Mast cells exert pro-inflammatory effects of relevance to the pathophyisology of tendinopathy
Mast cells exert pro-inflammatory effects of relevance to the pathophyisology of tendinopathy
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DOI：
10.1186/ar4374
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发表时间：
2013-01-01
影响因子：
4.9
通讯作者：
Scott, Alex
中科院分区：
文献类型：
--
作者：
Behzad, Hayedeh;Sharma, Aishwariya;Scott, Alex
简介：我们之前发现，与对照组相比，髌骨肌腱病变患者的肌腱活检中肥大细胞密度增加。本研究考察了肥大细胞对基本肌细胞功能的影响，包括炎症介质前列腺素E2 （PGE(2)）的产生、细胞外基质重塑和基质金属蛋白酶（MMP）基因转录以及胶原合成。方法：用已建立的人肥大细胞系（HMC-1）刺激原代人腱细胞。通过三维胶原晶格培养细胞，研究细胞外基质重塑。采用3-（4,5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺苯基）- 2h -四氮唑盐（MTS）法评估存活/增殖。通过实时定量聚合酶链反应（qPCR）检测COX-2、COL1A1、MMP1和MMP7 mRNA水平。Western blot检测Cox-2蛋白水平，免疫荧光染色检测I型前胶原蛋白水平。采用酶联免疫吸附试验（ELISA）测定PGE(2)水平。结果：肥大细胞刺激腱细胞产生高水平的COX-2和促炎介质PGE(2)，从而降低COL1A1 mRNA的表达。此外，肥大细胞降低了腱细胞产生的I型前胶原蛋白水平。转化生长因子β 1 （tgf - β 1）负责诱导细胞Cox-2和PGE(2)。肥大细胞增加了MMP1和MMP7的转录，并增加了腱细胞对三维胶原晶格的收缩，这一现象被一种泛mmp抑制剂（Batimastat）阻断。结论：我们的数据表明，肥大细胞来源的PGE(2)减少了胶原合成，增强了人腱细胞中MMPs的表达和活性。
Introduction: We have previously found an increased mast cell density in tendon biopsies from patients with patellar tendinopathy compared to controls. This study examined the influence of mast cells on basic tenocyte functions, including production of the inflammatory mediator prostaglandin E2 (PGE(2)), extracellular matrix remodeling and matrix metalloproteinase (MMP) gene transcription, and collagen synthesis.Methods: Primary human tenocytes were stimulated with an established human mast cell line (HMC-1). Extracellular matrix remodeling was studied by culturing tenocytes in a three-dimensional collagen lattice. Survival/proliferation was assessed with the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium salt (MTS) assay. Levels of mRNA for COX-2, COL1A1, MMP1, and MMP7 were determined by quantitative real-time polymerase chain reaction (qPCR). Cox-2 protein level was assessed by Western blot analysis and type I procollagen was detected by immunofluorescent staining. PGE(2) levels were determined using an enzyme-linked immunosorbent assay (ELISA).Results: Mast cells stimulated tenocytes to produce increased levels of COX-2 and the pro-inflammatory mediator PGE(2), which in turn decreased COL1A1 mRNA expression. Additionally, mast cells reduced the type I procollagen protein levels produced by tenocytes. Transforming growth factor beta 1 (TGF-beta 1) was responsible for the induction of Cox-2 and PGE(2) by tenocytes. Mast cells increased MMP1 and MMP7 transcription and increased the contraction of a three-dimensional collagen lattice by tenocytes, a phenomenon which was blocked by a pan-MMP inhibitor (Batimastat).Conclusion: Our data demonstrate that mast cell-derived PGE(2) reduces collagen synthesis and enhances expression and activities of MMPs in human tenocytes.