Glutathione persulfide prevents high-fat diet induced down-regulation of testosterone biosynthesis
Objective To investigate effects and underlying mechanisms of glutathione persulfate(GSSH)on the level of testosterone in male obese mice.Methods Totally 45 mice were divided into 3 groups on average.Low-fat diet(LFD)+normal saline(NS)group:15 mice were fed with LFD for 10 weeks,followed by LFD together with daily intraperitoneal injection of saline for 45 d;high-fat diet(HFD)+NS group:15 mice were fed with high-fat diet for 10 weeks,followed by HFD and daily intraperitoneal injection of NS for 45 d;HFD+GSSH group:15 mice were fed with HFD for 10 weeks,followed by a HFD for 45 d and daily intraperitoneal injection of GSSH(200 mg/kg).After the treatment,all mice were killed with their necks-severed,testis and serum were taken out from the mice.Serum levels of testosterone and malondialdehyde(MDA),the mRNA levels of key enzymes for testosterone synthesis(StAR,3β-HSD,Cyp11a1 and Cyp17a1)were measured by RT-PCR.The testicular protein levels of StAR,3β-HSD,NR5A1 and EHD3 were measured by Western blotting assay.Protein levels of NR5A1,SOD and Nrf2 were measured in mouse Leydig TM-3 cells that were treated with 50 μmol/L and 100 μmol/L GSSH,respectively,following with treatment with 100 μmol/L H2O2.Results 1)After treatment,the body weight of mice in HFD+GSSH group did not change significantly,while the body weight of mice in HFD+NS group raised by 24.53%(from 32.46 g to 40.43 g)during the 45-day-intraperitoneal injection(P=0.002).2)Serum level of testosterone in HFD+NS group[(12.9±1.7)μg/L]was significantly lower than that in LFD+NS group[(18.3±1.2)μg/L,P=0.020].However,serum level of testosterone in HFD+GSSH group was(25.42±2.1)μg/L,which was significantly higher than that in HFD+NS group(P=0.030).The RT-PCR test results showed that compared with LFD+NS group,the expression levels of all key genes involved in testosterone synthesis(StAR,3β-HSD,Cyp11a1,Cyp17a1)showed a significant decrease in HFD+NS group(P=0.003,P=0.007,P<0.001,P<0.001).The expression levels of these genes were restored in the mouse testes of HFD+GSSH group(P=0.002,P<0.001,P<0.001,P=0.006).3)Similarly,compared with LFD+NS group[(9.00±1.59)nmol/mL],the serum MDA level of HFD+NS group[(10.61±1.73)nmol/mL]raised significantly(P=0.016),while GSSH reversed the raised HFD+NS high level of serum MDA in HFD+GSSH group[(9.23±0.94)nmol/mL,P=0.048].4)Both levels of NR5A1,EHD3,StAR,and 3β-HSD were reduced in HFD+NS group(P=0.002,P=0.012,P=0.004,P=0.043),but their levels were significantly restored in HFD+GSSH group(P<0.001,P=0.017,P=0.004,P<0.001).5)The levels of NR5A1,Nrf2 and SOD were obviously down-regulated in TM3 cells treated with H2O2(P<0.001,P=0.002,P=0.004).Conclusion GSSH can raise serum level of testosterone in HFD-fed mice by up-regulating expression of genes which are important for testicular testosterone biosynthesis.
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