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The study used the AirQ + software developed by the World Health Organization (WHO) to evaluate the health impacts associated with long-term exposure to PM2.5 in Mashhad, Iran. For this purpose, we analyzed the daily average concentrations of PM2.5 (with a diameter of 2.5 micrometers or less) registered by the air quality monitoring stations from 2016 to 2021. The levels of PM2.5 surpassed the Air Quality Guidelines (AQG) limit value of 5 µg/m3 (annual value) established by WHO. The findings revealed that the burden of mortality (from all-natural causes) at people above 30 years old associated with PM2.5 exposures was 2093 [95% confidence interval [CI]: 1627–2314] deaths in 2016 and 2750 [95% CI: 2139–3038] deaths in 2021. In general, the attributable mortality from specific causes of deaths (e.g., COPD (chronic obstructive pulmonary diseases), IHD (ischemic heart diseases) and stroke) in people above 25 years old increased between the years, but the mortality from lung cancer was stable at 46 [95% CI: 33–59] deaths in 2016 and 48 [95% CI: 34–61] deaths in 2021. The attributable mortality from ALRI (Acute Lower Respiratory Infection) in children below 5 years old increased between the years. We also found differences in mortality cases from IHD and stroke among the age groups and between the years 2016 and 2021. It was concluded that burden of disease methodologies are suitable tools for regional and national policymakers, who must take decisions to prevent and to control air pollution and to analyze the cost-effectiveness of interventions. [ABSTRACT FROM AUTHOR]