ABSTRACT: Objectives of this study were: (1) to examine gender differences in biomedical indicators, lifestyle behaviors, self-health check practices, receipt of professional non-communicable disease (NCD)-related lifestyle advice, and the use of health services among teachers in Afghanistan; and (2) to seek the patterns of these indicators among users and non-users of health services among both male and female teachers. This cross-sectional study was carried out among 600 schoolteachers in Kabul city in February 2017. Gender differences in percentage distributions of abnormal biomedical indicators, lifestyle behaviors, self-health check practices, and receipt of professional lifestyle advice were examined. These patterns were further analyzed according to the use of health services in the previous 12 months by both genders. The results showed that male teachers had a higher prevalence of hypertension, increased serum triglycerides, physically active lifestyle, and tobacco use than female teachers (28.2/20.4, p = 0.038; 47.0/37.9, p = 0.040; 54.3/40.9, p = 0.002; 15.8/0.7, p < 0.001, respectively); female teachers had a higher prevalence of increased serum LDL cholesterol, overweight/obesity, and frequent consumption of fruits/vegetables than male teachers (61.3/50.8, p = 0.018; 64.7/43.5, p < 0.001; 71.4/53.8, p < 0.001, respectively). Female teachers were more likely to receive professional lifestyle advice related to NCDs than male teachers. Although users of health services practiced self-health checks and received professional lifestyle advice more frequently than non-users, abnormal biomedical indicators were similarly shown among users and non-users of health services in both genders. In conclusion, high prevalence of abnormal biomedical indicators was indicated in both male and female teachers, although the specific abnormal biomedical indicators differed by gender. Users and non-users of health services presented a similar prevalence of these abnormal indicators. Understanding the differences in patterns of NCD risk factors is essential when developing gender-informed policies.