摘要： 目的 探讨黄芪甲苷（AS-Ⅳ）对脂多糖（LPS）诱导的巨噬细胞RAW264.7炎症的保护作用及对核因子κB受体活化因子配体（RANKL）/骨保护素（OPG）系统表达的影响。方法 将LPS刺激的RAW264.7细胞作为模型组，CCK-8法检测细胞活力，ELISA法检测白细胞介素-1β （IL-1β）、白细胞介素-6 （IL-6）和肿瘤坏死因子-α （TNF-α）炎性细胞因子的表达，实时PCR法检测RANKL与OPG的mRNA表达，Western blotting法检测Toll样受体4 （TLR4）、磷酸化核因子κB抑制剂α （P-IκBα）、磷酸化核因子κB （P-NF-κB P65）、RANKL和OPG蛋白的表达。结果 CCK-8法显示AS-Ⅳ药物浓度0~100 μg/mL时，其细胞活力无统计学差异，在LPS的诱导下，50 μg/mL、100 μg/mL的AS-Ⅳ对细胞活性有明显的提高作用；AS-Ⅳ使炎性细胞因子水平显著降低，显著下调TLR4、P-IκBα和P-NF-κB P65蛋白的表达；同时AS-Ⅳ下调了RANKL蛋白与mRNA的表达，上调了OPG蛋白与mRNA的表达。结论 AS-Ⅳ可通过调节NF-κB信号通路抑制炎症作用，并下调RANKL蛋白与mRNA的表达，上调OPG蛋白与mRNA的表达，从而调控骨代谢。

黄芪甲苷对脂多糖诱导的巨噬细胞炎症反应及核因子κB受体活化因子配体/骨保护素系统表达的影响

邵帅¹, 鲁美丽², 高秀秋¹, 王洪新²

1. 锦州医科大学附属第二医院牙周科, 辽宁 锦州 121000;
2. 锦州医科大学药学院药理教研室, 辽宁 锦州 121000

收稿日期:2021-07-19出版日期:2022-04-20发布日期:2022-05-05
通讯作者:王洪新E-mail:jyhxwang@163.com
作者简介:邵帅(1994-),男,硕士研究生.
基金资助:国家自然科学基金（81973553）；辽西地区口腔微创临床治疗中心创新平台（201820102）


关键词: 黄芪甲苷, 脂多糖, 炎症, 核因子κB受体活化因子配体, 骨保护素
Abstract: Objective To investigate the protective effect of astragaloside Ⅳ (AS-Ⅳ) on lipopolysaccharide (LPS) -induced RAW264.7 cell inflammation and its effect on receptor activator of nuclear factor-kappa B ligand/osteoprotegerin (RANKL/OPG) signaling. Methods LPS-stimulated RAW264.7 macrophage cells were used as the model group. Cell viability was detected by the CCK-8 assay. Expressions of interleukin (IL)-1β,IL-6,and tumor necrosis factor-alpha (TNF-α) inflammatory cytokines were detected by ELISA. Expressions of RANKL and OPG mRNA were detected by quantitative RT-PCR. Toll-like receptor 4 (TLR4),phosphonated inhibitor of kappa-B (P-I κBα),phosphonated NF-κB p65,RANKL,and OPG protein expressions were detected by Western blotting. Results In the presence of LPS,50 and 100 μg/mL astragaloside Ⅳ significantly increased cell activity. Astragaloside Ⅳ significantly reduced the levels of inflammatory factors,and significantly down-regulated the protein expressions of TLR4,P-IκBα,and P-NF-κB p65. Astragaloside Ⅳ also down-regulated the expressions of RANKL protein and mRNA,and up-regulated the expressions of OPG protein and mRNA. Conclusion Astragaloside Ⅳ can inhibit inflammation by regulating the NF-κB signaling pathway,down-regulating the expression of RANKL protein and mRNA,and up-regulating the expression of OPG protein and mRNA. These activities inhibit the formation of osteoclasts and improve bone repair.
Key words: astragaloside Ⅳ, lipopolysaccharide, inflammation, receptor activator of nuclear factor-κB ligand, osteoclastogenesis inhibitory factor
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