麦冬皂苷D预处理对LPS介导的人肺上皮Beas-2B细胞炎性及氧化损伤的抑制作用

Inhibitory effect of ophiopogonin D pretreatment on LPS-mediated inflammatory and oxidative damage of human pulmonary epithelial Beas-2B cells

目的探讨麦冬皂苷D预处理对脂多糖(LPS)介导的人肺上皮Beas-2B细胞炎性反应及氧化应激的作用。方法将人肺上皮Beas-2B细胞分为4组:对照组、LPS组、麦冬皂苷D+LPS组(简称OPD+LPS)和阳性对照组(简称DXMS+LPS)。CCK8法检测细胞增殖活性;ELISA检测肿瘤坏死因子α(tumor necrosis factorα,TNF-α)和白细胞介素6 (interleukin 6,IL-6)浓度;流式检测细胞活性氧(ROS)水平;比色法检测细胞内MDA浓度及髓过氧化物酶(MPO)、超氧化物歧化酶(SOD)和总抗氧化能力(T-AOC)活性;qRT-PCR检测细胞中IL-6、IL-10和IL-17 mRNA表达水平;Western blot检测细胞中核转录因子κB p65(NF-κB p65)及p-NF-κB p65蛋白表达水平。结果与对照组比较,LPS组Beas-2B细胞的SOD和T-AOC活性、IL-10 mRNA表达水平均显著降低(P<0.01),而MDA浓度、MPO活性、IL-6和IL-17 mRNA表达以及NF-κB p65和p-NF-κB p65蛋白表达均显著升高(P<0.01)。与LPS组比较,OPD+LPS组Beas-2B细胞的SOD和T-AOC活性、IL-10 mRNA表达水平均显著升高(P<0.05或P<0.01),而MDA浓度、MPO活性、IL-6和IL-17 mRNA表达以及NF-κB p65和p-NF-κB p65蛋白表达显著降低(P<0.01)。结论麦冬皂苷D预处理,可缓解LPS介导人肺上皮Beas-2B细胞的炎性反应与氧化应激。

Objective To investigate the effect of ophiopogonin D pretreatment on lipopolysaccharide(LPS)-mediated inflammatory response and oxidative stress in human pulmonary epithelial Beas-2 B cells.Methods Human pulmonary epithelial Beas-2 B cells were divided into four groups:control,LPS,ophiopogonin D+LPS(OPD+LPS),and positive control(DXMS+LPS).CCK8 assay was used to detect the cell proliferation activity.ELISA was used to detect the concentration levels of tumor necrosis factor α(TNF-α) and interleukin 6(IL-6).Flow cytometry was used to detect the reactive oxygen species(ROS) level in Beas-2 B cells.Colorimetry was used to detect the concentration of MDA and the activity of myeloperoxidase(MPO),superoxide dismutase(SOD) and total antioxide capacity(T-AOC).qRT-PCR was used to detect the mRNA expression of IL-6,IL-10 and IL-17.Western blot was used to detect the protein expression of nuclear factor κB p65(NF-κB p65) and p-NF-κB p65.Results Compared with the control group,SOD and T-AOC activities,IL-10 mRNA expression level of Beas-2 B cells in the LPS group were significantly decreased(P<0.01),while MDA concentration,MPO activity,IL-6 and IL-17 mRNA expression,and NF-κB p65 and p-NF-κB p65 protein expression were significantly increased(P<0.01).Compared with the LPS group,SOD and T-AOC activities,IL-10 mRNA expression level of Beas-2 B cells in the OPD+LPS group were significantly higher(P<0.05 or P<0.01),while MDA concentration,MPO activity,IL-6 and IL-17 mRNA expression,NF-κB p65 and p-NF-κB p65 protein expression were significantly lower(P<0.01).Conclusion Pretreatment with ophiopogonin D can alleviate the inflammatory response and oxidative stress of LPS mediated human pulmonary epithelial Beas-2 B cell.