Protective effect of verbascoside on brain glial cells with oxidative stress injury

Abstract

Objective To investigate the protective effect of verbascoside on brain glial cells (HEB) with oxidative stress injured. Methods HEB cells were cultured by dulbeeeo modified eagle medium (DMEM) culture liquid containing 10% fetal bovine serum and randomly divided in to model group, verbascoside low, medium and high dose group, positive drug group and normal control group. Except normal control group, the other group were induced to oxidative stress injury by bing exposed to H2O2 at dose 100 mmol/L. 20, 40, 80 μg/L verbascoside were separately added into low, medium and high doses of verbascoside groups. Afer co-culturing for 4 h, the morphological changes of HEB were observed by light microscope; the survival rate of HEB was examined using methyl thiazol tetrazolium (MTT). The activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in the HEB were detected with spectrophotometry. The levels of lactate dehydrogenase (LDH) in culture medium were determined by enzyme linked immunosorbent assay (ELISA). Statisticalanalysis of data using the statistical product and service solutions 17.0 software. Results Compared with the normal control group, HEB in model group shrank, more suspension cells were in the culture medium, Compared with the model group, HEB cells in each concentration group and positive drug group retracted less and the number of cells increased significantly. The cell survival rate of model group was (38±6)%. The cell survival rates of low, medium and high concentration of pistil glycoside were (45±2)%, (63±4)% and (83±7)%, respectively. The cell survival rate of positive drug group was (83±2)%. Compared with the model group, the cell viability of the three concentration groups and the positive drug group increased significantly (t=3.120, 3.920, 0.892, P<0.01). In the normal control group, SOD level was 1.83±0.64, MDA level was 6.25±0.47 and LDH concentration in cell culture medium was 1357.78±110.24. In model group, SOD level was 0.94±0.21, MDA level was 14.35±1.17 and LDH concentration in cell culture medium was 1 994.32±88.25. Compared with the normal control group, SOD level of HEB cells in model group decreased (t=4.289, P<0.01), MDA level increased (t=3.204, P<0.01), LDH concentration in cell culture medium increased (t=3.056, P<0.01). SOD levels were 1.56±0.41, 1.86±0.74, 1.78±0.47, MDA concentrations were 8.06±0.43, 6.84±0.91, 6.63±0.25, LDH concentrations in cell culture medium were 1 548.36±109.47, 1 384.39±107.95, 1 297.81.36±114.32, respectively. Compared with the model group, the SOD level of middle dose of piloside increased (t=2.891, P<0.05), MDA concentration decreased (t=2.942, P<0.05), LDH concentration in cell culture medium decreased (t=2.877, P<0.05), SOD level increased (t=3.128, 3.762, P<0.01), MDA concentration decreased (t=3.320, 3.265, P<0.01) and LDH concentration in cell culture medium decreased (t=3.708, 3.962, P<0.01) in high dose group and positive drug group. Conclusion Mulberry glycoside can protect HEB from oxidative stress, and the protective mechanism may be related to improving the antioxidant capacity of HEB. Key words: Verbascoside; Oxidative stress; Brain glial cells