Study The Mechanism Of Protective Effect Of Astragalus Membranaceus Injection On Oxidative Damage To Müller Cell

Object:To study the protective effect of Astragalus membranaceus injection on human retinal Müller cells that H₂O₂ hadinduced to be injured and the mechanism of anti- apoptosis.Methods:By culturing human retinal Müller cell in conventional methods, it is to establish the cell injured model of H2O2 on Müller cells. The experiments were divided into blank control group, model group, AMI groups from low to high, 3 dose groups（10 mg/L, 30 mg/m L, 90mg/m L）, The methods of MTT was to observe the effects of different concentrations of AMI on each Müller cells viability; cell apoptosis was detected by flow cytometry; Western blot was to detect Caspase-3 and NF- κB proteins of in each group of Müller cells; RT-PCR assay was to detect Caspase-3, intracellular NF-κB m RNA expression in each group of Müller cells.Results:1.MTT results showed that: compared with the control group（0.94±0.02） comparison, model group（0.43±0.01） cell survival rates were decreased significantly（P<0.01）;compared with the model group, AMI groups（low, medium, high concentration group were 0.47±0.02, 0.56±0.04, 0.71±0.03） cell survival rates were increased significantly（P<0.05）.2.The results of flow cytometry apoptosis display: compared to the blank control group（4.06%）,the cell apoptosis rate of model group（41.81%） increased significantly（P<0.01）, the experimental drug group（low, medium, high concentration group were 32.95%, 24.52%, 18.49%） compared with the control group, cell apoptosis rate increased significantly（P<0.01）, and the AMI groups in experimental groups were compared with the model group to show the apoptosis rate decreased to different degree（P<0.05）.3.Western blot for detecting protein expression showed that: compared with the blank control group, the expression of model groups of Caspase-3 and NF- k B protein were significantly increased（P<0.01）, Compared with the model group, the expression of the AMI groups Caspase-3, NF- k B proteins significantly decreased（P<0.05）. 4.RT-PCR detect the expression of m RNA results show that: compared with the blank control group, the expression of model groups of Caspase-3 and NF- k B m RNA were significantly increased（P<0.01）, Compared with the model group, Caspase-3, NF- k B m RNA expression of AMI groups decreased obviously（P< 0.05）.Conclusion:Experimental results show that AMI can significantly inhibited oxidative damage caused by H2O2 on the Müller cells, improve the survival rate of the Müller cells, reduce the rate of cell apoptosis and decrease the expression of Caspase-3 and NF-k B protein and m RNA. The mechanism may be the effective components of AMI effect on the apoptotic response process of the key regulatory factors and cell’snuclear transcription factor NF- k B induced by upstream signaling pathways, and so the anti-oxidative damage and anti-apoptosis were played.