The Protective Effects Of Bone Marrow Mesenchymal Stem Cells On Hydrogen Peroxide Induced Retinal Ganglion Cells Injury

Objective To investigate retinal ganglion cells(RGC-5) apoptosis after hydrogen peroxide induced and protective effects of bone marrow mesenchymal stem cells(BMSCs) on hydrogen peroxide induced damage in RGC-5 cells. Method BMSCs were collected by the means of whole bone marrow adherent from SD rats, and test the surface markers through flow cytometric analysis. Immunocytochemistry was conducted to identify RGC-5 cells specific marker Thy1.1 expression. Exposure of RGC-5 cells to the indicated concentration of hydrogen peroxide(H2O2) for 24 h, analyzed by CCK-8 assay to choose an appropriate concentration to build H2O2 damage model. Morphological changes in RGC-5 cells were observed under microscope. The apoptosis of RGC-5 cells was detected by Hoechst fluorescence staining. BMSCs co-cultured with RGC-5 in a transwell culture system for 24 h and 48 h. The apoptosis rate of RGC-5 cells was examined by flow cytometry. The supernatant of RGC-5 cells was detected for TNF-?, IL-1? and BDNF concentration using ELISA. The activity of MDA and SOD were assayed by biochemistry technology.Result 1.BMSCs were successfully obtained by the means of whole bone marrow adherent. Flow cytometry showed that the 3th passage of BMSCs were positive against CD90 and CD44,while negative against CD34 and CD45. 2.After H2O2 exposure for 24 h and 48 h, the morphological varieties were surveyed as cytoplasm shrinking and paramorphia together with nuclear gathering. Reduced attachment and cell fragment were also identified. Compared with normal group, the count of apoptotic cells was added by Hoechst fluorescence staining. 3.After 200umol/l H2O2 induced for 24 h, BMSCs were co-cultured with RGC-5 cells for 24 h and 48 h. We found that apoptosis rate of cells was remarkable decreased. In contrast to H2O2 induced group, co-cultured group showed that inflammatory factors TNF-? and IL-1? reduced in supernatant of RGC-5 cells. Intra-cellular oxidant factor MDA decreased and anti-oxidant factor SOD increased. In the mean time, the expression of neurotrophic factors BDNF augmented.Conclusion H2O2 exposure induced damage to RGC-5 cells. BMSCs co-cultured with RGC-5 cells were able to reduce the apoptosis rate. It could be related with enhacing anti-oxidant capacity, suppressing inflammation, secreting neurotrophic factors.