Noise pollution is one of the hazardous factors in the environment. One of the major sources of noise pollution is urban transportation, mostly the subway system and transportation in business centers. In this study, the effects of noise pollution on blood serum proteins level of Wistar male rats were investigated. Rats were divided into 3 groups (1 control and 2 experimental, n=6). The control group was kept in standard conditions and the experimental groups were kept in Tehran Grand Bazaar and the vicinity of Sadeghiyeh metro-station (West Rose Street). After eight weeks, the blood serum proteins of each group were analyzed by electrophoresis assay. Then, the blood serum samples were investigated by Ultraviolet-Visible spectroscopy and chemometrics method (MCR-ALS). Assessment of serum proteins indicated that among 5 essential proteins in the blood, the percentage of alpha-globulins and beta-globulins displayed significant difference (p<0.05) in experimental groups. The UV-Visible spectra and chemometrics method showed that there were different species in both experimental groups as compared with the control group. Thus, the noise pollution of the areas studied led to alterations in blood proteins. Consequently, it seems crucial to take control and prevention measures for noise reduction in these areas.