Oral Folic Acid And Betaine To Improve The Quality Of Sperm In Mice And Mechanism Of Action

With the pollution of the environment in recent years,the adjustment of the dietary structure and the changes in lifestyles and other factors,the sperm quality of men has been declining.Although assisted reproductive technology is now becoming more mature,there is no standard treatment plan for male infertility to stay at the level of empirical treatment.Folic acid and betaine are closely linked in body metabolism.Betaine can be used as a methyl donor and folic acid as a carrier for methyl.Both of them participate in a carbon metabolism.Previous studies have shown that folic acid can effectively scavenge hydroxyl radicals and improve sperm quality by reducing homocysteine toxicity,reducing DNA methylation levels,and reducing sperm oxidation.Betaine can improve the antioxidant capacity of testicular tissue by eliminating superoxide,and can produce S-adenosylmethionine through the betaine cycle to improve sperm motility and sperm function.Objective:To study the effects of folic acid and betaine on sperm quality and related molecular mechanisms,and to pave the way for further research on the pharmacological effects of folic acid and betaine and related functional foods.Methods:Kunming mice were divided into blank group,folic acid group(1mg/Kg),betaine group(60 mg/Kg),folic acid+betaine group(folate 1 mg/Kg,betaine60 mg/Kg).Six weeks after intragastric administration,microscopic examination was performed to examine the number,vigor,and malformation rate of spermatozoa.The test kits were used to detect the expression of SOD,MDA,and GSH-Px in the testis,and folic acid was evaluated.Effects of Betaine on Sperm Quality.Furthermore,the expression levels of Erk1/2 and Ras mRNA in mouse testis were detected using a real-time fluorescence quantitative PCR instrument at the nucleic acid level,and the expression of p-Erk1/2 and Ras protein in the testicular tissue was detected from the protein level using western-blot.Study its internal mechanisms.Results:The sperm counts(10~7 cells/ml)in the blank group,folic acid group,betaine group,and folic acid+betaine group were 1.352±0.06,1.35±0.05,1.8675±0.07,and 2.115±0.07,respectively.Compared with the control group,the amount of sperm in the betaine group was significantly increased(p<0.05),and the number of sperm in the folic acid+betaine group was significantly increased(p<0.001).The sperm motility(%)in the blank group,folic acid group,betaine group,and folic acid+betaine group was 64.74±3.14,74.81±4.67,80.27±3.98,and 75.79±4.25,respectively,and the folic acid group and folic acid were compared with the blank control group.The sperm motility in the betaine group increased significantly(p<0.05),the sperm motility in the betaine group increased significantly(p<0.01),and the sperm abnormality rate in the folic acid group,the betaine group,and the folic acid+betaine group was compared with the blank group.There was no significant difference(p>0.05).Compared with the blank group,the levels of T-SOD and GSH-Px in the folic acid group,the betaine group,the folic acid+betaine group were significantly increased,and the MDA content was significantly decreased.Fluorescent quantitative PCR results showed that the mRNA levels of Erk and Ras in the folic acid group,the betaine group,and the folate+betaine group were significantly higher than those in the blank group.The significance analysis results were p<0.05,p<0.001,and p<0.001,Western blot results showed that folic acid,betaine and folic acid+betaine can significantly or very significantly increase the expression of p-Erk1/2 and Ras protein,the difference was significant in each group p<0.01,p<0.001 and p<0.001.Conclusion:The results of this study indicate that folic acid and betaine can increase the number of spermatozoa and improve sperm motility,and neither of them can alter the sperm deformity rate;folic acid and betaine can increase the content of SOD and GSH-Px in the testis,and decrease the content of MDA.Content;Folic acid and betaine can increase the nucleic acid and protein levels of Erk1/2,Ras in testicular tissue.Therefore,folic acid and betaine may improve sperm quality by increasing the antioxidant capacity of testicular tissue and activating the Erk signaling pathway.Preliminary studies shed light on its possible molecular mechanism.