miR-344b-3p Targeted Tspo Gene is Involved in Bisphenol S-induced Testosterone Synthesis Abnormal in Mouse
Objective To explore the role and mechanism of bisphenol S(BPS)affecting testosterone synthesis in mice.Methods Mouse models of BPS toxicity at concentrations of 0,2,10,and 50 mg·kg-1 were construct-ed.HE staining was used to observe the morphology of their testicular tissues,CASA was applied to analyze the sperm viability,and ELISA was used to detect the changes in serum testosterone levels.RT-qPCR and Western blot were used to detect the expression levels of mRNAs and proteins of key genes for testosterone production.The upstream miRNAs of the target genes of testosterone synthesis inhibition by BPS were predicted and screened using the mammalian miRNA target gene database(miRDB),and the expression levels of the candi-date miRNAs were detected by RT-qPCR in testicular tissues and TM3 cells of BPS-exposed mice.The In-hibitor/Mimics of candidate miRNAs were further constructed to transfect the TM3 cell model,and on this ba-sis,BPS exposure treatment was performed to detect the expression of the predicted target genes and changes in protein levels to identify the upstream miRNAs.Results By constructing mouse models with different concen-trations of BPS exposure,it was found that BPS exposure for 35 days resulted in histopathological damage to the testis,decreased sperm viability and decreased serum testosterone levels in mice(P all＜0.05).The combined RT-qPCR and Western blot results suggested that Tspo and Cyp11a1 were the target molecules of BPS-in-duced decrease in testosterone production in mice(P＜0.01).The upstream miRNAs of testosterone synthesis target genes were screened by miRDB database as miR-344b-3p and miR-124-5p,respectively.miR-344b-3p and miR-124-5p expression levels were found to be significantly up-regulated in BPS-treated groups compared with the control group in both in vivo and ex vivo models(P all＜0.01).Inhibitors and mimics of transfected miRNAs revealed that miR-344b-3p could regulate Tspo gene expression(P＜0.05).Further treatment with BPS found that transfection of mimics could suppress the expression level of Tspo gene and protein even more(P＜0.05).Conclusion BPS is involved in testosterone synthesis abnormal in mice by inhibiting the expres-sion of its downstream target gene Tspo via miR-344b-3p.
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