The downregulation of PEPCK mediated cuproptosis is involved in triclosan induced reproductive damage in mouse testes via triggering ROS overproduction.

Triclosan (TCS) is a widely used broad-spectrum antimicrobial agent that has been demonstrated to induce damage to the male reproductive system. However, the specific mechanisms of testicular injury induced by TCS are not fully understood. In the present study, mice were treated with 0, 20 mg/kg and 40 mg/kg per day TCS from PND 56 to PND 84. Decreased sperm counts, increased sperm deformity rates, downregulation of Phosphoenolpyruvate carboxykinase (PEPCK) and cuproptosis activation were observed. In vitro, TM4 cells were exposed to 0, 5 nM and 10 nM TCS for 24 h, which induced oxidative stress damage, inhibition of PEPCK and cuproptosis activation. Furthermore, inhibition of PEPCK and cuproptosis activation was rescued by reactive oxygen species (ROS) scavengers N-Acetylcysteine (NAC). Besides, cuproptosis activation induced by TCS were rescued after overexpression of PEPCK. Drug prediction with Connectivity Map (cMap) identified flubendazole and cucurbitacin-i as potential therapeutic drug. In conclusion, this study demonstrated that TCS triggered cuproptosis activation in TM4 cells mediated by ROS storm induced PEPCK inhibition. This research also shed new insight into precise treatment of TCS-related testicular injury.