Rho-kinase inhibitor Y-27632 downregulates LPS-induced IL-6 and IL-8 production via blocking p38 MAPK and NF-κB pathways in human gingival fibroblasts
Rho-kinase inhibitor Y-27632 downregulates LPS-induced IL-6 and IL-8 production via blocking p38 MAPK and NF-κB pathways in human gingival fibroblasts
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Rho 激酶抑制剂 Y-27632 通过阻断人牙龈成纤维细胞中的 p38 MAPK 和 NF-kappa B 途径下调 LPS 诱导的 IL-6 和 IL-8 产生
DOI：
10.1002/jper.17-0571
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发表时间：
2018-07-01
影响因子：
4.3
通讯作者：
Ge, Shaohua
中科院分区：
文献类型：
--
作者：
Kang, Wenyan;Shang, Lingling;Ge, Shaohua
背景：牙龈卟啉单胞菌脂多糖（LPS）在慢性牙周炎的发生和发展中起重要作用。人类牙龈成纤维细胞（HGF）与细菌或细菌产物相互作用，并触发破坏牙周组织的炎症信号通路。RhoA调节各种细胞类型中的细胞因子产生。本研究探讨Rho激酶抑制剂Y-27632在LPS诱导的核因子-κ B表达中的作用。（NF-κ B）和p-38丝裂原活化蛋白激酶（MAPK）活化，以及HGF中炎性细胞因子的产生。在脂多糖（LPS）处理的HGF中评估Y-27632、SB 203580（p38 MAPK抑制剂）和BAY 11 -7082（NF-κ B抑制剂）的作用。细胞毒性试验用于确定药物对HGF活力的影响。应用酶联免疫吸附试验和实时定量聚合酶链反应检测白细胞介素（IL）-6、IL-8和Toll样受体（TLR）水平。结果：5 μ g/mL牙龈卟啉单胞菌LPS、10 μ M Y-27632、10 μ M SB 203580和5 μ M BAY 11 -7082对HGFs无毒性作用。LPS通过增加I κ B α的降解和I κ B α、p65和p38的磷酸化，并促进p65从细胞质易位到细胞核，激活NF-κ B和p38 MAPK。NF-κ B和p38 MAPK的激活诱导IL-6和IL-8在mRNA和蛋白水平上的过度产生。结论：Rho激酶抑制剂Y-27632通过阻断HGF中NF-κ B和p38 MAPK的活化，下调LPS诱导的HGF中IL-6和IL-8的产生。
Background: Porphyromonas gingivalis lipopolysaccharide (LPS) plays a major role in the initiation and progression of chronic periodontitis. Human gingival fibroblasts (HGFs) interact with bacteria or bacterial products and trigger inflammatory signaling pathways that destroy periodontal tissues. RhoA regulates cytokine production in various cell types. This study investigated the role of Rho-kinase inhibitor Y-27632 in LPS-induced nuclear factor-kappa B (NF-kappa B) and p-38 mitogen-activated protein kinase (MAPK) activation, and inflammatory cytokine production in HGFs.Methods: Effects of Y-27632, SB203580 (p38 MAPK inhibitor), and BAY11-7082 (NF-kappa B inhibitor) were assessed in lipopolysaccharide (LPS)-treated HGFs. Cytotoxicity assays were used to determine the effect of the drugs on HGF viability. Enzyme-linked immunosorbent assays and quantitative real-time polymerase chain reaction were applied to evaluate the levels of interleukin (IL)-6, IL-8, and Toll-like receptors (TLRs). NF-kappa B and p38 MAPK pathway activation was detected by western blot and immunocytochemistry.Results: P. gingivalis LPS at 5 mu g/mL, 10 mu M Y-27632, 10 mu M SB203580, and 5 mu M BAY11-7082 exhibited no toxicity in HGFs. LPS activated NF-kappa B and p38 MAPK by increasing degradation of I kappa B alpha and phosphorylation of I kappa B alpha, p65, and p38, and facilitating p65 translocation from the cytoplasm to nuclei. The activation of NF-kappa B and p38 MAPK induced overproduction of IL-6 and IL-8 at both mRNA and protein levels. However, Y-27632 attenuated LPS-induced NF-kappa B and p38 MAPK activation and inflammatory cytokine production.Conclusions: Rho-kinase inhibitor Y-27632 downregulates LPS-induced IL-6 and IL-8 production by blocking NF-kappa B and p38 MAPK activation in HGFs.