Correlation Study Of Activated Astrocytes Of Optic Nerve Head In The Rat And Subtypes Of PGE2/EP Receptor

Objective: We explore the expression features and effect on astrocytes of EP receptor after astrocytes of optic nerve head activating.Methods: Primary culture of astrocytes of optic nerve head: we took off the eyeballs of 3-month-old SD rats,then extracted the optic nerve head(ONH)under surgical microscope.ONH was cut into 2 or 3 pieces,and put into ordinary culture medium DMEM/F12 which contained 10% fetal calf serum.We got rid of the tissue after the cells climbed out from it,and put the cells into astrocytes selective medium.And GFAP cells was used to identify after subculture purification and segregation of astrocytes twice.Western-blotting was used to detect the expression of EP receptor of SD rat,C57 mice and healthy human.Imunofluorescence was used to detect the expression localization of activated astrocytes EP receptor in non-administered group and given epidermal growth factor(EGF/NMDA)group respectively.Westernblotting was used to test the expression of COX-2,EP receptors and GFAP after EGF(20ng/ml)and the different density of NMDA activated astrocytes.Results: GFAP was expressed in about 95 percent of purified astrocytes after3-week cultivation.The results of Western-blotting showed that EP receptor of SD rat,C57 mice and healthy human all expressed.The results of imunofluorescence showed EP1 express in the cell membrane,EP2 and EP4 in cell nucleus,and EP3 in the cytoplasm,although given EGF the results also.EGF could activate astrocytes and changed its morphology,and increased GFAP expression(P < 0.05).We found COX-2 was induced formation,and the expression of four receptors of PEG2 was increased,and there was a statistical significance(P<0.05).Conclusion: the expression of EP receptor of PGE2 was increased after EGF activated astrocytes,and EP receptor participated in astrocytes activation reaction.