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Cytokines induce MMP-3-regulated proliferation of embryonic stem cell-derived odontoblast-like cells

机译:细胞因子诱导MMP-3调节胚胎干细胞来源的成牙本质细胞样细胞的增殖

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Objectives: Matrix metalloproteinase (MMP)-3 expression increases after pulpectomy and accelerates angiogenesis in rat dental pulp by an uncharacterised mechanism. Odontoblasts, a major component of dental pulp, could represent a therapeutic target. We investigated whether MMP-3 activity is induced by cytokines and/or is associated with cell proliferation and apoptosis in embryonic stem cell-derived odontoblast-like cells. Materials and Methods: We used reverse transcriptase polymerase chain reaction, western blotting, an MMP-3 activity assay, a BrdU-cell proliferation enzyme-linked immunosorbent assay and DNA fragmentation analysis to evaluate siRNA-mediated downregulation of MMP-3 expression and activity, and any changes in the proliferative and apoptotic responses associated with this reduced expression. Results: Pro-inflammatory cytokines (interleukin-1β, tumour necrosis factor-α and interferon-γ, at relatively low concentrations) induced MMP-3 mRNA and protein expression, and increased MMP-3 activity and cell proliferation, but not apoptosis. MMP-3 silencing produced a potent and significant suppression of cytokine-induced MMP-3 expression and activity, decreased cell proliferation and increased apoptosis. These effects were rescued by application of exogenous MMP-3. Conclusions: Our results suggest that pro-inflammatory cytokines induce MMP-3-regulated cell proliferation and anti-apoptosis effects in odontoblast-like cells derived from embryonic stem cells, in addition to their well-documented destructive role in inflammation.
机译:目的:牙髓切除后基质金属蛋白酶(MMP)-3的表达增加,并通过未知机制促进大鼠牙髓中的血管生成。成牙本质细胞是牙髓的主要成分,可以代表治疗目标。我们调查了MMP-3活性是否由细胞因子诱导和/或与胚胎干细胞来源的成牙本质细胞样细胞中的细胞增殖和凋亡相关。材料和方法:我们使用逆转录酶聚合酶链反应,蛋白质印迹,MMP-3活性测定,BrdU细胞增殖酶联免疫吸附测定和DNA片段分析来评估siRNA介导的MMP-3表达和活性下调,以及与此表达降低相关的增殖和凋亡反应的任何变化。结果:促炎细胞因子(白细胞介素-1β,肿瘤坏死因子-α和干扰素-γ,相对较低的浓度)诱导MMP-3 mRNA和蛋白表达,并增加MMP-3活性和细胞增殖,但不诱导细胞凋亡。 MMP-3沉默可有效抑制细胞因子诱导的MMP-3的表达和活性,减少细胞增殖并增加细胞凋亡。这些作用通过应用外源性MMP-3得以挽救。结论:我们的研究结果表明,促炎细胞因子除了能证明其对炎症的破坏作用外,还能在源自胚胎干细胞的成牙本质细胞样细胞中诱导MMP-3调节的细胞增殖和抗凋亡作用。

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