麦冬皂苷D对脂多糖诱导的肺泡上皮细胞凋亡的影响

Effect of Ophiopogonin D on lipopolysaccharide-induced apoptosis of alveolar epithelial cells

目的 探讨麦冬皂苷D调节白细胞介素-6(IL-6)/Janus激酶2(JAK2)/转录激活因子3(STAT3)信号通路对脂多糖(LPS)诱导的肺泡上皮细胞凋亡的影响。方法 将体外培养的A549 ATⅡ细胞均随机分为4组:对照组、LPS组、LPS+麦冬皂苷D组、LPS+麦冬皂苷D+colivelin(JAK2/STAT3信号激活药)组,除对照组外其余各组细胞建立损伤模型的同时以麦冬皂苷D和colivelin分组处理。以细胞计数试剂盒-8(CCK-8)实验、流式细胞术分别检测各组细胞增殖与凋亡情况;以蛋白质印迹法检测各组细胞IL-6/JAK2/STAT3信号通路蛋白表达水平。结果 对照组、LPS组、LPS+麦冬皂苷D组、LPS+麦冬皂苷D+colivelin组的A549细胞凋亡率分别为(2.52±0.73)%、(52.43±4.14)%、(1.67±0.52)%和(47.94±3.43)%,IL-6蛋白水平分别为0.14±0.03、0.49±0.05、0.17±0.04和0.45±0.06,p-JAK2/JAK2蛋白水平分别为0.17±0.04、0.64±0.08、0.19±0.06和0.61±0.07,p-STAT3/STAT3蛋白水平分别为0.20±0.06、0.69±0.10、0.22±0.07和0.65±0.09;ATⅡ细胞凋亡率分别为(3.01±0.69)%、(55.16±3.94)%、(2.35±0.71)%和(50.28±3.78)%,IL-6蛋白水平分别为0.11±0.03、0.87±0.13、0.19±0.04和0.84±0.12,p-JAK2/JAK2蛋白水平分别为0.13±0.04、0.56±0.08、0.15±0.03和0.53±0.07,p-STAT3/STAT3蛋白水平分别为0.30±0.08、0.79±0.14、0.33±0.09和0.75±0.13。上述指标:对照组、LPS+麦冬皂苷D组与LPS组比较,LPS+麦冬皂苷D+colivelin组与LPS+麦冬皂苷D组比较,在统计学上差异均有统计学意义(均P<0.05)。结论 麦冬皂苷D可通过抑制IL-6/JAK2/STAT3信号通路激活而降低LPS诱导的炎症与氧化应激水平,最终减轻LPS诱导的肺泡上皮细胞凋亡。

Objective To investigate the effect of Ophiopogonin D on lipopolysaccharide(LPS)-induced apoptosis of alveolar epithelial cells by regulating the interleukin-6(IL-6)/Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3) signaling pathway.Methods A549 AT Ⅱ cells cultured in vitro were randomly divided into four groups:control group,LPS group,LPS+Ophiopogonin D group,LPS+Ophiopogonin D+colivelin(JAK2/STAT3 signal activator)group,except for the control group,and cells in all other groups were established injury models while being grouped with Ophiopogonin D and colivelin for treatment.Cell counting kit-8(CCK-8) experiment and flow cytometry were applied to detect cell proliferation and apoptosis in each group;Western blotting was applied to detect the expression of IL-6/JAK2/STAT3signaling pathway proteins of cells in each group.Results The apoptosis rates of A549 cells in control group,LPS group,LPS+Ophiopogonin D group and LPS+Ophiopogonin D+colivelin group were(2.52±0.73)%,(52.43±4.14) %,(1.67±0.52) % and(47.94±3.43) %;IL-6 protein levels were 0.14±0.03,0.49±0.05,0.17±0.04 and 0.45±0.06,and p-JAK2/JAK2 protein levels were 0.17±0.04,0.64±0.08,0.19±0.06 and0.61±0.07;p-STAT3/STAT3 protein levels were 0.20±0.06,0.69±0.10,0.22±0.07 and 0.65±0.09;the apoptosis rates of AT Ⅱ cells were(3.01±0.69) %,(55.16±3.94) %,(2.35±0.71) % and(50.28±3.78) %;the levels of IL-6 protein were 0.11±0.03,0.87±0.13,0.19±0.04 and 0.84±0.12;the p-JAK2/JAK2protein levels were 0.13±0.04,0.56±0.08,0.15±0.03 and 0.53±0.07;p-STAT3/STAT3 protein levels were0.30±0.08,0.79±0.14,0.33±0.09 and 0.75±0.13.The above indexes:control group,LPS+Ophiopogonin D group compared with LPS group,LPS+Ophiopogonin D+colivelin group compared with LPS+Ophiopogonin D group,the differences were statistically significant(all P <0.05).Conclusion Ophiopogonin D can reduce LPS induced inflammation and oxidative stress levels by inhibiting the activation of IL-6/JAK2/STAT3 signaling pathway,ultimately reducing LPS-induced apoptosis of alveolar epithelial cells.