Minocycline inhibits rat retinal neural cell apoptosis induced by glutamate

AIM:To investigate the anti-apoptotic effect of minocycline on the rat retinal neural cells.METHODS:Primary cultures of retinal neural cells were prepared from neonatal SD rats.The expressions of NSE(neuron),GFAP(glial cells)and lectin(microglia)were detected by immunocytochemistry.Cultured cells were divided into normal control group(group A),minocycline control group(minocyline 20 μmol/L,group B),glutamate control group(glutamate 1 mmol/L,group C)and minocycline-treating group(minocycline 20 μmol/L+glutamate 1 mmol/L,group D).After 1 h intervention,Annexin V/PI flow cytometry was performed to evaluate the apoptotic cells with Annexin V/PI and mitochondria membrane potential(MMP)with Rh123.After 20 h intervention,MTT was used to test the cell viability,and culture supernatant was collected to test NOS activity.RESULTS:Annexin V/PI testing revealed that the cell apoptotic rate was 5.1% in group A,4.3% in group B,15.2% in group C and 8.3% in group D.MTT showed that mean absorbance was 0.093 in group A,0.099 in group B,0.038 in group C and 0.088 in group D.No significant difference between group A and group D was observed.Significant difference between group C and other three groups was found.MMP in group C was reduced compared with group A,which was reversed by minocycline treatment in group D.If NOS activity in group A was 1,that in group B was 0.987±0.219,in group C was 1.513±0.472 and in group D was 1.176±0.259,a significant difference between group C and other three groups was observed.CONCLUSION:Minocycline significantly decreases retinal apoptosis in neural cells treated with glutamate.The mechanism is related to suppressing NOS activity,and stabilizing the MMP in the cells.