Mechanism of Jianpi Huatan Tongfu Prescription in Attenuating Mucus Hypersecretion in COPD Rats by Regulating Th17/Treg Balance

OBJECTIVE To investigate the mechanism by which the Jianpi Huatan Tongfu prescription (JPHT) regulates the Th17/Treg balance to suppress mucus hypersecretion in a rat model of chronic obstructive pulmonary disease(COPD).

METHODS This study first employed network pharmacology to predict potential targets and pathways of JPHT in treating mucus hypersecretion in COPD rats, followed by experimental validation. Forty SPF-grade male Wistar rats aged 6–8 weeks were randomly divided into 4 groups(n=10): blank control group(Ctrl group), COPD group, JPHT group, and salbutamol group(ALB group). A COPD rat model was established by intratracheal instillation of lipopolysaccharide combined with cigarette smoke exposure for 4 weeks. Drug interventions were administered starting the day after modeling and continued for 2 weeks. General conditions of the rats were observed, lung function was measured, and hematoxylin-eosin(HE) staining was used to examine morphological changes in lung tissue. Immunofluorescence was performed to detect the expression of MUC5AC, AQP5, and IL-17RA in lung tissues. Flow cytometry was used to analyze Th17/Treg ratios in spleen tissues. Western blotting was employed to measure the expression of NF-κB signaling pathway-related proteins and AQP5 in lung tissues.

RESULTS  Network pharmacology analysis indicated that the therapeutic effect of JPHT on COPD was associated with the IL-17A pathway. Compared with the Ctrl group, the COPD group exhibited poorer general conditions, decreased lung function, inflammatory cell infiltration, and emphysematous changes in the lung tissue. Flow cytometry revealed an imbalance in Th17/Treg ratio in the COPD group. Immunofluorescence showed upregulated expression of IL-17A and MUC5AC and downregulated expression of AQP5 in the COPD group. Western blotting demonstrated increased expression of NF-κB p65 and IKKβ and decreased expression of IκBα and AQP5 in the COPD group. Compared with the COPD group, JPHT treatment improved general conditions, lung function, and pathological damage in COPD rats, restored the Th17/Treg balance, suppressed the expression of IL-17A and NF-κB pathway-related proteins, upregulated AQP5 expression, downregulated MUC5AC expression, and alleviated pulmonary mucus hypersecretion.

CONCLUSION JPHT can restore the Th17/Treg balance, inhibit abnormal activation of the IL-17A/NF-κB pathway, regulate the expression of MUC5AC and AQP5, reduce pulmonary mucus secretion, and exert therapeutic effects on COPD.