Tannic Acid Alleviate Oxidative Stress Induced by Sodium Fluoride in Mouse Oocytes
[Objective]The aim of this experiment was to study the protective effect of tannic acid(TA)on oxidative stress injury of mouse oocytes induced by sodium fluoride(NaF),and provide theoretical basis for developing antidotes for fluorosis.[Method]Mice oocytes were added with 0(control),0.25,0.50 and 0.75 mmol/L NaF in vitro maturation medium.The optimal concentration of NaF was determined by detecting the first polar body excretion(PBI)rate of oocytes.After treating mouse oocytes with the optimal concentration of NaF,the optimal concentration of TA was determined by adding different concentrations of TA(0,1,10 and 100 μg/L).On the basis of the above tests,control group,toxicity group(NaF)and antidote group(NaF+TA)were set up.The levels of reactive oxygen species(ROS),glutathione(GSH)and mitochondrial membrane potential(MMP)in mouse oocytes were detected by immunofluorescence staining.The fertilization rate,cleavage rate and blastocyst rate of mouse oocytes in each group were measured.The content of malondialdehyde(MDA)and the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were measured.[Result]Compared with control group,the excretion rate of the first polar body in 0.25-0.75 mmol/L NaF groups was extremely significantly decreased(P＜0.01).Then 0.25 mmol/L NaF was used as the effective toxic dose for testing.After NaF treatment,the excretion rate of first polar body in 10 μg/L TA group was extremely significantly higher than that in 0 μg/L TA group(P＜0.01).The results of immunofluorescence detection showed that,compared with control group,the ROS levels of oocytes in NaF group were significantly increased(P＜0.05),while the levels of GSH and MMP and the activities of SOD and GSH-Px were significantly or extremely significantly decreased(P＜0.05 or P＜0.01).Compared with NaF group,the levels of GSH and MMP and the activities of SOD and GSH-Px of mouse oocytes in NaF+TA group were significantly or extremely significantly increased(P＜0.05 or P＜0.01),ROS level and MDA content were significantly or extremely decreased(P＜0.05 or P＜0.01).The statistical results of early embryonic development showed that the fertilization rate,cleavage rate and blastocyst rate in NaF group were extremely significantly lower than those in control group(P＜0.01).The fertilization rate,cleavage rate and blastocyst rate of mice in NaF+TA group were extremely significantly higher than those in NaF group(P＜0.01),but had no significant difference compared with control group(P＞0.05).[Conclusion]NaF could induce oxidative stress in mouse oocytes by increasing ROS and MDA levels in oocytes,and reduce MMP levels,which affect early embryonic development.The addition of 10 μg/L TA could relieve the oxidative stress induced by NaF in mice and improve the maturation quality of mouse oocytes,thereby promoting in vitro fertilization and embryo development potential.
万方数据
维普
