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Construction of a plasmid for human brain-derived neurotrophic factor and its effect on retinal pigment epithelial cell viability 被引量:2

Construction of a plasmid for human brain-derived neurotrophic factor and its effect on retinal pigment epithelial cell viability
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摘要 Several studies have investigated the protective functions of brain-derived neurotrophic factor(BDNF) in retinitis pigmentosa. However, a BDNF-based therapy for retinitis pigmentosa is not yet available. To develop an efficient treatment for fundus disease, an eukaryotic expression plasmid was generated and used to transfect human 293 T cells to assess the expression and bioactivity of BDNF on acute retinal pigment epithelial-19(ARPE-19) cells, a human retinal epithelial cell line. After 96 hours of co-culture in a Transwell chamber, ARPE-19 cells exposed to BDNF secreted by 293 T cells were more viable than ARPE-19 cells not exposed to secreted BDNF. Western blot assay showed that Bax levels were downregulated and that Bcl-2 levels were upregulated in human ARPE-19 cells exposed to BDNF. Furthermore, 293 T cells transfected with the BDNF gene steadily secreted the protein. The powerful anti-apoptotic function of this BDNF may be useful for the treatment of retinitis pigmentosa and other retinal degenerative diseases. Several studies have investigated the protective functions of brain-derived neurotrophic factor(BDNF) in retinitis pigmentosa. However, a BDNF-based therapy for retinitis pigmentosa is not yet available. To develop an efficient treatment for fundus disease, an eukaryotic expression plasmid was generated and used to transfect human 293 T cells to assess the expression and bioactivity of BDNF on acute retinal pigment epithelial-19(ARPE-19) cells, a human retinal epithelial cell line. After 96 hours of co-culture in a Transwell chamber, ARPE-19 cells exposed to BDNF secreted by 293 T cells were more viable than ARPE-19 cells not exposed to secreted BDNF. Western blot assay showed that Bax levels were downregulated and that Bcl-2 levels were upregulated in human ARPE-19 cells exposed to BDNF. Furthermore, 293 T cells transfected with the BDNF gene steadily secreted the protein. The powerful anti-apoptotic function of this BDNF may be useful for the treatment of retinitis pigmentosa and other retinal degenerative diseases.
出处 《Neural Regeneration Research》 SCIE CAS CSCD 2016年第12期1981-1989,共9页 中国神经再生研究(英文版)
基金 supported by the National Natural Science Foundation of China,No.81271046 the Joint Program of Beijing Municipal Natural Science Foundation(category B) Beijing Educational Committee(key project),No.KZ201510025025
关键词 nerve regeneration neurodegenerative disease brain-derived neurotrophic factor retinitis pigmentosa retina retinal pigment epithelium biosynthesis transfection plasmids green fluorescent protein apoptosis cell survival neural regeneration nerve regeneration neurodegenerative disease brain-derived neurotrophic factor retinitis pigmentosa retina retinal pigment epithelium biosynthesis transfection plasmids green fluorescent protein apoptosis cell survival neural regeneration
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