中国药理学与毒理学杂志2024，Vol.38Issue(6)：426-435,10.DOI:10.3867/j.issn.1000-3002.2024.06.004
三氯异氰尿酸通过诱导氧化应激和铁死亡抑制精原细胞增殖
Trichloroisocyanuric acid inhibits spermatogonia proliferation by inducing oxidative stress and ferroptosis
摘要
Abstract
OBJECTIVE To explore the effects of trichloroisocyanuric acid(TCCA)on the prolifera-tion of spermatogonia by inducing oxidative stress and ferroptosis.METHODS GC-1 cells were cultured in DMEM-F12 medium,and cell proliferation was plotted according to the growth curve.GC-1 cells were treated with TCCA at concentrations of 0(cell control),97,194,and 387 μmol·L-1 for 24 h.Cell viability was detected using the CCK-8 method,apoptosis cells were stained with Hoechst 33342,cell cycle was examined by PI staining method,RT-qPCR was performed to measure the mRNA expres-sion levels of apoptosis-related genes Bax,Fas,oxidative stress-related genes superoxide dismutase 2(SOD2),glutathione peroxidase 4(GPX4),nuclear factor erythroid 2-related factor 2(Nrf2),solute carrier family 7 member 11(SLC7A11),dihydroorotate dehydrogenase(DHODH),DNA methyltransferase 3A(DNMT3A),and DNA methyltransferase 3L(DNMT3L).The Griess method was used to determine the nitric oxide(NO)content,colorimetric method for the malondialdehyde(MDA)level,DCFH-DA fluores-cence probe method for the reactive oxygen species(ROS)level,DNTB colorimetric method for the reduced glutathione(GSH)content,and WST-8 method for the reduced coenzymeⅡ(NADPH)content.RESULTS Compared with the cell control group,the cell survival rates in the TCCA 194 and 387 μmol·L-1 groups decreased significantly(P<0.01),accompanied by nuclear condensation and fragmentation,a significant increase in apoptosis rate(P<0.01),and cell arrest in the G2/M phase(P<0.05).Additionally,in the TCCA 387 μmol·L-1 group,the levels of NO,MDA and ROS increased(P<0.01),while the levels of GSH and NADPH decreased(P<0.01).Moreover,the mRNA expressions of SOD2,GPX4,Nrf2,SLC7A11,and DHODH decreased(P<0.05,P<0.01),while the expressions of Bax,Fas,DNMT3L,and DNMT3A increased(P<0.05,P<0.01).CONCLUSION TCCA exposure reduces the viability of GC-1 cells,inhibits cell proliferation,induces apoptosis of GC-1 cells.The mechanism may be related to the ability of TCCA to enhance oxidate stress,induce ferroptosis,and interfere with the methylation of GC-1 cells.关键词
三氯异氰尿酸/精原细胞/氧化应激/DNA甲基化/铁死亡Key words
trichloroisocyanuric acid/spermatogonia/oxidative stress/DNA methylation/ferroptosis分类
医药卫生引用本文复制引用
蒋莉,韩雪,吴德生,黄海燕,刘建军..三氯异氰尿酸通过诱导氧化应激和铁死亡抑制精原细胞增殖[J].中国药理学与毒理学杂志,2024,38(6):426-435,10.基金项目
深圳市医学重点学科建设经费资助(SZXK069) Shenzhen Key Medical Discipline construction Fund(SZXK069) （SZXK069）
