We aimed to investigate the effects of job stress on the biochemical variables of schoolteachers before and after stress reduction strategies. In 2019, 200 schoolteachers (98 males, 102 females) in the private and fully aided higher secondary schools of urban and rural areas in Vellore, India were enrolled in the study based on the principle of consecutive sampling. Questionnaires were used to investigate demographic characteristics and occupational stress-related factors. Fasting blood glucose was detected by venous plasma and a biochemical analyser with the cut-off thresholds method. Salivary cortisol was determined by the specific enzyme-linked immunosorbent assay kit method. Total cholesterol in serum was determined by the CHOD-PAP method. Serumalbumin was detected by the bromocresol green dye-binding method on a Roche Modular DP analyser (Roche Diagnostics). Blood pressure was measured by aneroid sphygmomanometers. Levels of fasting blood glucose, salivary cortisol, albumin and cholesterol were lower in the high-tension group than those in the low-tension group. Those of the high-coping strategy group were higher than those of the low-coping strategy group. Systolic blood pressure and diastolic blood pressure significantly reduced among the intervention groups after administering the relaxation response programme for three and six weeks, respectively, to urban and rural school teachers. Occupational stress can lead to increased salivary cortisol, cholesterol, albumin, blood glucose and blood pressure levels. The relaxation response programme resulted in quantifiable improvements in physiological markers of stress in rural and urban schoolteachers.