The Individual And Combined Effects Of Vitamin C And Vitamin E On Rat Embryonic Cerebral Neurons Following Hypoxic Damage In Vitro

Objective To study the individual and combined effects of vitamin C(VC) and vitamin E(VE) on rat embryonic cerebral neurons following hypoxic damage in vitro.Methods The method of serum-free medium culture was applied in this study. The rat embryonic cerebral neurons were cultured with serum-free medium in which VC/VE was added. The neurons were divided into eleven groups. They were Normal control, Hypoxia, Hypoxia + VC-L,M,H(5,10, 25umol/L ), Hypoxia + VE-L,M,H(10, 25, 50umol/L), Hypoxia + combination of VCE-L,M,H(VC2.5umol/L+VE5umol/L,VC5umol/L+VE12.5umol/L, VC12.5umol/L+VE25umol/L) groups, respectively. The cells were incubated in 95% N2 for 30 minutes after they were incubated with VC or VE for three days. The growth, development and morphology of neurons were observed by using the phase contrast microscope. At the end of the experiment, cultured cerebral neurons were collected to detect bio-chemical indexes and make a pathological observation by using light and electronic microscope.Results (1)Compared with the Hypoxia group, the level of MTT, the activity of SOD, GSH-Px and NSE, contents of protein in other groups were higher significantly(P<0.01); and the contents of MDA, the releasing percent of LDH and the concentration of Ca2+ in other groups were lower significantly (P<0.01). The activity of ACHE in the Hypoxia group was lower significantly than that in other groups except the VC-L, M groups (P<0.01). (2) All the indexes above mentioned in the VE groups were better than that in the VC groups. (3) All the indexes except the activity of ACHE in the VCE-L group were significantly different with the VC-L group (P<0.01), but there was no significant difference between the VCE-L group and the VE-L group (P>0.05). Besides, there was significant difference about all the indexes between the VCE-M group and the VC-M group (P<0.01), alsobetween the VCE-H group and the VC-H group (P<0.01). The level of MTT, the activity of SOD, GSH-PX and NSE in VCE-M group was higher significantly than that in the VE-M group (P<0.01). Compared with the VE-H group, the level of MTT, the activity of SODs GSH-PX and NSE in VCE-H group was higher significantly(P<0.01), and the contents of MDA was lower significantly(P<0.01). (4) The results of light microscope showed that the neurons were distortion and the neurites were thin and short in the Hypoxia group. The networks between cell masses in Hypoxia group were sparse and scattered. The number of cells was more and the volume of cell masses was bigger in the VC-L group than that in the Hypoxia group. With the increase of the dose of VC, the cells' form obviously took a favorable turn. The cells in the VE groups restored being better. The neurons in the VE-L group took the form of fusiform or awl, and were basically similar with the Normal control group. The neurons in the VE-IvL H groups grew vigorously, the networks between the neurites in which were extremely dense. The results of electronic microscope showed that the neurons showed the broken appearance and the tiny floss of the cells was desert in the Hypoxia group, the cells in which lost the normal cell's life state. There was significant better recovery in the VC-H and VE-H groups. The three-dimensional effect of the cells is strong and the tiny floss of the cells was abundance in the VC-H and VE-H groups. The cell morphology of the VE groups was better than that in the VC groups.Conclusion The results showed that VC and VE could promote the recovery of cerebral neurons of rat embryonic following anoxic damage in vitro. Its role mechanism may be produced through scavenging free radicals directly, elevating the antioxidative activity in cells, and inhibiting the peroxidative reaction of cell membranes. The nueroprotective effect of VE is superior to VC. The combination of VC and VE has synergistic effect, the effect of which is better than that of using only one of them.