牛蒡子苷减轻脂多糖诱导人鼻咽上皮细胞炎症损伤的作用

Effects of arctiin relieving inflammatory injury of human nasopharyngeal epithelial cells induced by lipopolysaccharide

目的探讨牛蒡子苷(ARC)减轻脂多糖(LPS)诱导的人鼻咽上皮细胞NP-69炎症损伤的作用。方法采用MTS法测定0.0001、0.001、0.01、0.1、1.0、10μmol/L ARC作用24 h对NP-69细胞增殖的影响。采用划痕实验分别检测经0.01、0.1、1.0μmol/L ARC作用24 h和经0.01、0.1、1.0μmol/L ARC预处理24 h+1.0μg/mL LPS刺激24 h后NP-69细胞的迁移情况。以LPS刺激建立NP-69细胞炎症损伤模型,检测上清液中一氧化氮(NO)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β水平以及细胞中胞质紧密连接蛋白1(ZO-1)、β-防御素3(BD3)、Janus激酶1(JAK1)、信号转导及转录活化因子3(STAT3)mRNA和蛋白的相对表达量。结果与正常组比较,0.0001、0.001、0.01、0.1、1.0、10μmol/L ARC作用24 h对NP-69细胞增殖均无影响(P>0.05)。0.1、1.0μmol/L ARC均可显著提高细胞的迁移率(P<0.05)。对LPS刺激NP-69细胞造成的炎症损伤,ARC(1.0μmol/L)能显著减少NO、TNF-α、IL-6的释放(P<0.05),同时能显著上调ZO-1、BD3 m RNA和蛋白的表达并下调STAT3 mRNA和蛋白的表达(P<0.01或P<0.05)。结论ARC具有明显减轻LPS致NP-69细胞炎症损伤的作用,并可增强炎症环境下鼻黏膜上皮屏障的物理及免疫防御功能;上述作用可能与抑制IL-6/JAK1/STAT3信号通路相关。

OBJECTIVE To investigate the effect of arctiin(ARC)relieving lipopolysaccharide(LPS)induced inflammatory injury of human nasopharyngeal epithelial cells NP-69. METHODS The effects of 24 h treatment of 0.000 1,0.001,0.01,0.1,1.0,10 μmol/L ARC on the proliferation of NP-69 were determined by MTS method. After 0.01,0.1,and 1.0 μmol/L ARC was applied to NP-69 for 24 h and NP-69 was pre-treated with 0.01,0.1 and 1.0 μmol/L ARC for 24 h,and then stimulated with 1.0μg/mL LPS for 24 h,scratch tests were used to detect cell migration in both experiments. LPS stimulated NP-69 to establish an inflammation injury model. The levels of nitric oxide(NO),tumor necrosis factor α(TNF-α)interleukin-6(IL-6),and IL-1β in cell supernatants were detected,and m RNA and protein expression of zonula oecludens protein 1(ZO-1),β-defensin 3(BD3),Janus kinase 1(JAK1),signal transducer and activator of transcription 3(STAT3) in cell supernatant were also detected.RESULTS Compared with normal group,0.000 1,0.001,0.01,0.1,1.0,10 μmol/L ARC had no effect on the proliferation of NP-69 after 24 h treatment(P>0.05). ARC(0.1,1.0 μmol/L)could significantly promote the rate of cell migration(P<0.05).For the inflammatory injure of NP-69 cells stimulated by LPS,ARC(1.0 μmol/L)could significantly reduce the release of NO,TNF-α and IL-6(P<0.05),significantly increased mRNA and protein expression of ZO-1 and BD3 but decreased mRNA and protein expression of STAT3(P<0.01 or P<0.05). CONCLUSIONS ARC has the effect of reducing the inflammatory injury of NP-69 cells induced by LPS,promoting the physical and immune defense ability of the nasal mucosa epithelial barrierunder inflammatory environment. The mechanism of action may be related to inhibiting IL-6/JAK1/STAT3 signaling pathway.