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.
ABSTRACT
Objective: To explore the effect of Qingjin Huatan Tang (QJHTD) on the inflammatory response of chronic obstructive pulmonary disease(COPD) rats by observing the autophagy regulating effect of QJHTD on COPD rats. Method: The 50 SPF grade male rats were randomly divided into 5 groups, with 10 rats in each group. In addition to the normal group, the remaining 40 male rats were randomly divided into 5 groups. After the establishment of the hematoxylin and eosin(HE) staining identification model, the drugs were given to the 5 groups by gavage for 2 weeks, high and low-dose QJHTD groups were give the drug at 30, 10 g·kg-1. Roxithromycin positive control group was given the drug at 0.017 5 g·kg-1. The model control group and the normal group were given the same volume of normal saline. At 1 h after the last gavage, the rats were put to death to extract the airway, and the expressions of autophagy microtuble-associated protein light chain 3 (LC3),Beclin-1 were detected by Real-time quantitative PCR (Real-time PCR) and Western blot. Changes of inflammatory cytokines interleukin-6 (IL-6) and interleukin-8(IL-8) were detected by enzyme linked immunosorbent assay (ELISA). Result: Real-time PCR analysis showed that compared with the normal group, Beclin-1 and LC3 mRNA expressions of autophagy factors in the model group were increased to varying degrees(PPPPPConclusion: QJHTD can alleviate the bronchial inflammation in COPD rats, and its mechanism may be related to the inhibition of autophagy in airway epithelium by QJHTD.