Bisphenol A Inhibits Testosterone Synthesis in TM3 Cells by Upregulating Apoa1 Gene Expression
The aim of the study was to investigate whether apolipoprotein A1(apolipoprotein A1,Apoa1)mediates the reduction of testosterone synthesis in mouse Leydig cells line(TM3)induced by bisphenol A(BPA)exposure from the perspective of lipid metabolism.TM3 cells were randomly divided into 7 groups with different concentrations(0,5,10,20,40,60,and 80 μmol·L-1),and 0 μmol·L-1 BPA was the control group(CON);After 24 h of treatment with the different concentrations,cell viability was detected by CCK-8 method to determine the optimal dose of BPA.Testosterone(testosterone,T)content in TM3 cell supernatant was detec-ted by ELISA;mRNA expression levels of lipid metabolism-related genes Apoa1,Apoa2(apoli-poprotein A2)and Apoc3(apolipoprotein C3)genes were measured in TM3 cells by RT-qPCR.APOA1 protein expression level was detected by Western blot and immunofluorescence method.Intracellular lipid droplet accumulation was observed by oil red O staining.The results showed that 20 μmol·L-1 BPA treatment for 24 h had no significant effect on the viability of TM3 cells;However,an extremely significant inhibition of TM3 cell viability was observed in treatment with 40 μmol·L-1 BPA for 24 h(P＜0.01);In addition,after 20 μmol·L-1 BPA treatment on TM3 cells for 24 h,the testosterone content in the culture supernatant was extremely significantly lower than that in the CON group(P＜0.01),the mRNA expression level and protein expres-sion of Apoa1 gene were extremely significantly elevated(P＜0.001),but there was no signifi-cant change in the mRNA expression level of Apoa2 or Apoc3;The accumulation of lipid drop-lets in TM3 cells was extremely significantly reduced by 20 μmol·L-1 BPA treatment for 24 h compared with the CON group(P＜0.000 1).In conclusion,BPA can reduce the lipid droplets accumulation in TM3 cells by up-regulating Apoa1 expression levels,enhancing reverse choles-terol transport(RCT),leading to a decrease in testosterone synthesis in TM3 cells.
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