首页> 外文期刊>Cellular Physiology and Biochemistry >β-Arrestin2 Inhibits Expression of Inflammatory Cytokines in BEAS-2B Lung Epithelial Cells Treated with Cigarette Smoke Condensate via Inhibition of Autophagy
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β-Arrestin2 Inhibits Expression of Inflammatory Cytokines in BEAS-2B Lung Epithelial Cells Treated with Cigarette Smoke Condensate via Inhibition of Autophagy

机译:β-Arrestin2通过抑制自噬抑制香烟烟雾冷凝物处理后的BEAS-2B肺上皮细胞中炎性细胞因子的表达

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Background/Aims β-arrestin2 has been shown to have a role in human inflammatory disease. However, the role of β-arrestin2 in cigarette smoke-induced inflammation in the lung remains unknown. The aims of this study were to investigate the effects of β-arrestin2 on cigarette smoke condensate (CSC)-induced expression of inflammatory cytokines in the BEAS-2B human bronchial epithelial cell line in vitro, and the mechanisms involved. Methods The MTT assay determined cell viability of cultured BEAS-2B cells. Autophagy was assessed by western blot, adenoviral mRFP-GFP-LC3 transfection, and immunofluorescence. The effects of β-arrestin2 shRNA knockdown were studied by western blot and real-time reverse transcription-polymerase chain reaction (RT-PCR). Western blot evaluated the AMPK/mTOR signaling pathway. Levels of inflammatory cytokines, interleukin (IL)-6, IL-8, and MCP-1 were measured in cell culture supernatants by enzyme-linked immunosorbent assay (ELISA). Results CSC suppressed expression of β-arrestin2 in BEAS-2B cells, activated the AMPK/mTOR signaling pathway, increased cell autophagy and the expression of IL-6, IL-8, and MCP-1,pretreatment with the β-arrestin2 biased ligands, propranolol, and ICI118551 reversed these changes. Inhibition of autophagy reduced the expression of inflammatory cytokines following CSC. Conclusion In the human bronchial epithelial cell line, BEAS-2B, β-arrestin2 reduced the expression of CSC-induced inflammatory cytokines by inhibiting autophagy, most likely via the AMPK/mTOR signaling pathway.
机译:背景/目的已显示β-arrestin2在人类炎性疾病中起作用。然而,β-arrestin2在香烟烟雾引起的肺部炎症中的作用仍然未知。这项研究的目的是研究β-arrestin2对香烟烟雾冷凝物(CSC)诱导的BEAS-2B人支气管上皮细胞系中炎性细胞因子表达的影响及其机制。方法MTT法测定培养的BEAS-2B细胞的细胞活力。通过蛋白质印迹,腺病毒mRFP-GFP-LC3转染和免疫荧光评估自噬。通过蛋白质印迹和实时逆转录聚合酶链反应(RT-PCR)研究了β-arrestin2shRNA敲低的作用。 Western blot评估了AMPK / mTOR信号通路。通过酶联免疫吸附测定(ELISA)测定细胞培养上清液中的炎症细胞因子,白介素(IL)-6,IL-8和MCP-1的水平。结果CSC抑制了BEAS-2B细胞中β-arrestin2的表达,激活了AMPK / mTOR信号通路,增强了细胞的自噬作用,并表达了IL- 6,IL-8和MCP-1。 ,心得安和ICI118551扭转了这些变化。自噬的抑制降低了CSC后炎性细胞因子的表达。结论在人支气管上皮细胞系BEAS-2B中,β-arrestin2通过抑制自噬降低了CSC诱导的炎性细胞因子的表达,这很可能是通过AMPK / mTOR信号通路。

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