Luan Xuejing, Zhu Dandan, Hao Yifei, Xie Jinghui, Wang Xiu, Li Yan, Zhu Jie
College of Integrated Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China.
Research Center of Integrated Traditional Chinese and Western Medicine, Wannan Medical College, Wuhu 241002, China.
Int Immunopharmacol. 2025 Jan 10;144:113636. doi: 10.1016/j.intimp.2024.113636. Epub 2024 Nov 22.
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.
The aim of this study is to investigate the modulatory effects of QBPF capsules on BMAL1 and the HIF-1α/glycolytic pathway in COPD.
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.
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.
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.
慢性阻塞性肺疾病（COPD）以持续炎症为特征。脑与肌肉芳香烃受体核转位蛋白样蛋白1（BMAL1）作为一种关键的生物钟基因，与糖酵解中上游因子缺氧诱导因子（HIF）-1α的表达水平相关，这可能影响COPD中炎症反应的发生。然而，启白平肺胶囊（QBPF）对BMAL1和HIF-1α/糖酵解途径的调节作用仍不清楚。
本研究旨在探讨QBPF胶囊对COPD中BMAL1以及HIF-1α/糖酵解途径的调节作用。
采用多因素方法构建COPD大鼠模型，包括强迫游泳、缺氧和四周香烟烟雾吸入。接下来，大鼠接受为期两周的药物灌胃治疗。最后，收集组织样本，使用各种分子生物学技术进行综合分析。这些方法包括分子对接、免疫沉淀、小干扰RNA（siRNA）、苏木精-伊红（HE）染色、蛋白质免疫印迹（WB）和免疫荧光等，以阐明QBPF在体外和体内治疗COPD的调节作用。
COPD中BMAL1的mRNA和蛋白表达水平降低，而HIF-1α的mRNA和蛋白含量增加。同时，己糖激酶（HK）、磷酸果糖激酶（PFK）、丙酮酸激酶（PK）、乳酸脱氢酶（LDH）和乳酸（LD）等糖酵解代谢产物的浓度升高，而ATP降低。QBPF胶囊可逆转BMAL1与HIF-1α之间的失衡，改善糖酵解途径紊乱，并减轻炎症反应。值得注意的是，在体内实验中，通过分子对接和免疫沉淀证实了BMAL1与HIF-1α之间的相互作用。在挽救实验中，对16HBE细胞用siRNA BMAL1干预后，BMAL1水平显著降低，QBPF的治疗效果也受到影响。
QBPF可上调生物钟基因BMAL1的表达水平，从而调节HIF-1α/糖酵解途径及代谢产物，改善COPD中的炎症反应。
