Kumboyono Kumboyono, Indah Nur Chomsy, Cholid Tri Tjahjono, Yuliana Ratna Kumala, Titin Andri Wihastuti
Purpose: To examine simultaneous alterations in nitric oxide (NO), malondialdehyde (MDA), nicotinic acetylcholine receptors (nAChRs), and cotinine levels in smokers compared with non-smokers, and to clarify early mechanistic pathways linking tobacco exposure with vascular-related and oxidative stress. Patients and Methods: A cross-sectional study was conducted among 200 adults (100 smokers, 100 non-smokers) meeting predefined eligibility criteria and free from major cardiometabolic disease. Venous blood samples were analyzed for NO, MDA, nAChRs, and cotinine using standardized ELISA methods. Anthropometric and hemodynamic measurements were obtained using validated procedures. Group differences were assessed using independent t-tests and chi-square tests. Correlations were evaluated using Pearson or Spearman coefficients. Multivariable linear regression models adjusted for age, BMI, systolic and diastolic blood pressure to determine the independent association of smoking with biomarker profiles. Results: Smokers demonstrated significantly lower NO levels (70.22 ± 9.54 vs 198.27 ± 13.85 µmol/L; p < 0.001) and markedly higher concentrations of MDA (39.48 ± 4.54 vs 19.72 ± 5.32 nmol/mL; p < 0.001), nAChRs (20.19 ± 3.25 vs 12.05 ± 2.13 ng/mL; p < 0.001), and cotinine (61.07 ± 8.84 vs 4.99 ± 1.95 pg/mL; p < 0.001). After adjustment, smoking remained a strong independent predictor of reduced NO (β = −128.29; p < 0.001) and elevated MDA (β = 19.89; p < 0.001), nAChRs (β = 8.03; p < 0.001), and cotinine (β = 55.50; p < 0.001). These findings indicate persistent oxidative, nitrosative, and receptor-mediated biochemical disturbances associated with smoking, irrespective of baseline physiological differences. Conclusion: Smokers exhibit a distinct biochemical pattern characterized by reduced NO bioavailability, heightened oxidative stress, increased nAChR activation, and elevated cotinine levels. These alterations reflect early vascular-related biochemical disturbances associated with smoking and support the utility of these biomarkers for early cardiovascular risk detection. © 2026 Kumboyono et al.
Department of Community Health Nursing, Faculty of Health Sciences, Universitas Brawijaya, Malang, 65151, Indonesia; Biomedical Science Masters Study Program, Faculty of Medicine, Universitas Udayana, Bali, Indonesia; Department of Cardiology and Vascular Medicine, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia; Department of Conservative Dentistry, Faculty of Dentistry, Universitas Brawijaya, Malang, Indonesia; Department of Basic Nursing, Faculty of Health Sciences, Universitas Brawijaya, Malang, Indonesia
