Neural Regeneration Research ›› 2016, Vol. 11 ›› Issue (12): 1981-1989.doi: 10.4103/1673-5374.197142

Previous Articles     Next Articles

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

Bo-jing Yan, Zhi-zhong Wu, Wei-hua Chong, Gen-lin Li*   

  1. Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Sciences Key Lab, Beijing, China
  • Received:2016-11-03 Online:2016-12-31 Published:2016-12-31
  • Contact: Gen-lin Li, M.D., ligenlin2018@163.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81271046; the Joint Program of Beijing Municipal Natural Science Foundation (category B) and Beijing Educational Committee (key project), No. KZ201510025025.

Abstract: 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 efcient treatment for fundus disease, an eukaryotic expression plasmid was generated and used to transfect human 293T cells to assess the expression and bioactivity of BDNF on acute retinal pigment epithelial-19 (ARPE-19) cells, a human retinal epithelial cell line. Afer 96 hours of co-culture in a Transwell chamber, ARPE- 19 cells exposed to BDNF secreted by 293T 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, 293T cells transfected with the BDNF gene steadily secreted the protein. Te powerful anti-apoptotic function of this BDNF may be useful for the treatment of retinitis pigmentosa and other retinal degenerative diseases.

Key words: nerve regeneration, neurodegenerative disease, brain-derived neurotrophic factor, retinitis pigmentosa, retina, retinal pigment pithelium, biosynthesis, transfection, plasmids, green ?uorescent protein, apoptosis, cell survival, neural regeneration