Department of Dentistry, Faculty of Medicine and Dentistry, Katz Group Center for Pharmacy and Health Research, University of Alberta, Edmonton, Alberta.
J Periodontol. 2017 Jul;88(7):681-692. doi: 10.1902/jop.2017.160464. Epub 2017 Mar 3.
Salmeterol is a long-acting β2-adrenergic receptor agonist used to treat chronic obstructive pulmonary disease. The authors of the current study previously showed that preincubation of primary microglial-enriched cells with salmeterol could inhibit the inflammatory response induced by Escherichia coli lipopolysaccharide (LPS), a Toll-like receptor (TLR)-4 agonist. In this study, the authors sought to determine if salmeterol had a similar inhibitory effect on the inflammatory response of the murine macrophage cell line RAW264.7 and human monocyte THP-1 to LPS from Porphyromonas gingivalis (PgLPS), an oral microbe implicated in the pathogenesis of periodontal disease.
RAW264.7 and THP-1 cells were pretreated with salmeterol, followed by PgLPS, and monitored for production of inflammatory mediators by enzyme-linked immunosorbent assay. The nitric oxide concentration and nuclear factor-kappa B (NF-κB) activity were measured by Griess method and secretory alkaline phosphatase reporter activity assay, respectively. Reverse-transcriptase polymerase chain reaction and immunoblot analysis were used to measure messenger RNA and protein levels. Nuclear translocation of NF-κB was detected by immunofluorescence.
Pretreatment with salmeterol significantly inhibited production of proinflammatory mediators by RAW264.7 and THP-1 cells. Salmeterol downregulated PgLPS-mediated phosphorylation of the extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinase but not p38 mitogen-activated protein kinases (MAPKs). Salmeterol also attenuated activation of NF-κB via inhibition of nuclear translocation of p65-NFκB, the transcriptional activity of NF-κB and IκBα phosphorylation.
Salmeterol can significantly inhibit activation of macrophage-mediated inflammation by PgLPS, suggesting that use of salmeterol may be an effective treatment in inhibiting or lessening the inflammatory response mediated through TLR pathway activation.
沙美特罗是一种长效β2 肾上腺素能受体激动剂，用于治疗慢性阻塞性肺疾病。本研究的作者先前表明，用沙美特罗预先孵育原代小胶质细胞富集细胞可抑制大肠杆菌脂多糖（LPS），即 Toll 样受体（TLR）-4 激动剂诱导的炎症反应。在这项研究中，作者试图确定沙美特罗是否对 LPS 诱导的鼠巨噬细胞系 RAW264.7 和人单核细胞 THP-1 的炎症反应具有类似的抑制作用，LPS 来自牙龈卟啉单胞菌（PgLPS），一种与牙周病发病机制有关的口腔微生物。
用沙美特罗预处理 RAW264.7 和 THP-1 细胞，然后用 PgLPS 处理，并通过酶联免疫吸附试验监测炎症介质的产生。通过 Griess 法和分泌碱性磷酸酶报告活性测定分别测量一氧化氮浓度和核因子-κB（NF-κB）活性。逆转录聚合酶链反应和免疫印迹分析用于测量信使 RNA 和蛋白水平。通过免疫荧光检测 NF-κB 的核易位。
沙美特罗预处理显著抑制 RAW264.7 和 THP-1 细胞促炎介质的产生。沙美特罗下调了 PgLPS 介导的细胞外信号调节激酶 1/2 和 c-Jun N 末端激酶的磷酸化，但不影响 p38 丝裂原活化蛋白激酶（MAPKs）。沙美特罗还通过抑制 p65-NFκB 的核易位、NF-κB 的转录活性和 IκBα 磷酸化来减弱 NF-κB 的激活。
沙美特罗可显著抑制 PgLPS 激活的巨噬细胞介导的炎症反应，表明使用沙美特罗可能是抑制或减轻通过 TLR 途径激活介导的炎症反应的有效治疗方法。
