.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R774.1

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