Mu Qing-Jie, Zhao Yue-Hua, Cheng Dan-Dan, Wang Hai-Yu, Chen Lan-Fen, Zhao Yan-Song, Wang Xiao-Li
Clinical Institute, Weifang Medical University, Weifang, Shandong 261053, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2017 Nov;19(11):1202-1207. doi: 10.7499/j.issn.1008-8830.2017.11.015.
To explore the effects of rat bone mesenchymal stem cell (BMSC) transplantation on retinal neovascularization, and to observe the changes of hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factors (VEGF) in rats with oxygen-induced retinopathy (OIR).
Seventy-two seven-day-old Sprague-Dawley rats were randomly divided into three groups: normal control (CON), model (OIR) and BMSC transplantation. In the BMSC transplantation group, BMSCs were transplanted 5 days after oxygen conditioning. The phosphate buffered saline of the same volume was injected in the CON and OIR groups. The OIR model was prerpared according to the classic hyperoxygen method. At seven days after transplantation, retinal neovascularization was examined by retinal flat-mount staining and hematoxylin eosin (HE) staining. The expression of HIF-1α and VEGF proteins was examined by immunohistochemistry staining and Western blot analysis.
The retinal flat-mount staining results showed that the vessels were well organized in the CON group, but the vessels were irregularly organized, and lots of nonperfusion areas were observed in the OIR group. The large vessels were a bit circuitous, the retinal vessels were relatively organized, and less nonperfusion areas were noted in the BMSC transplantation group. The HE staining results showed that many neovessels and preretinal neovascular (pre-RNC) cells were observed on the internal limiting membrane in the OIR group. There were less pre-RNC cells in the BMSC transplantation group compared with the OIR group (P<0.01). The immunohistochemistry analysis showed that more HIF-1α and VEGF cells were observed in the OIR group compared with the CON group, and less HIF-1α and VEGF cells were observed in the BMSC transplantation group compared with OIR group (P<0.05). The Western blot analysis showed the expression of HIF-1α and VEGF proteins in the OIR group was significantly higher than that in the CON group. The expression of HIF-1α and VEGF proteins in the BMSC transplantation group was lower than that in the OIR group (P<0.01).
BMSC transplantation therapy could alleviate retinal neovascularization in OIR rats, and its mechanisms might be associated with the inhibition of the expression of HIF-1α and VEGF proteins.
探讨大鼠骨髓间充质干细胞（BMSC）移植对视网膜新生血管形成的影响，并观察氧诱导视网膜病变（OIR）大鼠中缺氧诱导因子-1α（HIF-1α）和血管内皮生长因子（VEGF）的变化。
将72只7日龄的Sprague-Dawley大鼠随机分为三组：正常对照组（CON）、模型组（OIR）和BMSC移植组。在BMSC移植组中，氧诱导后5天进行BMSC移植。CON组和OIR组注射相同体积的磷酸盐缓冲盐水。按照经典的高氧方法制备OIR模型。移植后7天，通过视网膜铺片染色和苏木精-伊红（HE）染色检查视网膜新生血管形成情况。通过免疫组织化学染色和蛋白质免疫印迹分析检测HIF-1α和VEGF蛋白的表达。
视网膜铺片染色结果显示，CON组血管排列整齐，而OIR组血管排列不规则，观察到许多无灌注区。BMSC移植组大血管稍迂曲，视网膜血管相对排列整齐，无灌注区较少。HE染色结果显示，OIR组在内界膜上观察到许多新生血管和视网膜前新生血管（pre-RNC）细胞。与OIR组相比，BMSC移植组的pre-RNC细胞较少（P<0.01）。免疫组织化学分析显示，与CON组相比，OIR组观察到更多的HIF-1α和VEGF细胞，与OIR组相比，BMSC移植组观察到的HIF-1α和VEGF细胞较少（P<0.05）。蛋白质免疫印迹分析显示，OIR组中HIF-1α和VEGF蛋白的表达明显高于CON组。BMSC移植组中HIF-1α和VEGF蛋白的表达低于OIR组（P<0.01）。
BMSC移植治疗可减轻OIR大鼠的视网膜新生血管形成，其机制可能与抑制HIF-1α和VEGF蛋白的表达有关。
