Inhibition of H_(2)O_(2)-induced apoptosis of GC2-spg cells by functionalized selenium nanoparticles with lentinan through ROSmediated ERK/p53 signaling pathways

AH_(2)O_(2)-induced oxidative stress injury cell model was established to investigate the antioxidant effect of nanoselenium on mouse spermatocyte lines and the regulation mechanism of the expression level and activity of seleniumcontaining antioxidant enzymes induced by oxidative stress.A safe and effective nano-drug system of functionalized selenium-containing nanoparticles(SeNPs)was developed with lentinan(LNT)(SeNPs@LNT).Mice spermatocyte line GC2-spg cells were treated with SeNPs@LNT(1,2,4,8,16,32μM)for 24-72 h to evaluate the cytotoxicity of selenium.GC2-spg cells were randomly divided into the following groups:control,hydrogen peroxide(H_(2)O_(2)),SeNPs@LNT,and H_(2)O_(2)+SeNPs@LNT groups.H_(2)O_(2)+SeNPs@LNT group was pretreated with SeNPs@LNT 4μM for 12 h,followed by H_(2)O_(2)600μM for 8 h.The cell viability decreased in the H_(2)O_(2) group and increased significantly in the SeNPs@LNT group.Compared with the H_(2)O_(2) group,the SeNPs@LNT+H_(2)O_(2) group exhibited obvious red fluorescence,indicating a higher level of mitochondrial membrane potential.The content of intracellular reactive oxygen species(ROS)in the SeNPs@LNT group reduced significantly,and the intensity of green fluorescence in the SeNPs@LNT+H_(2)O_(2) group decreased significantly compared with the H_(2)O_(2) group,indicating the inhibitory effect of SeNPs@LNT on the generation of ROS-induced oxidative stress.The activity of GPx and SOD increased significantly in the SeNPs@LNT group.The expression of p53 decreased significantly under the intervention of nano-selenium,and GPx1 expression increased.In the oxidative stress group,the expressions of DNA damage-related proteins and apoptosis-related proteins were higher than those in other groups.Thus,SeNPs@LNT can promote GC2-spg cell proliferation,improve GPx and SOD activities,remove intracellular ROS,and reduce mitochondrial damage and functional abnormalities caused by oxidative stress by regulating the ERK and p53 protein levels.SeNPs@LNT has good biological activity and antioxidant effect,which can be used to protect the male reproductive system from oxidative stress.