鼠视网膜血管发育中的内皮细胞转型机制

发布时间：2018-02-24 09:37

本文关键词： 视网膜血管内皮细胞间充质细胞发育转型　出处：《青岛大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的：探讨鼠视网膜血管正常发育过程中血管内皮细胞转型的具体机制。检测细胞间CD31、VE-cadherin,间充质样细胞标志蛋白平滑肌样肌动蛋白(a-SMA)、钙结合样蛋白(FSP-1)及细胞转型调控基因Twist在鼠视网膜血管成熟过程中表达变化情况。方法：对照实验研究将新生C57BL/6J小鼠40只,在生后第1、5、8、17天时各处死小鼠10只,应用荧光素钠灌注视网膜铺片方法观察视网膜血管发育的形态变化,应用视网膜铺片免疫荧光技术观察鼠视网膜发育过程中血小板-内皮细胞粘附分子(PECAM1/CD31)、平滑肌肌动蛋白(smooth muscle actin, a-SMA)、成纤维细胞特异性蛋白-1(Fibroblast specific protein, Fsp-1)及Twist表达变化情况；提取全视网膜RNA,应用逆转录聚合酶链反应检测CD31、a-SMA、FSP-1及Twist基因表达变化情况,采用ΔΔCt法计算RQ值,使用SPSS13.0统计学软件进行单因素方差分析,以P0.05作为差异有统计学意义。结果：视网膜铺片免疫荧光技术结果显示,小鼠CD31在生后第5天时表达较高,第8天和17天保持较低的水平,VE-cadherin在出后呈逐渐上升的趋势,a-SMA和FSP-1表达分别在第1天和第5天较高,但其它时间均为较低水平,Twist表达在第5天较高。RT-PCR检测结果显示各组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值间比较有统计学意义。结论：在小鼠视网膜血管正常发育过程中可能发生了间充质内皮转型,Twist基因可能参与了这个转型过程。
[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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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