主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

JOURNAL OF BEIJIGN UNIVERSITY OF TRADITIONAL CHINE ›› 2017, Vol. 40 ›› Issue (4): 314-321.doi: 10.3969/j.issn.1006-2157.2017.04.010

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Effects of Yifei Huoxue Fang on the inflammatory response in macrophages induced by cigarette smoke extract*

JIAN Fuping1 OU Min1,2#   

  1. 1 The Third Clinical Medical School of Southern Medical University, Guangdong 510515;
    2 VIP Department of Respiration Disease, Navy General Hospital of PLA
  • Received:2016-11-04 Online:2017-04-30 Published:2017-04-30

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.

Key words: chronic obstructive pulmonary disease, Yifei Huoxue Formula, macrophages, IL-8, TNF-α, nuclear factor-κB

CLC Number: 

  • R285.5