Effect of Qingjin Huatantang on AECOPD Inflammatory Factors by Regulating p62 / 中国实验方剂学杂志

ABSTRACT

Objective:To regulate autophagy protein p62 of airway epithelial cells in acute exacerbation of chronic obstructive pulmonary disease (AECOPD) rats with Qingjin Huatantang, in order to explore its effect on interleukin (IL) -1β and tumor necrosis, tumor necrosis factor-α (TNF-α), leukotriene B4 (LTB4), and leukotriene C4 (LTC4). Method:Airway epithelial cells and AECOPD airway epithelial cells were cultured. Sixty SPF male SD rats were randomly divided into 6 groups, namely high, medium and low-dose Qingjin Huatantang groups, western medicine group, model group and normal group. Except for the normal group, the remaining groups were included into the AECOPD model by lipopolysaccharide (LPS) tracheal instillation method + fumigation method. After modeling, the dosage of the high-dose traditional Chinese medicine group was 30 g·kg-1·d-1, that of the middle-dose group was 15 g·kg-1·d-1, that of the low-dose group was 7.5 g·kg-1·d-1, the positive control group was given luo erythromycin (0.017 5 g·kg-1·d-1), the model group and the blank control group were orally given normal saline with the volume of 20 mL·kg-1·d-1. Serum was extracted two weeks after administration, and the cells were intervened with drug-containing serum. The content of interleukin IL-1β, TNF-α, and LTB4 in cell supernatants were detected by enzyme-linked immunosorbent assay (ELISA). And LTC4 content, p62 mRNA and protein expressions in lung airway epithelial cells were detected by quantitative real-time fluorescence polymerase chain reaction (Real-time PCR) and Western blot. Result:Compared with the normal group, ELISA results showed that IL-1β, TNF-α, LTB4, and LTC4 in the model group were significantly increased (P<0.01). Compared with the model group, IL-1β, TNF-α, LTB4, LTC4 in cell supernatants in each administration group were significantly reduced (P<0.01), mRNA and protein expressions in p62 showed that compared with the normal group, mRNA and protein expressions in p62 of model group significantly decreased (P<0.01). Compared with the model group, the mRNA and protein expressions of p62 in each administration group significantly increased to different degrees (P<0.01). The expression of autophagy in Qingjin Huatantang high-dose group and western medicine group was comparable. Conclusion:Qingjin Huatantang can reduce the inflammatory response in airway epithelial cells, which may be related to the regulation of autophagy protein p62.