五味子乙素对香烟烟雾提取物和脂多糖损伤的肺上皮细胞干预作用研究

Effect of Schisandrin B on cigarette smoke extract and lipopolysaccharide injury in lung epithelial cells

目的:观察五味子乙素(Sch B)对慢性阻塞性肺疾病(COPD)急性加重期细胞模型的影响,并初步探讨其作用机制。方法:用香烟烟雾提取物(CSE)联合脂多糖(LPS)作用于肺上皮细胞株A549,制备COPD急性加重期体外模型,并给予相应浓度的五味子乙素进行干预,通过MTT法测定各组细胞存活率、RT-PCR法测定细胞炎症因子白细胞介素8(IL-8)、环氧合酶2(COX-2)的mRNA水平; ELISA法测定细胞上清中IL-8、COX-2水平; Western blot法测定细胞磷酸化NF-κB蛋白水平。结果:4%CSE联合0. 1μg/ml LPS作用24 h能够促进肺上皮细胞增殖,炎症因子表达明显上升,给予Sch B能够明显抑制CSE联合LPS对细胞的增殖作用,改善细胞炎症状态;降低损伤细胞IL-8、COX-2 mRNA表达水平;减少细胞上清中IL-8、COX-2含量;降低细胞中磷酸化NF-κB的蛋白表达水平。结论:Sch B可改善CSE联合LPS引起的肺上皮细胞炎症,其作用机制可能与抑制肺上皮细胞增殖,抑制炎症因子表达,发挥抗炎作用有关。该研究为Sch B应用于COPD急性加重期的临床治疗提供实验依据。

Objective:To explore the effect of Schisandrin B(Sch B)on cigarette smoke extract(CSE)combined with lipopolysaccharide(LPS)injury in lung epithelial cells of chronic obstructive pulmonary disease(COPD)and to explore its mechanism.Methods:By using CSE combined with LPS induced injury of pulmonary epithelial cell line A549 as acute exacerbation model of COPD in vitro;and proper concentration of Sch B was given to prevent the injury of CSE and LPS,then the cell viability was determined by MTT assay,mRNA levels of interleukin-8(IL-8)and cyclooxygenase-2(COX-2)were measured by RT-PCR,the levels of IL-8 and COX-2 in supernatant were measured by ELISA,the level of phosphorylated NF-κB protein was measured by Western blot.Results:Co-cultured with 4%CSE combined with 0.1μg/ml LPS for 24 h,lung epithelial cell proliferation was promoted,inflammatory factor significantly increased,Sch B could significantly inhibit the proliferation of lung epithelial cells and improve the inflammation induced by CSE combined with LPS,reduce the expression of IL-8 and COX-2 mRNA in injured cells,decrease IL-8 and COX-2 levels in the supernatant,reduce the level of phosphorylated NF-κB protein expression in cells.Conclusion:Sch B can inhibit the proliferation of lung epithelial cells,and the inflammatory factor expression induced by CSE combined with LPS,which provides experimental evidence for the clinical application of Sch B in the treatment of acute exacerbation of COPD.