Study of MMP-9 gene expression in the presence of Sialic acid in human Glialcell line

Introduction: Sialic acid (N-acetylneuraminic acid, NANA) is found at all cell surfaces of vertebrates which can cause many different reactions, such as starting or inhibiting the immune response, the activating of the complement system and cell signaling. Sialic acid involvement in the signaling pathways leading to MMP-9 expression in Glial cells is unclear. Therefore, this study investigated the relationship between MMP-9 and NANA and the effects of these ligand on the signaling processes of the inflammatory demyelination. Materials and methods: Human Glial cell line was prepared from Pasteur Institute of Iran and cultured in DMEM, supplemented with 10% FBS. The IC50 value of NANA was obtained by MTT assay. Glial cell line was treated with NANA (300,500 and 1000 μg/ml) for 24h to investigate the effects of these ligand on the expression of MMP-9. Then total cellular RNA was isolated from approximately 6-5×106 Glial cells. 1000 ng of total RNA from each sample was used into cDNA. Real-time PCR determined the expression level of the MMP-9 transcripts and REST and SPSS software were used for analyze. Results: The results of MTT assay were analyzed by Excel software and IC50 was obtained equivalent to 1273.3 μM. Therefore, the concentration of Sialic acid was selected for treatments that were less than Ic50 (300,500 and 1000 μM). By analyzing Real-time data, it was found that MMP-9mRNA expression was up-regulated with treatment and indicated a possible involvement NANA on signaling pathway this gene expression. Conclusion: By determining the possible relationship between NANA and MMP-9 gene expression, which is one of a key inflammation factors in some neurodegenerative diseases, should be investigated MMPs gene control agents (including TIMPs and miRNAs); as well as other inflammatory macromolecules such as cytokines, chemokines, prostaglandins, NO and ROS.

Keywords:

Sialic acid (N-acetylneuraminic acid) , NANA , MMP-9 , Glial cell line , Multiple Sclerosis (MS)