农药百克敏对小鼠精母细胞GC-2铁死亡的诱导作用OA

Objective:To discuss the effect of pyraclostrobin on ferroptosis in the spermatocytes GC-2 of the mice,and to clarify whether pyraclostrobin can cause male reproductive toxicity.Methods:The mouse spermatocytes GC-2 were divided into control group,low dose of pyraclostrobin group(0.8 µmol·L-1 pyraclostrobin),medium dose of pyraclostrobin group(1.6 µmol·L-1 pyraclostrobin)and high dose of pyraclostrobin group(2.4 µmol·L-1 pyraclostrobin).After being treated with pyraclostrobin for 24 h,methylthiazolyldiphenyl-tetrazolium bromide(MTT)method was used to detect the proliferation activities of the GC-2 cells after treated with different doses of pyraclostrobin;mitochondrial membrane potential detection kit JC-1 was used to detect the mitochondrial membrane potential in the GC-2 cells in various groups;superoxide dismutase(SOD)activity detection kit was used to detect the activities of SOD in the GC-2 cells in various groups;malondialdehyde(MDA)assay kit was used to determine the levels of MDA in the GC-2 cells in various groups;reduced glutathione(GSH)and oxidized glutathione(GSSG)detection kits were used to detect the levels of GSH and GSSG in the GC-2 cells,and the ratios of GSH/GSSG in various groups were calculated;Western blotting method was used to detect the expression levels of heme oxygenase-1(HO-1),ferritin heavy chain 1(FTH1)and glutathione peroxidase 4(GPX4)in the GC-2 cells in various groups.Results:Compared with control group,the proliferation activities of the GC-2 cells in various pyaclostrobin dose≥1.0 μg·L-1 groups were decreased(P＜0.01).The low,medium,and high doses of pyraclostrobin were 0.8,1.6,and 2.4 µmol·L-1.The results of JC-1 method showed that compared with control group,the mitochondrial membrane potential of the GC-2 cells in medium,and high doses of pyraclostrobin groups were decreased.Compared with control group,the SOD activity in the GC-2 the cells in low,medium,and high doses of pyraclostrobin groups were decreased(P＜0.01),and the MDA levels in the GC-2 cells were increased(P＜0.01).Compared with control group,the GSH levels in the GC-2 cells in low,medium,and high doses of pyraclostrobin groups were significantly decreased(P＜0.01),the GSSG levels were increased(P＜0.01),and the GSH/GSSG ratios were decreased(P＜0.01).Compared with control group,the expression levels of HO-1 protein in the GC-2 cells in low,medium,and high doses of pyraclostrobin groups were significantly increased(P＜0.05 or P＜0.01),while the expression levels of FTH1 and GPX4 proteins were significantly decreased(P＜0.05 or P＜0.01).Conclusion:Pyraclostrobin can induce the ferroptosis of the spermatocytes GC-2.