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人热敏瞬时受体通道1基因转染对猫角膜内皮细胞P27蛋白表达的影响
2012年
目的:探讨人热敏瞬时受体通道1(TRP1)基因转染对猫角膜内皮细胞P27蛋白表达的影响,探讨热敏TRP1基因对角膜内皮细胞增殖周期的作用,为该基因应用于人角膜内皮细胞提供理论依据。方法:将20只健康足月幼猫随机分为实验组和对照组,每组10只,麻醉后取角膜内皮细胞进行培养,培养后实验组细胞通过脂质体介导的方法转染人热敏TRP1基因,对照组采用5.0μL PBS代替表达载体加入培养基。RT-PCR观察人热敏TRP1基因mRNA表达水平,Western blotting观察猫角膜内皮细胞内P27蛋白的表达。继续培养96h观察目的基因转染对细胞分裂再生活性的影响。结果:细胞培养、加热后钙离子流入明显增多;人热敏TRP1基因转染后RT-PCR结果显示。实验组条带密度显著高于对照组(t=2.21,P=0.037);Western blotting结果显示,实验组P27蛋白表达显著高于对照组,差异有统计学意义(t=2.53,P=0.021);实验组和对照组有丝分裂指数、G1期细胞比例差异均有统计学意义(χ2=3.26,χ2=4.18,P<0.05)。结论:人热敏TRP1基因转染可以增加猫角膜内皮细胞P27蛋白表达,抑制角膜内皮细胞增殖。
王莉李鹏杜兆江
关键词:基因转染角膜内皮细胞P27蛋白
Magnetic nanoparticles conjugated with “RPE cell-MCP-1 antibody-VEGF antibody” compounds for the targeted therapy of age-related macular degeneration: a hypothesis被引量:1
2017年
Age-related macular degeneration(AMD) is the leading cause of vision loss in the elderly throughout the world. Treatment of AMD utilizing retinal pigment epithelium(RPE) transplantation represents a promising therapy. However, simplex RPE transplantation can only replace the diseased RPE cells, but has no abilities to stop the development of AMD. It has been indicated that oxidization triggers the development of AMD by inducing the dysfunction and degeneration of RPE cells, which results in the upregulation of local monocyte chemotactic protein-1(MCP-1) expression. MCP-1 induces macrophage recruiment which triggers local inflammation. As a result, the expression of vascular endothelial growth factor(VEGF) is upregulated by MCP-1mediated inflammation and results in the formation of choroidal neovascularization(CNV). We accordingly propose a targeted therapy of AMD by subretinal transplanting the compound of RPE cell, MCP-1 antibody, and VEGF antibody and using a magnetic system to guide RPE cell compounds conjugated with superparamagnetic iron oxide nanoparticles(SPIONs). Furthermore, SPION-labelled RPE cells can be tracked and detected in vivo by non-invasive magnetic resonance imaging(MRI). This novel RPE cell transplantation methodology seems very promising to provide a new therapeutic approach for the treatment of AMD.
Zhao-Jiang DuPeng LiLi Wang
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