间充质干细胞对视网膜新生血管的影响和机制研究

Retinal neovascularization(RNV) is a common cuase of blindness all over the world, the treatment of which mainly depends on intravitreal injection of anti-VEGF drugs. Because of drug-nonresponse and the retinal atrophy, it is extremely urgent for us to discover new therapies for RNV. According to our previous studies, mesenchymal stem cells (MSCs) could differentiate into retinal cells, inhibit retinal apoptosis by expressing neurotrophic factors, and regulate retinal angiogenesis through microenvironment. In this proposal, we first investigate the effects of gene-modified MSCs on retinal vascular endothelial cells cultured in vitro via a coculture system. After genetic modification, MSCs are induced to express more antiangiogenic factors or less angiogenic factors. Raman spectrum, RT-PCR and transmission electron microscope, ect, will be conducted to analyze the effects of MSCs on retinal vascular endothelial cells. An animal model of hypoxia-induced retinal neovascularization will be treated using genetically modified MSCs in vivo. The migration, location and differentiation of MSCs is to be detected by laser scanning confocal microscopy. The site of neurotrophic factors and antiangiogenic factors expression will be identified by in situ hybridization, and quantitatively measured by RT-PCR and Western Blot . The interaction between MCSs and retinal vascular endothelial cells will be evaluated through FFA, OCT and ERG, etc, eventually the mechanism would be demonstrated. This proposal is promising in providing a new way for treating RNV.

视网膜新生血管性疾病是世界性致盲眼病，目前治疗主要为玻璃体腔注射抗VEGF药，而部分病人长期治疗无应答甚至产生视网膜萎缩，故研究RNV干预机制迫在眉睫。我们研究发现间充质干细胞(MSCs)可分化为视网膜细胞，分泌神经营养因子抑制其凋亡，还可调节视网膜血管生成的微环境。本研究拟通过①基因修饰MSCs，使血管生成抑制因子过表达/新生血管因子下调，MSCs与缺氧诱导的视网膜血管内皮细胞体外共培养，利用拉曼光谱、RT-PCR和透射电镜等检测MSCs对内皮细胞的影响。②将MSCs植入氧诱导视网膜新生血管小鼠的视网膜下，激光共聚焦测MSCs的迁移、定位及分化，RT-PCR和Western Blotting、原位杂交对神经营养因子和血管因子进行定量、定位。③FFA、OCT和ERG等评估MSCs视网膜血管的相互作用，研究MSCs对RNV的影响机制。有望为干预RNV提供新途径。

视网膜新生血管性疾病是世界性致盲眼病，目前治疗主要为玻璃体腔注射抗VEGF药，而部分病人长期治疗无应答甚至产生视网膜萎缩，故研究RNV干预机制迫在眉睫。我们研究发现间充质干细胞(MSCs)可分化为视网膜细胞，分泌神经营养因子抑制其凋亡，还可调节视网膜血管生成的微环境。本研究通过①基因修饰MSCs，使血管生成抑制因子过表达/新生血管因子下调，MSCs与缺氧诱导的视网膜血管内皮细胞体外共培养，利用拉曼光谱、RT-PCR和透射电镜等检测MSCs对内皮细胞的影响。②将MSCs植入氧诱导视网膜新生血管小鼠的视网膜下，激光共聚焦测MSCs的迁移、定位及分化，RT-PCR和Western Blotting、原位杂交对神经营养因子和血管因子进行定量、定位。③FFA、OCT和ERG等评估MSCs视网膜血管的相互作用，研究MSCs对RNV的影响机制。本研究成果有望为干预RNV提供新途径。