Isorhamnetin attenuates TNF-α-induced inflammation, proliferation, and migration in human bronchial epithelial cells via MAPK and NF-κB pathways
Isorhamnetin attenuates TNF-α-induced inflammation, proliferation, and migration in human bronchial epithelial cells via MAPK and NF-κB pathways
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异鼠李素通过 MAPK 和 NF-κB 途径减弱 TNFαα 诱导的人支气管上皮细胞炎症、增殖和迁移
DOI：
10.1002/ar.24506
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发表时间：
2020-09-24
影响因子：
2
通讯作者：
Wang, Yong
中科院分区：
文献类型：
--
作者：
Ren, Xiaojie;Han, Longyin;Wang, Yong
异鼠李素具有独特的抗炎活性并抑制细胞增殖和迁移。这些影响也与哮喘的发病机制有关。然而，异鼠李素对哮喘患者支气管上皮细胞的影响尚未得到研究。将人支气管上皮细胞系BEAS-2B的细胞与异鼠李素和肿瘤坏死因子(TNF)-α一起培养。使用 CCK8 测定评估异鼠李素对 BEAS-2B 细胞活力的影响。进行 EdU（5-乙炔基-2'-脱氧尿苷）细胞增殖测定来评估细胞增殖。使用 Transwell 和伤口愈合测定来测量 BEAS-2B 细胞迁移。进行实时PCR和酶联免疫吸附测定来测量促炎细胞因子的表达。通过蛋白质印迹法测定蛋白质表达水平。免疫荧光用于检测核因子κB（NF-κB）的核转位。我们发现20和40μM的异鼠李素减少了TNF-α诱导的BEAS-2B细胞的增殖。异鼠李素显着降低 TNF-α 诱导的 BEAS-2B 细胞中白介素 (IL)-1 beta、IL-6、IL-8 和 C-X-C 基序趋化因子配体 10 的表达。此外，10μM异鼠李素有效减少TNF-α诱导的细胞迁移。异鼠李素治疗可抑制 TNF-α 诱导的丝裂原激活蛋白激酶 (MAPK) 和 NF-κ B 通路的磷酸化。综上所述，异鼠李素通过调节 MAPK 和 NF-κ B 信号通路抑制 BEAS-2B 细胞的炎症、增殖和迁移，是治疗哮喘的候选药物。
Isorhamnetin has distinct anti-inflammatory activity and inhibits cell proliferation and migration. These effects are also involved in the pathogenesis of asthma. However, the effect of isorhamnetin on bronchial epithelial cells in patients with asthma has not been examined. Cells of human bronchial epithelial cell line BEAS-2B were cultured with isorhamnetin and tumor necrosis factor (TNF)-alpha. The effects of isorhamnetin on BEAS-2B cell viability were assessed using CCK8 assay. The EdU (5-ethynyl-2 '-deoxyuridine) cell proliferation assay was performed to assess cell proliferation. BEAS-2B cell migration was measured using Transwell and wound healing assays. Real-time PCR and enzyme-linked immunosorbent assay were conducted to measure the expression of pro-inflammatory cytokines. Protein expression levels were determined by western blotting. Immunofluorescence was used to detect nuclear translocation of nuclear factor kappa B (NF-kappa B). We found that isorhamnetin at 20 and 40 mu M reduced the proliferation of BEAS-2B cells induced by TNF-alpha. Isorhamnetin significantly decreased the expression of interleukin (IL)-1 beta, IL-6, IL-8, and C-X-C motif chemokine ligand 10 in BEAS-2B cells induced by TNF-alpha. Additionally, 10 mu M isorhamnetin effectively reduced cell migration induced by TNF-alpha. Treatment with isorhamnetin inhibited the phosphorylation of mitogen-activated protein kinase (MAPK) and NF-kappa B pathways induced by TNF-alpha. In summary, isorhamnetin inhibited the inflammation, proliferation, and migration of BEAS-2B cells by regulating the MAPK and NF-kappa B signaling pathways and is a drug candidate for asthma.