The Effects Of Vitamin A Metabolic Abnormality On The Development Of Leydig Cells In Rats

Objective:To explore the effects of vitamin A metabolic abnormality on the development and differentiation of Leydig cells in rats,we carried out two experiments from two aspects(vitamin A deficiency and vitamin A metabolic enzyme Alcohol dehydrogenase 1 expression abnormality).Method:1)Vitamin A deficiency food was used to feed the female SD rats during lactation,who breastfeed rat pups.Then the offspring rats continued to use Vitamin A deficiency food after ablactation.At postnatal day 14,day 35,day 90,male rats were sacrificed,then testis and serum were collected.The concentration of serum testosterone was detected by radiation immunology technology.The testosterone synthesis enzymes related gene(Hsd17b3)in mRNA level was detected by real-time fluorescence quantitative PCR.The protein level of Vitamin A catabolic enzymes(ADH1?ALDH1A1)and testosterone synthesis enzymes(NR5A1?STAR?CYP11A1?HSD3B)were detected by western blotting.2)Adult male SD rats were injected intraperitoneally with EDS to establish leydig cell regeneration model,then treatment with ADH1 inhibitor 4-methylpyrazole(4-MP)each day.At day 14,21,and 28,these rats were sacrificed.The concentration of serum testosterone was detected by radiation immunology technology.The protein level of Vitamin A catabolic enzymes(ADH1?ALDH1A1)and testosterone synthesis enzymes(NR5A1?STAR?CYP11A1?HSD3B)were detected by western blotting.Result:1)Vitamin A deficiency affected the increase of body and testis weight of SD rats,and inhibited the expression of testosterone synthesis related genes and proteins of leydig cells,which leaded to a significant lower serum testosterone.2)In leydig cell regeneration model,ADH1 inhibitor 4-methylpyrazole(4-MP)could specifically inhibit the protein expression of ADH1 of the testis and did not change the protein expression of ALDH1A1.After treated with 4-methylpyrazole(4-MP)for 14 days,21 days,and 28 days,the results suggested that down-regulation the protein expression of ADH1 could significantly inhibit the protein expression of NR5A1 and testosterone synthesis related enzymes,leading to inhibition the regeneration of leydig cell and serum testosterone recovery.Conclusion:Vitamin A had an important effect on the development of leydig cell.The abnormal metabolism of vitamin A affected the development and function of leydig cell.