Objective To study the effect of cigarette smoke extract(CSE) on the diaphragm cells of rats in 8-hydroxydeoxyguanosine(8-OHdG) content and the relationship between 8-OHdG content and level of reactive oxygen species(ROS) and 8-oxoguanine DNA glycosylase-1(OGG1). Methods The diaphragm cells of rats were cultured and stimulated by CSE for different time periods and at different concentrations. 8-OHdG was detected by high performance liquid chromatography with electrochemical detection(HPLC-ECD). The level of ROS was determined by fluorescent probe of 2’, 7’-dichlorodihydrofluorescein diacetate (DCFH-DA). The expression of OGG1 mRNA was tested by real-time PCR. The expression of OGG1 protein was tested by Western blot. Results There was no significant difference in the 8-OHdG, ROS, OGG1 mRNA and protein levels after CSE treatment for different time periods. However, the 8-OHdG content of the diaphragm cells induced by 10.00% CSE and 20.00% CSE was significantly increased compared with the diaphragm cells induced by 5.00% CSE and control groups(P＜0.05). There was no significant difference in 8-OHdG content between 5.00% CSE and control groups(P＞0.05). Compared with control, the intracellular ROS level was dramatically increased in CSE-treated groups(P＜0.05), and the 8-OHdG content was increased according to the level of ROS(r＝0.826, P＝0.000). Compared with control, the expression of OGG1 mRNA and protein were dramatically increased in CSE-treated groups(P＜0.05), however, the 8-OHdG content has but not with the expression of OGG1 mRNA and protein(r＝0.373, P＝0.254; r＝0.329, P＝0.296). Conclusions CSE could increase the level of 8-OHdG in the diaphragm cells of rats. The 8-OHdG content has correlation with ROS level, but not with the expression of OGG1.