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The Express of MMP-9 in the Atherosclerotic Tissure of Aorta and Influential Factors

Author: ZhangRong
Tutor: FeiYuXing
School: Second Military Medical University
Course: Department of Cardiology
Keywords: Atherosclerosis Matrix metalloproteinase-9 Immunohistochemical staining Vascular smooth muscle cells Western blot RT-PCR Pitavastatin
CLC: R543.5
Type: Master's thesis
Year: 2011
Downloads: 98
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Abstract

Objective: arteriosclerosis occurrence and development of chronic process is complex and affected by many factors. Patients had stable disease or not is not only related to the degree of stenosis and atherosclerosis, more closely related to plaque inflammation is active. Inflammatory cells and cytokines involved in the pathological process of changes in atherosclerotic plaque. Matrix metalloproteinase-9 (Matrix MetalloProteinase-9 MMP-9) in the process of atherosclerosis formation play an important role, MMP-9 degrade collagen, weakening the strength of the fibrous cap in atherosclerotic plaque, leading to plaque instability increased, thus contributing to the pathogenesis of acute coronary syndrome. The study observed matrix metalloproteinase-9 in human atherosclerotic tissues, and to explore the insulin-like growth factor (IGF-1), tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), monocyte chemoattractant factor -1 (MCP-1), and statins affect its expression. The Methods: arteriosclerosis tissue obtained from elective open surgical specimens, pathologic examination confirmed arteriosclerosis organization. The control group arterial specimens taken from adult specimens of unnatural deaths, general and microscopic pathologic examination confirmed normal initiative to organize. All specimens 4% paraformaldehyde fixed 12h, conventional dehydration, paraffin-embedded sections. HE staining and standard immunohistochemical staining. Human vascular smooth muscle cells taken from surgical open surgical specimens, cell separation, culture, collection, cryopreservation, recovery, RT-PCR and Western blot determination of different concentrations of TNF-α-induced vascular smooth muscle cells of MMP-9 mRNA and protein expression; different concentrations of pitavastatin intervention effect on TNF-α. The immunohistochemistry results to determine: the double-blind method by two pathologist to observe each slice. The lt; judged positive basis: light microscope (× 400) randomly selected 10 non-repeated vision, according to the percentage of positive cells Rating: 25% as negative (0), 26% to 50% as weakly positive (1 points), 51% -75% as moderately positive (2 points), gt; 76% -100% strongly positive (3 points). RT-PCR to determine methods: UV lamp observation camera, internal reference and target gene band appears to be positive. Appears only to internal control were negative. Grayscale scan scanner, the Image Tool Version2.0 software analysis. Western blot analysis of the judgment: scanner grayscale scanning Image Tool Version2.0 software analysis to detect proteins and β-actin scan grayscale ratio as an indicator of the evaluation of the protein expression levels. Results: normal abdominal aorta wall, no significant expression of MMP-9, MCP-1, IL-1β distribution, TNF-α, IGF-1 staining was positive or weakly positive; the arteriosclerosis organization MMP-9 is mainly distributed in the arterial wall in film cell infiltration areas outside the meningitis, flaky or continuous distribution, MCP-1 appears in the disordered arrangement of inflammatory cells gathered at the positive cells as fibroblasts, macrophages; IGF-1 in atherosclerotic tissue in stained weakly positive or negative, light brownish yellow positive staining occasionally in the scattered distribution of vascular smooth muscle cells. Strong expression of IL-1β, TNF-α in the normal aorta, sometimes light brown yellow staining distribution in vascular smooth muscle cells in atherosclerotic tissue distribution are mainly located in the vascular endothelial cells and vascular smooth muscle cell hyperplasia. TNF-α distribution is mainly located in the vascular membrane, outer membrane, and vessel walls in macrophages and other inflammatory cells infiltration area. MMP-9 distribution and MCP-1, TNF-α, IL-1β distribution was positively correlated with IGF-1 protein expression was negatively correlated; distribution of significant or very significant differences. TNF-α in a concentration-dependent manner (1/10/100ng/ml) increased the expression of MMP-9 mRNA and protein of vascular smooth muscle cells, the statistically significant difference (P lt; 0.01). Different concentrations of pitavastatin (0.1,1,10,100,500 nm) could antagonize TNF-α (100ng/ml) promote vascular smooth muscle cells, MMP-9 mRNA and protein expression, the difference was significant statistical significance (P lt; 0.01 ). Conclusion: arteriosclerosis organizations in both MMP-9, TNF-α, MCP-1 (IGF-1), the expression of IL-1β, but the distribution is different; TNF-α increased MMP-9 expression in vascular smooth muscle cells, pitavastatin statins could be antagonized by TNF-α on vascular smooth muscle cells MMP-9 expression. The results suggest that the interaction between the different cytokines and imbalance may be the pathological basis of unstable atherosclerotic plaque, statins by regulating the interactions between cytokines play one of the mechanisms of its anti-atherogenic effect.

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CLC: > Medicine, health > Internal Medicine > Heart, blood vessels ( circulatory ) disease > Vascular disease > Artery disease
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