Mechanical stress-induced interleukin-1beta expression through adenosine triphosphate/P2X7 receptor activation in human periodontal ligament cells
Mechanical stress-induced interleukin-1beta expression through adenosine triphosphate/P2X7 receptor activation in human periodontal ligament cells
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DOI：
10.1111/j.1600-0765.2012.01517.x
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发表时间：
2013-04-01
影响因子：
3.5
通讯作者：
Pavasant, P.
中科院分区：
文献类型：
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作者：
Kanjanamekanant, K.;Luckprom, P.;Pavasant, P.
背景与目的机械应力是维持牙周组织内环境稳定的重要因素。白细胞介素-1 β（IL-1）和三磷酸腺苷（ATP）被认为是有效的炎症介质。在巨噬细胞中，ATP激活的P2 X7受体参与IL-1的加工和释放。我们以前的工作表明，机械应力诱导的表达骨桥蛋白和RANKL通过ATP/P2 Y1受体在人牙周膜（HPDL）细胞。本研究旨在探讨机械应力对HPDL细胞IL-1表达的影响，以及ATP和P2嘌呤能受体的作用机制和参与。材料与方法体外培养的HPDL细胞，施加持续的压力负荷。分别采用RT-PCR和ELISA法在mRNA和蛋白质水平上分析IL-1的表达。使用MTT测定法检查细胞活力。ATP也被用于刺激HPDL细胞。利用抑制剂、拮抗剂和小干扰RNA（siRNA）技术研究ATP和负责IL-1诱导的特异性P2亚型的作用以及细胞内机制沿着。结果机械应力可通过ATP的释放上调HPDL细胞IL-1的表达。ATP单独也能够增加IL-1表达。IL-1的诱导被抑制剂和靶向P2 X7受体的siRNA显著抑制。ATP刺激的IL-1表达也减少了细胞内钙抑制剂。结论HPDL细胞对机械刺激具有直接反应的能力。结果表明，ATP/P2嘌呤能受体，以及细胞内钙信号，在机械应力诱导的炎症通过上调促炎细胞因子，IL-1，在HPDL细胞中的重要作用。
Background and Objective Mechanical stress is an important factor in maintaining homeostasis of the periodontium. Interleukin-1beta (IL-1) and adenosine triphosphate (ATP) are considered potent inflammatory mediators. In macrophages, ATP-activated P2X7 receptor is involved in IL-1 processing and release. Our previous works demonstrated mechanical stress-induced expression of osteopontin and RANKL through the ATP/P2Y1 receptor in human periodontal ligament (HPDL) cells. This study was designed to examine the effect of mechanical stress on IL-1 expression in HPDL cells, as well as the mechanism and involvement of ATP and the P2 purinergic receptor. Material and Methods Cultured HPDL cells were treated with continuous compressive loading. IL-1 expression was analyzed at both mRNA and protein levels, using RT-PCR and ELISA, respectively. Cell viability was examined using the MTT assay. ATP was also used to stimulate HPDL cells. Inhibitors, antagonists and the small interfering RNA (siRNA) technique were used to investigate the role of ATP and the specific P2 subtypes responsible for IL-1 induction along with the intracellular mechanism. Results Mechanical stress could up-regulate IL-1 expression through the release of ATP in HPDL cells. ATP alone was also capable of increasing IL-1 expression. The induction of IL-1 was markedly inhibited by inhibitors and by siRNA targeting the P2X7 receptor. ATP-stimulated IL-1 expression was also diminished by intracellular calcium inhibitors. Conclusion Our work clearly indicates the capability of HPDL cells to respond directly to mechanical stimulation. The results signified the important roles of ATP/P2 purinergic receptors, as well as intracellular calcium signaling, in mechanical stress-induced inflammation via up-regulation of the proinflammatory cytokine, IL-1, in HPDL cells.