Expression Of GLAST And Unfolded Protein Reaction Related Factors XBP1、CHOP In Retinal Müller Cell By Hypoxia

Glaucoma,age-related macular degeneration,retinal pigment degeneration,opticnerve trauma, human major blinging eye diseases,are based on the retinalganglion cells (retinal ganglion cells,RGCs) death and the fiber progressive death.Due to the non-renewablity after the injury,these diseases have no effectivetreatment.Ischemia and hypoxia is a common pathological features of retinaldiseases.Retinal Müller cells is the major glial cell in the retina,the RGCs rely onthe intracellular high affinity L-glutamate/L-aspartate transporter GLASTmaintain extracellular glutamate concentration,protect RGCs from glutamatecytotoxicity.Unfolded protein response(UPR) is the cells area in externalstimulity,the endoplasmic reticulum(ER) restores a large number of unfoldedprotein,triggered a series of reaction.Objective：1. Retinal Müller cells cultured in primary;2. To investigate the existence of unfolded protein response (UPR) in retinalMüller cells by hypoxia;3. To investigate therelationship between UPR related factors (XBP1andCHOP)andGLAST. Methods：1. Retinal Müller cells of rat at postnatal day3~7were cultured bytrypsin-dispersed cell culture;2. Cultured retinal Müller cells were maintained withCocl2/DTT for3h、6h、12h、24h、48h、72h respectively in vitro.Control：cultured in Fetal bovineserum medium;3. The expression of XBP1、CHOP and GLAST were quantified by Real TimePCR and Western-blot.Results：Under hypoxia condition，XBP1、CHOP and GLAST co-expressed inretinal Muller cells. The expression levels of the three factors were all ascendedand reached the summit after24h by hypoxia while dropped sharply after72h，which was consistant with the result of DTT.Conclusion：The cultured retinal Müller cells were positive in GS and GLASTantibody staining.Expression of GLAST and Unfolded Protein ReactionRelated Factors XBP1、CHOP were all acended in retinal Müller cell by hypoxiaand DTT.