〔Abstract〕 Objective To investigate the mechanism of sertoli/spermatogenic injury induced by cigarette smoke extract(CSE) based on ROS/NLRP3 signaling pathway.Methods The sertoli/spermatogenic of rat were cultured in vitro,and they were divided into control group,cigarette Smoke extract(CSE) group and CSE+NLRP3 inflammosome inhibitor MCC950 group.Cell viability was detected by CCK-8 assay;Lactate dehydrogenase(LDH) was used to detect cell membrane damage;reactive oxygen species(ROS) production was used to detect oxidative stress level;Hoechst/PI fluorescent staining was used to detect cell pyroptosis.Real-time quantitative PCR(RTqPCR) was used to determine the mRNA level of NLRP3,ASC,Caspase-1,GSDMD,IL-1β and IL-18.Western blot was used to determine the protein level of NLRP3,ASC,Caspase-1,GSDMD,IL-1β and IL-18.Results CCK-8 assay showed that cell viability decreased in CSE group(P<0.001),LDH leakage rate(P<0.001) and ROS(P<0.001) production increased.Compared to the CSE group,cell viability increased(P<0.001),LDH leakage rate(P<0.01) and ROS(P<0.001) production decreased in CSE+MCC950 group.The number of PI staining cells in CSE group was more than that in control group and CSE+MCC950 group,but staining cells in CSE+MCC950 group was less than that in CSE group.RT-qPCR and Western blot results showed that the expression levels of NLRP3,ASC,Caspase-1,GSDMD,IL-1β and IL-18 increased in CSE group but decreased in CSE+MCC950 group.Conclusion The oxidative stress and inflammation of sertoli/spermatogenic induced by cigarette smoke may be related to the activation of ROS/NLRP3 signaling pathway.