The Effect of Adiponectin on Matrix Metalloproteinase-9 (MMP-9) in Vascular Smooth Muscle Cells

Authors

1 M.Sc Student, Department of Biochemistry, Shahrekord University of Medical Sciences, Shahrekord, Iran

2 Assistant Professor, Department of Biochemistry and Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran

3 Associate Professor, Department of Biochemistry and Clinical Biochemistry Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran

4 Ph.D. Student, Cellular and Molecular ResearchCenter, Shahrekord University of Medical Sciences, Shahrekord, Iran

Abstract

Background & Aims: Atherosclerosis is a major cause of morbidity and mortality. Adiponectin reducesthe risk of heart disease, and matrix metalloproteinase-9 (MMP-9) is involved in the formation and development of atherosclerotic plaque. The aim of this study was the investigation of the effect of adiponectin on MMP-9 gene expression. It seems this hormone can reduce the risk of atherosclerosis by MMP-9 gene expression reduction. Methods: Human aorta smooth muscle cells were cultured in F12K media in appropriate environment and conditions, then, the cells were treated with 5 µg/ml adiponectin. After 24 and 48 hour, total RNA was extracted and corresponding cDNA was made. After drawing standard curve and determining the efficiency of the reaction, MMP-9 gene expression was measured by the SYBR Green kit and real time PCR method. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene was the reference gene. The amount of MMP-9 protein, compared to the β-actin protein as reference protein, was estimated with Western blot method. Results: adiponectin did not cause a ch ange in gene expression of MMP-9 in 24 hours, but in 48 hours reduced gene expression (-1.1, and -5.5, respectively). As a result of MMP-9 gene expression reduction, protein also reduced after 48 hours compared to β-actin protein. Conclusion: Through the reduction of MMP-9 protein and gene expression, adiponectin changes extra cellular matrix which may reduce the risk and complications of atherosclerosis

Keywords