Elham Moshtaghie - Department of Biochemistry, Islamic Azad University, Falavarjan Branch, Isfahan, Iran
Hashem Naieri - Department of Biochemistry, Islamic Azad University, Falavarjan Branch, Isfahan, Iran
Ali Asghar Moshtaghie - Department of Biochemistry, Islamic Azad University, Falavarjan Branch, Isfahan, Iran
Sedigheh Asgary - Department of Biochemistry, Islamic Azad University, Falavarjan Branch AND Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, Iran

BACKGROUND: The important role of lipoproteins, particularly low-density lipoprotein (LDL) and high-density lipoprotein (HDL), has been highly regarded among the known causes of cardiovascular disease (CVD). A wide range of risk factors may cause structural and functional changes in lipoprotein particles, resulting in deposition and formation of atherosclerotic plaques. Homocysteine is one of the most important risk factors in heart disease, and its atherosclerotic properties appear to be related to its intermediate metabolite called homocysteine thiolactone (HCTL). The major aim of the present investigation was to study the effect of HCTL in different concentrations (۱۰, ۵۰, and ۱۰۰ μM) on paraoxonase and aryl esterase activities of purified human serum paraoxonase ۱ (PON۱) antioxidant enzyme related to HDL, as an extracellular hydrolyzing enzyme of HCTL.METHODS: In order to purify PON۱ enzyme from human serum, three-step chromatographic methods including DEAE Sephadex A۵۰, Sephadex G۱۰۰, and DEAE Sephadex A۵۰ were used. Protein concentration and paraoxonase and aryl esterase activities of each fraction were measured separately and the highest activities fractions were collected and subsequently pooled together for the next steps. Ultimately, both activities of PON۱ in the presence of different concentrations of HCTL were measured in triplicate by spectrophotometry technique.RESULTS: HCTL at concentrations of ۵۰ and ۱۰۰ μM decreased both paraoxonase and aryl esterase activities (P < ۰.۰۵) in comparison with the control group, which is directly related to the increase in HCTL concentration. However, at a concentration of ۱۰ μM HCTL, no significant difference was observed in both paraoxonase and aryl esterase activities compared to the control group.CONCLUSION: HCTL is a highly toxic and reactive compound that is produced in all cells. Extracellular enzyme PON۱ causes its hydrolysis with high efficiency. The results obtained from the present study showed that paraoxonase and aryl esterase activities decreased in vitro in the presence of HCTL and therefore, HCTL may cause changing in the protein structure of this enzyme. Previous in vivo studies have also shown decrease of PON۱ activity in patients with hyperhomocysteinemia.

Atherosclerosis, Homocysteine Thiolactone, Paraoxonase ۱