RESUMEN
The present study aimed to investigate the effect of arsenic trioxide (ATO) on the proliferation of retinal pigment epithelium (RPE) and its mechanism. RPE cells were cultivated with 0.5-11 µmol/L ATO for 24, 48, and 72 h and their survival and growth were measured by MTT assay. The expression of p27 and proliferating cell nuclear antigen (PCNA) in RPE cells was detected using cell immunofluorescence and western blotting. Dose-dependency was evident in both the experimental and control groups. The 50% inhibitory concentration was obtained at a concentration of 6 mol/L with cells treated for 3 days. The optimum concentration of ATO was 6 µmol/L based on the result of MTT. After the third day of ATO treatment, the number of cells was significantly lower in the experimental group compared with the control group. The expression of extracellular matrix (ECM) components decreased relative to the control group. The expression of p27 and PCNA declined gradually in cells treated for 72 h at 6 µmol/L ATO compared with the control group. The difference between the experimental and control groups was significant (P=0.005). ATO has the ability to inhibit the growth and proliferation of RPE cells by regulating the expression of the ECM components' p27 and PCNA, in a time- and dose-dependent manner. Thus, ATO may lead to an innovative method for the treatment of proliferative retinopathy.
RESUMEN
The recycling of neurotransmitters is essential for sustained synaptic transmission. In Drosophila, histamine recycling is required for visual synaptic transmission. Synaptic histamine is rapidly taken up by laminar glia, and is converted to carcinine. After delivered back to photoreceptors, carcinine is hydrolyzed to release histamine and ß-alanine. This histamine is repackaged into synaptic vesicles, but it is unclear how the ß-alanine is returned to the laminar glial cells. Here, we identified a new ß-alanine transporter, which we named BalaT (Beta-alanine Transporter). Null balat mutants exhibited lower levels of ß-alanine, as well as less ß-alanine accumulation in the retina. Moreover, BalaT is expressed and required in retinal pigment cells for maintaining visual synaptic transmission and phototaxis behavior. These results provide the first genetic evidence that retinal pigment cells play a critical role in visual neurotransmission, and suggest that a BalaT-dependent ß-alanine trafficking pathway is required for histamine homeostasis and visual neurotransmission.
Asunto(s)
Drosophila/fisiología , Proteínas de Transporte de Membrana/metabolismo , Epitelio Pigmentado de la Retina/metabolismo , Transmisión Sináptica , beta-Alanina/metabolismo , Animales , Técnicas de Inactivación de Genes , Histamina/metabolismo , Proteínas de Transporte de Membrana/genética , Neuroglía/metabolismoRESUMEN
To study the cellular uptake mechanism of multifunctional organic-inorganic hybrid nanoparticles and nanosheets, new chitosan-glutathione-valine-valine-layered double hydroxide (CG-VV-LDH) nanosheets with active targeting to peptide transporter-1 (PepT-1) were prepared, characterized and further compared with CG-VV-LDH nanoparticles. Both organic-inorganic hybrid nanoparticles and nanosheets showed a sustained release in vitro and prolonged precorneal retention time in vivo, but CG-VV-LDH nanoparticles showed superior permeability in the isolated cornea of rabbits than CG-VV-LDH nanosheets. Furthermore, results of cellular uptake on human corneal epithelial primary cells (HCEpiC) and retinal pigment epithelial (ARPE-19) cells indicated that both clathrin-mediated endocytosis and active transport of PepT-1 are involved in the internalization of CG-VV-LDH nanoparticles and CG-VV-LDH nanosheets. In summary, the CG-VV-LDH nanoparticle may be a promising carrier as a topical ocular drug delivery system for the treatment of ocular diseases of mid-posterior segments, while the CG-VV-LDH nanosheet may be suitable for the treatment of ocular surface diseases.