首页> 外文期刊>Clinical and experimental allergy : >Platelet-derived growth factor and transforming growth factor-beta modulate the expression of matrix metalloproteinases and migratory function of human airway smooth muscle cells.
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Platelet-derived growth factor and transforming growth factor-beta modulate the expression of matrix metalloproteinases and migratory function of human airway smooth muscle cells.

机译:血小板衍生的生长因子和转化生长因子-β调节基质金属蛋白酶的表达和人类气道平滑肌细胞的迁移功能。

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BACKGROUND: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMPs) have been suggested to be involved in the pathogenesis of asthma. Their expression in airway smooth muscle (ASM) cells could be involved in collagen turnover and migration of these cells and thus may contribute to airway remodelling. OBJECTIVE: To examine the effect of pro-fibrotic growth factors TGF-beta and platelet-derived growth factor (PDGF) on the expression of MMPs/TIMPs in cultured human ASM cells and to examine the role of MMP in the migration of ASM cells. METHODS: ASM cells were stimulated with TGF-beta and/or PDGF. Expression and activity of MMP-1, MMP-2, MMP-3, TIMP-1 and TIMP-2 were evaluated by quantitative RT-PCR, Western blot and zymography. Modified Boyden-chamber migration assay was performed to investigate the effect of secreted MMP-3 and TIMP-1 on ASM-cell migration. RESULTS: PDGF strongly up-regulated the expression of MMP-1 at mRNA and protein levels. PDGF, when combined with TGF-beta, caused synergistic up-regulation of MMP-3. TIMP-1 was additively up-regulated by TGF-beta and PDGF. These growth factors had no effect on the expression of MMP-2 and TIMP-2. U0126, an extracellular signal-regulated kinase (ERK) pathway inhibitor, inhibited the up-regulation of MMP-1 by PDGF. The synergistic/additive up-regulation of MMP-3 and TIMP-1 was inhibited by U0126 and SB431542, a Smad pathway inhibitor. Supernatant from ASM cells in which MMP-3 production was knocked down by RNA interference showed a decreased migratory effect on ASM cells, whereas supernatant from cells with suppressed TIMP-1 expression resulted in increased migration. CONCLUSION: Our results suggest that PDGF with/without TGF-beta could facilitate migration of ASM cells by modification of MMP-TIMP balance through the ERK pathway.
机译:背景:基质金属蛋白酶(MMPs)和金属蛋白酶组织抑制剂(TIMPs)已被认为与哮喘的发病机制有关。它们在气道平滑肌(ASM)细胞中的表达可能与这些细胞的胶原蛋白更新和迁移有关,因此可能有助于气道重塑。目的:探讨促纤维化生长因子TGF-β和血小板源性生长因子(PDGF)对培养的人ASM细胞中MMPs / TIMPs表达的影响,并探讨MMP在ASM细胞迁移中的作用。方法:用TGF-β和/或PDGF刺激ASM细胞。通过定量RT-PCR,Western印迹和酶谱法评估MMP-1,MMP-2,MMP-3,TIMP-1和TIMP-2的表达和活性。进行了改良的博登室迁移试验,以研究分泌的MMP-3和TIMP-1对ASM细胞迁移的影响。结果:PDGF在mRNA和蛋白质水平上强烈上调了MMP-1的表达。 PDGF与TGF-beta结合使用时,会引起MMP-3的协同上调。 TIMP-1被TGF-beta和PDGF加成上调。这些生长因子对MMP-2和TIMP-2的表达没有影响。 U0126是一种细胞外信号调节激酶(ERK)途径抑制剂,可抑制PDGF对MMP-1的上调。 U0126和Smad途径抑制剂SB431542抑制MMP-3和TIMP-1的协同/加性上调。 RNA干扰导致MMP-3产生被抑制的ASM细胞上清液对ASM细胞的迁移效果降低,而TIMP-1表达受到抑制的细胞上清液则导致迁移增加。结论:我们的结果表明,有/无TGF-β的PDGF可以通过改变ERP途径的MMP-TIMP平衡来促进ASM细胞的迁移。

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