Investigation of the Inhibitory Mechanisms of Resveratrol on Mycoplasma fermentans-Derived Lipoprotein MALP-2-Induced Matrix Metalloproteinase-9 Activation in THP-1 Cells

• Main Authors: Yu-Hsuan Chiang, 江妤璇
• Other Authors: George Hsiao
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/80746373846425672541

碩士 === 臺北醫學大學 === 藥理學研究所 === 94 === Infection with bacteria have been reported to be associated with human inflammation reaction. Typicallys macrophages and monocytes are activated by components of the microbial cell wall such as peptidoglycan fragments, LPS, lipoteichoic acid, and bacterial lipoprotein. Surprisingly, cell wall-less mycoplasmas can also activated mcarophages very efficiently. Mycoplasma lipoproteins have been demonstrated to activated monocytic cells and induce proinflammatory chtokine secretion. Accumulating evidence indicates that human monocytes and macrophages synthesize and secrete a family of zinc-dependent neutral endopeptidases named matrix metalloproteinases (MMPs), which play an important role in matrix remodeling, repairing and destroying. Resveratrol, a naturally occurring polyphenolic phytoalexin, is found in a wide variety of plants, which exhibits a number of biological activites, including anti-inflammatory and anticarcinogenic properties. We used Mycoplasma fermentans-derived macrophage-activating lipopeptide (MALP-2) as a stimulus that induced expression of MMP-9 in human monocytic THP-1 cells. According to gelatin zymography, we found that the expression and activation of MMP-9 protein induced by MALP-2 were inhibited by resveratrol in a concentration-dependent way. We also found that the inhibitory effect of resveratrol was not due to an impairment of cellular viability as measured by MTT assays. According to Western blot analysis, we observed that the inhibition on MALP-2-induced expression of MMP-9 protein by resveratrol was concentration-dependent. Furthermore, we found that at higher concentration (20micro Molar), resveratrol significantly reduced TIMP-1 proteins measured by enzyme-linked immunosorbent assay (ELISA). In the transcription level, resveratrol also suppressed the MALP-2-induced MMP-9 mRNA expression measured by RT-PCR. We found that resveratrol significantly inhibited the degradation of total inhibitor-kappaB-alpha (IkappaBalpha) and suppressed Akt expression induced by MALP-2in Western blot analysis. Regarding MAPKs, resveratrol suppressed extracellular signal-regulated kinase 1 and 2 (ERK1/2) expression in MALP-2 stimulation. In summary, we found that resveratrol had inhibitory effect on MMP-9 expression and on activation in THP-1 cells. Its main mechanism of action might be through NF-kappaB and MAPK signal pathway on MALP-2stimulation. It is interesting to do further studies and investigations on resveratrol as therapeutic targets in inflammatory and cancer research in vivo.