Regulation of matrix metalloproteinase-3 gene expression in inflammation: A molecular study.

Abstract

Introduction:Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways.Materials and Methods:Lipopolysaccharide-induced murine macrophage-like RAW 264.7 cell lines were obtained and maintained in Dulbecco's modified Eagle's medium plus 10% fetal bovine serum under optimal temperatures. Primers used were MMP-3 forward primer, MMP-3 reverse primer, and glyceraldehyde-3-phosphate dehydrogenase forward primer and glyceraldehyde-3-phosphate reverse primer. Total RNA was isolated, the sample was prepared, and electrophoresis was performed. The first strand of cDNA was synthesized and amplification of specific isolated gene using polymerase chain reactor (PCR). The amplified products were then separated on a 1.0% agarose gel in 1XTBE at 75 V for 3 h. The gel was stained with ethidium bromide, and the amplified product was visualized and photographed on Gel Doc system.Results:Real-time PCR showed only bands at expected size of 595 bp for internal control amplification of glyceraldehyde-3-dehydrogenase gene. Analysis was done with densitometry, and these values are compared with the negative control. Results showed a statistically significant rise in the relative levels of MMP-3-mRNA when compared with negative control at 1, 2, and 3 h.Conclusion:This study proved the significantly increased levels of MMP gene at different period, thereby it can be concluded that MMP-3 levels are higher in inflammatory conditions.