RESUMEN
OBJECTIVE: With the rabbit iris pigment epithelial cells (IPECs), which were containing the NT4-NAP fusion gene, taken as the substituting secreting cells producing the neuropeptide NAP, the effect of neuropeptide NAP on the growth status of retinal neuroepithelial cells of rabbit was examined and explored. METHODS: The iris pigment epithelial cells and retinal neuroepithelial cells of rabbit were cultured; rAAV-GFP and rAAV-NAP (containing NT4-NAP fusion gene) were constructed; the rabbit IPECs were infected with rAAV-GFP and rAAV-NAP; the infections of viruses were detected by GFP fluorescence expression; the supernatant of culture from rabbit IPECs with NAP was collected and added into the culture medium of rabbit retinal neuroepithelial cells, and the growth state of retinal neuroepithelial cell was observed. RESULTS: Compared to control cells, the rabbit IPECs could express the GFP fluorescence. The rabbit retinal neuroepithelial cells with NAP supernatant could survive more, and the survival cells showed longer and stronger axons. On 14's day of cell culture, the mean axon length of NAP group was (14. 6+/-1. 1) microm, while that of control group was only (3. 1+/-0. 6) mircom. Obviously, a significant difference existed between two groups (P<0. 05). CONCLUSION: The rabbit IPECs with NT4-NAP fusion gene can secrete NAP, and the NAP can promise the growth of rabbit retinal
Asunto(s)
Células Neuroepiteliales/citología , Células Neuroepiteliales/efectos de los fármacos , Neuropéptidos/farmacología , Oligopéptidos/farmacología , Retina/citología , Animales , Línea Celular , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Dependovirus/genética , Dependovirus/fisiología , Femenino , Iris/citología , Masculino , Células Neuroepiteliales/metabolismo , Neuropéptidos/genética , Oligopéptidos/genética , Conejos , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/farmacología , Epitelio Pigmentado de la Retina/citología , Carga ViralRESUMEN
OBJECTIVE: To investigate the effect of infection with adeno-associated virus (AAV) vector containing NT4-NAP fusion gene on photoprotection of rabbit iris pigment epithelium cells (IPECs). METHODS: rAAV-GFP and rAAV-NAP (containing NT4-NAP fusion gene) were constructed; rabbit IPECs were cultured and infected with rAAV-GFP and rAAV-NAP; transfection of viruses was detected by GFP fluorescence expression; NAP protect rabbit iris pigment epithelium from light stimulation was evaluated by MTT and flow cytometry. RESULTS: Rabbit IPECs expressed the GFP fluorescence; comparing with control cells, IPECs transfected with rAAV-NAP remained normal proliferation and showed lower apoptosis percentage after ultra-violet stimulation. CONCLUSION: rAAV-NAP constructed in the study can infect rabbit IPECs, and NAP may protect rabbit IPECs from ultra-violet damage.