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 ...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.展开更多
Autophagy plays a vital role in maintaining the balance of normal physiological state of living cells.In this paper,a polarity-specific two-phot on fluorescent probe Lyso-NA based on naphthalimide was synthesized for ...Autophagy plays a vital role in maintaining the balance of normal physiological state of living cells.In this paper,a polarity-specific two-phot on fluorescent probe Lyso-NA based on naphthalimide was synthesized for the purpose of monitoring autophagy during biological research.The results of photophysical properties and theoretical calculation con firmed that different polarities of solvents mainly effected fluorescent intensities of probe.Fluoresce nt intensity,quantum yield and fluorescence lifetime of probe kept a good linear relationship with polarity respectively.In addition,due to its low toxicity and selective accumulation in lysosomes,Lyso-NA is suitable for detecting changes in lysosomal polarity of living cells.Compare with the imaging results of plasmid transfection,a better performed realtime long-term fluoresce nt visualization of autophagy in living cells was achieved.Probe Lyso-NA can work as an efficient and cost effective imaging tool for visualizing autophagy in living cells.展开更多
基金supported by the National Natural Science Foundation of China,No.81271046the Joint Program of Beijing Municipal Natural Science Foundation(category B)Beijing Educational Committee(key project),No.KZ201510025025
文摘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.
基金This work was supported by the National Natural Science Foundation of China(Nos.22077001,21778001 and 21672001)the Natural Science Foundation of Anhui Province(No.2008085J08)+2 种基金the Natural Science Foundation of Education Department of Anhui Province(No.KJ2019A0010)Joint Open Fund of Jiangsu Collaborative Innovation Center for Ecological Building Material and Environmental Protection Equipments,Key Laboratory for Advanced Technology in Environmental Protection of Jiangsu Province(No.JH201803)the Open fund for Discipline Construction in Institute of Physical Science and In formation Technology of Anhui University.
文摘Autophagy plays a vital role in maintaining the balance of normal physiological state of living cells.In this paper,a polarity-specific two-phot on fluorescent probe Lyso-NA based on naphthalimide was synthesized for the purpose of monitoring autophagy during biological research.The results of photophysical properties and theoretical calculation con firmed that different polarities of solvents mainly effected fluorescent intensities of probe.Fluoresce nt intensity,quantum yield and fluorescence lifetime of probe kept a good linear relationship with polarity respectively.In addition,due to its low toxicity and selective accumulation in lysosomes,Lyso-NA is suitable for detecting changes in lysosomal polarity of living cells.Compare with the imaging results of plasmid transfection,a better performed realtime long-term fluoresce nt visualization of autophagy in living cells was achieved.Probe Lyso-NA can work as an efficient and cost effective imaging tool for visualizing autophagy in living cells.
