Qibai Pingfei Capsule ameliorated inflammation in chronic obstructive pulmonary disease (COPD) via HIF-1 α/glycolysis pathway mediated of BMAL1.

BACKGROUND

Chronic obstructive pulmonary disease (COPD) is characterized for the persistent inflammation. The brain and muscle arnt-like 1 (BMAL1), as a crucial clock gene, is associated with the expression level of upstream factor hypoxia-inducible factor (HIF)-1α in glycolysis, which may affect the occurrence of inflammatory reactions in COPD. However, the moderation effect of Qibai Pingfei Capsule (QBPF) capsule is still unknown on BMAL1 and HIF-1α/glycolytic pathway.

OBJECTIVE

The aim of this study is to investigate the modulatory effects of QBPF capsules on BMAL1 and the HIF-1α/glycolytic pathway in COPD.

METHODS

The multifactorial approach were used to construct the COPD rat model, including forced swimming, hypoxia, and inhalation of cigarette smoke with four weeks. Nextly, the rats received a two-week course of gavage treatment with medicant. Finally, tissue samples were collected for comprehensive analysis using various molecular biology techniques. These methods included molecular docking, immunoprecipitation, small interfering RNA (siRNA), hematoxylin and eosin (HE) staining, western blot (WB), and immunofluorescence etc. to elucidate the modulatory effects of QBPF for treating COPD in vitro and in vivo.

RESULTS

The expression levels in mRNA and protein of BMAL1 decreased in COPD, while the content in mRNA and protein of HIF-1α increased. At the same time, the concentration in glycolytic metabolites of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), lactate dehydrogenase (LDH), and lactate (LD) increased, and ATP decreased. The QBPF capsule can reverse the imbalance between BMAL1 and HIF-1α, improve disorders of glycolytic pathway, and alleviate the inflammation response. Notably, in vivo experiments, the interaction between BMAL1 and HIF-1α were confirmed via molecular docking and immunoprecipitation. In rescue experiments, intervention with siRNA BMAL1 in 16HBE cells revealed a significant decrease in BMAL1 levels and the therapeutic effect of QBPF was also affected.

CONCLUSION

QBPF could up-regulate the expression level of clock gene BMAL1, thereby regulating the HIF-1α/glycolytic pathway and metabolite to improve the inflammatory response in COPD.