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慢病毒介导的VEGF小发夹RNA对人Tenon成纤维细胞增殖的抑制作用

Suppression of Human Tenon Fibroblast Cell Proliferation by Lentivirus-Mediated VEGF Small Hairpin RNA.

作者信息

Li Zhongqiu, Hua Wen, Li Xuedong, Wang Wei

机构信息

Department of Ophthalmology, Chaoyang Hospital, Capital Medical University, Beijing, China.

出版信息

J Ophthalmol. 2017;2017:7982051. doi: 10.1155/2017/7982051. Epub 2017 Jan 11.

Abstract

. The functions of vascular endothelial growth factor (VEGF) in scar formation after trabeculectomy were investigated in a human Tenon fibroblast cell line from glaucoma patients using lentivirus-mediated VEGF shRNA. . Human Tenon fibroblast (HTF) cells were isolated from scar tissue of glaucoma patients during secondary surgery. Lentivirus-VEGF-shRNA was constructed and transfected into HTF cells. Subsequently, VEGF mRNA and protein expression were analyzed using quantitative RT-PCR and western blotting, respectively, and the effects of VEGF knockdown were analyzed. The inhibition of HTF proliferation was monitored according to total cell numbers using ScanArray. . Both mRNA and protein levels of VEGF were reduced by lentivirus-mediated VEGF-shRNA, and proliferation of HTF cells was inhibited. . Primary cultures of human Tenon fibroblast (HTF) were established, and proliferation was decreased following inhibition of VEGF. VEGF may be a suitable therapeutic target for reducing scar tissue formation in glaucoma patients after filtration surgery.

摘要

利用慢病毒介导的血管内皮生长因子(VEGF)短发夹RNA,在来自青光眼患者的人Tenon成纤维细胞系中研究了VEGF在小梁切除术后瘢痕形成中的作用。从青光眼患者二次手术时的瘢痕组织中分离出人Tenon成纤维细胞(HTF)。构建慢病毒-VEGF-shRNA并转染到HTF细胞中。随后,分别使用定量逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法分析VEGF mRNA和蛋白质表达,并分析VEGF基因敲低的效果。使用ScanArray根据总细胞数监测HTF增殖的抑制情况。慢病毒介导的VEGF-shRNA降低了VEGF的mRNA和蛋白质水平,并抑制了HTF细胞的增殖。建立了人Tenon成纤维细胞(HTF)的原代培养物,抑制VEGF后其增殖减少。VEGF可能是减少青光眼患者滤过手术后瘢痕组织形成的合适治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/164f/5266818/7e21aad829c6/JOPH2017-7982051.001.jpg

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