北京中医药大学学报2017，Vol.40Issue(4)：314-321,8.DOI:10.3969/j.issn.1006-2157.2017.04.010
益肺活血方对香烟提取物诱导巨噬细胞炎症反应的影响
Effects of Yifei Huoxue Fang on the inflammatory response in macrophages induced by cigarette smoke extract
摘要
Abstract
Objective To explore the effects of Yifei Huoxue Fang (YFHXF, lung-invigorating blood-activating formula) on the inflammatory response in macrophages induced by cigarette smoke extract.Methods U937 monocytic cells were differentiated into macrophages using phorbol 12-myristate 13-acetate (PMA).These PMA-induced macrophages were then treated with YFHXF at various concentrations (1.2,2.4,4.8,9.6 g/L) or exposed to cigarette smoke extract (CSE)(5,10,20,50,100 mL/L).At 12, 24, and 48 h, viability of the cells was assessed by using the cell counting kit-8 (CCK-8).Macrophages were divided into 5 groups: normal group, CSE group, CSE+ YFHXF low-dose (1.2 g/L) group, CSE+ YFHXF medium-dose (2.4 g/L) group, and CSE+ YFHXF high-dose (4.8 g/L) group.Levels of IL-8 and TNF-α in cell culture supernatant were detected by using enzyme-linked immunosorbent assay (ELISA), and gene expression of IL-8 and TNF-α were also measured with real-time polymerase chain reaction (RT-PCR).For the study of mechanism, macrophages were divided into 4 groups: normal group, CSE group, CSE+ YFHXF (2.4 g/L) group, and CSE+NF-κB inhibifant(parthenolide) group.The expression of nuclear factor-κB (NF-κB) p50/p65 and p-IκB was examined by using Western blot method.Meanwhile, the expression of nucleus p65 and plasma p-IκB in macrophages was also measured by using the immunofluorescent assay.Results Cytotoxicity was present in macrophages at all timing points induced by CSE (100 ml/L) and at 24, and 48 hours after treated with YFHXF (9.6 g/L).Compared with normal cell group, IL-8 and TNF-α of CSE group, and NF-κB p50/p65 and p-IκB expression were all upregulated (P<0.05).NF-κB nuclear translocation was increased (P<0.05).Yifei Huoxue Fang could inhibit the expression of IL-8 and TNF-α induced by CSE, and viability of NF-κB (P<0.05).Conclusion Yifei Huoxue Fang could inhibit the expression of IL-8 and TNF-α induced by CSE, possibly by inhibiting NF-κB activation.关键词
慢性阻塞性肺疾病/益肺活血方/巨噬细胞/白细胞介素-8/肿瘤坏死因子-α/核转录因子-κBKey words
chronic obstructive pulmonary disease/Yifei Huoxue Formula/macrophages/IL-8/TNF-α/nuclear factor-κB分类
医药卫生引用本文复制引用
简芙平,欧敏..益肺活血方对香烟提取物诱导巨噬细胞炎症反应的影响[J].北京中医药大学学报,2017,40(4):314-321,8.基金项目
军队中医药科研专项基金项目(No.10ZYZ219) （No.10ZYZ219）
