Interleukin-1beta and tumor necrosis factor-alpha induce chemokine and matrix metalloproteinase gene expression in human colonic subepithelial myofibroblasts.
ABSTRACT Colonic subepithelial myofibroblasts may play a role in the inflammatory responses and in extracellular matrix (ECM) metabolism. In this study, we investigated the effects of interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha on chemokine (IL-8 and monocyte chemoattractant protein (MCP)-1) and ECM turnover (proliferation of subepithelial myofibroblasts, and secretion of ECM and matrix metalloproteinases (MMPs)) in colonic subepithelial myofibroblasts.
Human colonic subepithelial myofibroblasts were isolated using the method described by Mahida et al. Chemokine and MMP expressions were determined by ELISA and Northern blotting. Nuclear factor (NF)-kappaB and NF-IL6 DNA binding activities were evaluated by electrophoretic gel mobility shift assays (EMSA).
IL-1beta and TNF-alpha did not affect the proliferation of subepithelial myofibroblasts, but stimulated the secretion of types I and IV collagens weakly. Unstimulated subepithelial myofibroblasts secreted a large amount of MMP-2, but a small amount of IL-8, MCP-1 and MMP-1. IL-1beta and TNF-alpha both induced a dose- and time-dependent increase in IL-8, MCP-1 and MMP-1 secretion, and weakly stimulated MMP-2 secretion. IL-1beta and TNF-alpha both rapidly evoked NF-kappaB DNA-binding activity. The inhibition of NF-kappaB activation markedly blocked both IL-1beta- and TNF-alpha-induced IL-8 and MCP-1 mRNA expression, but did not affect MMP-1 mRNA expression.
These observations indicate that chemokine secretion and ECM metabolism are collectively regulated by the proinflammatory cytokines, IL-1beta and TNF-alpha, in colonic subepithelial myofibroblasts. Thus, colonic subepithelial myofibroblasts may play an important role in the pathophysiology of inflammation in the colon.
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ABSTRACT: Tumor necrosis factor-α, platelet-derived growth factor, matrix metalloproteinases 9 and 2 have very important roles in neointimal hyperplasia, which develops after endovascular injury. However, the relationships among the four factors in inducing neointimal hyperplasia are unclear. Here, we used a mouse model of femoral arterial transluminal wire injury, and examined neointimal hyperplasia within the 28 days that followed the injury. We confirmed that the neointima kept growing during the 28 days, and found that expression of TNF-α and PDGF mRNAs in femoral arteries peaked within 24 h after injury. However, MMP9 mRNA expression peaked 7 days, and MMP2 mRNA expression peaked 28 days after injury. Then, we administered exogenous TNF-α or PDGF to the peri-femoral artery following an injury, and found that exogenous TNF-α led to significantly more neointimal hyperplasia during the first 2 weeks, and PDGF led to increased neointimal hyperplasia during the second 2 weeks after injury. We also used the model of femoral artery injury in MMP9- or MMP2-deficient (MMP9-/- or MMP2-/-) mice. We found that neointimal hyperplasia was reduced in MMP9-/- mice during the first 2 weeks after injury, and neointimal hyperplasia was reduced in MMP2-/- mice during the second 2 weeks after injury. When TNF-α or PDGF was administered to the peri-femoral artery immediately after injury, TNF-α did not promote neointimal hyperplasia in MMP9-/- mice during the first 2 weeks after injury but did in MMP2-/- mice, and PDGF did not promote neointimal hyperplasia in MMP2-/- mice during the second 2 weeks after injury but did in MMP9-/- mice. We used an in vitro system to treat vascular smooth muscle cells (VSMCs) with TNF-α or PDGF; TNF-α induced MMP9, but not MMP2, expression at a fast reaction speed, while PDGF induced MMP2, but not MMP9, expression at a slow reaction speed. Meanwhile, TNF-α induced VSMCs migration in a MMP9-dependent manner, and PDGF induced VSMCs proliferation in a MMP2-dependent manner. Taken together, our studies elucidated the axis of TNF-α-MMP9-VSMCs migration and PDGF-MMP2-VSMCs proliferation, both of which contributed to the mechanism of neointimal hyperplasia formation.Journal of Molecular and Cellular Cardiology 01/2013; · 5.15 Impact Factor
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ABSTRACT: Objective: To investigate whether triptolide can prolong the survival of rat mesenchymal stem cells (MSCs) transfected with the mouse hyperpolarization-activated cyclic nucleotide-gated channel 4 (mHCN4) gene in the myocardium. Methods: Grafted cell survival was determined using a sex-mismatched cell transplantation model and analysis of Y chromosome-specific Sry gene expression from hearts harvested at different time points after cell transplantation. ELISA and RT-PCR were used to measure protein and mRNA levels, respectively, of nuclear factor (NF)-κB, IL-1β, IL-6 and TNF-α. Results: Donor cell numbers decreased over time. Pretreatment with triptolide improved graft survival both 24 (29.3 ± 0.9%) and 72 h (17.5 ± 1.2%) after transplantation of MSCs and resulted in a 2.5-fold increase in the total cell number 72 h after cell transplantation. The mRNA expression and protein content of NF-κB, IL-1β, IL-6 and TNF-α were significantly reduced in the triptolide-treated group compared with the control groups. In addition, triptolide downregulated Bax but upregulated Bcl-2 in the injected region. Conclusions: Transient treatment with triptolide may significantly improve the early survival of MSCs in vivo. The mechanism underlying this effect involves attenuating the inflammatory response via inhibition of the NF-κB signaling pathway. © 2014 S. Karger AG, Basel.Cardiology 02/2014; 128(2):73-85. · 2.04 Impact Factor
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ABSTRACT: Interleukin (IL)-17A and IL-17E (also known as IL-25) have been implicated in fibrosis in various tissues. However, the role of these cytokines in the development of intestinal strictures in Crohn's disease (CD) has not been explored. We investigated the levels ofIL-17A and IL-17E and their receptors in CD strictured and non-strictured gut, and the effects of IL-17A and IL-17E on CD myofibroblasts.. IL-17A was significantly overexpressed in strictured compared with non-strictured CD tissues, whereas no significant difference was found in the expression of IL-17E or IL-17A and IL-17E receptors (IL-17RC and IL-17RB, respectively) in strictured and non-strictured CD areas. Strictured CD explants released significantly higher amounts of IL-17A than non-strictured explants, whereas no difference was found as for IL-17E, IL-6, or tumor necrosis factor-alpha production. IL-17A, but not IL-17E, significantly inhibited myofibroblast migration, and also significantly upregulated matrix metalloproteinase (MMP)-3, MMP-12, tissue inhibitor of metalloproteinase-1 and collagen production by myofibroblasts from strictured CD tissues. Our results suggest that IL-17A, but not IL-17E, is pro-fibrotic in CD. Further studies are needed to clarify whether the therapeutic blockade of IL-17A through the anti-IL-17A monoclonal antibody secukinumab is able to counteract the fibrogenic process in CD.Fibrogenesis & Tissue Repair 07/2013; 6(1):13.