Ren Hui, Wu Wenxing, Chen Jiangyan, Li Quan, Wang Hengbin, Qian Dawei, Guo Sheng, Duan Jin-Ao
National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization and Jiangsu Key Laboratory for High Technology Research of Traditional Chinese Medicine Formulae, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
Leiyunshang Pharmaceutical Co. Limited, Suzhou, 215003, China.
J Ethnopharmacol. 2024 Apr 24;324:117816. doi: 10.1016/j.jep.2024.117816. Epub 2024 Jan 28.
Bufei Huoxue capsule (BHC) as a classic Chinese patent medicine formula, has the efficacy of tonifying the lungs and activating the blood. It has been extensively used in China for the treatment of chronic obstructive pulmonary disease (COPD) clinically. However, its mechanism is still unclear, which hampers the applications of BHC in treating COPD.
The purpose of the present study was to demonstrate the protective efficacy and mechanism of BHC on COPD model rats by integrating serum metabolomics analysis and network pharmacology study.
A COPD rat model was established by cigarette fumigation combined with lipopolysaccharide (LPS) airway drip for 90 consecutive days. After oral administration for 30 days, the rats were placed in the body tracing box of the EMKA Small Animal Noninvasive Lung Function Test System to determine lung function related indexes. Histopathological alteration was observed by H&E staining and Masson staining. The serum levels of inflammatory cytokine, matrix metalloprotein 9, and laminin were determined by ELISA kits. Oxidative stress levels were tested by biochemical methods. UHPLC-Q-TOF/MS analysis of serum metabolomics and network pharmacology were performed to reveal the bioactive metabolites, key components and pathways for BHC treating COPD. WB and ELISA kits were used to verify the effects of BHC on key pathway.
BHC could improve lung function, immunity, lung histopathological changes and collagen deposition in COPD model rats. It also could significantly reduce inflammatory response in vivo, regulate oxidative stress level, reduce laminin content, and regulate protease-antiprotease balance. Metabolomics analysis found 46 biomarkers of COPD, of which BHC significantly improved the levels of 23 differential metabolites including arachidonic acid, leukotriene B4 and prostaglandin E2. Combined with the results of network pharmacology, the components of BHC, such as calycosin, oxypaeoniflora, (S)-bavachin and neobavaisoflavone could play therapeutic roles through the arachidonic acid pathway. In addition, the results of WB and ELISA indicated that BHC could suppress the expressions of COX2 and 5-LOX in lung tissues and inhibit the generation of AA and its metabolites in serum samples. Regulation of arachidonic acid metabolic pathway may be the crucial mechanism for BHC treating COPD.
In summary, the studies indicated that BHC exhibited the protective effect on COPD model rats by anti-inflammatory and anti-oxidative properties through arachidonic acid metabolism pathway. This study provided beneficial support for the applications of BHC in treating COPD.
补肺活血胶囊（BHC）作为一种经典的中成药配方，具有补肺活血的功效。在中国，它已被广泛应用于慢性阻塞性肺疾病（COPD）的临床治疗。然而，其作用机制仍不清楚，这阻碍了BHC在治疗COPD中的应用。
本研究旨在通过整合血清代谢组学分析和网络药理学研究，阐明BHC对COPD模型大鼠的保护作用及其机制。
采用香烟熏吸联合脂多糖（LPS）气道滴注法连续90天建立COPD大鼠模型。给药30天后，将大鼠置于EMKA小动物无创肺功能测试系统的体描箱中测定肺功能相关指标。通过苏木精-伊红（H&E）染色和Masson染色观察组织病理学改变。采用酶联免疫吸附测定（ELISA）试剂盒测定血清炎症细胞因子、基质金属蛋白酶9和层粘连蛋白水平。采用生化方法检测氧化应激水平。进行血清代谢组学的超高效液相色谱-四极杆飞行时间质谱（UHPLC-Q-TOF/MS）分析和网络药理学研究，以揭示BHC治疗COPD的生物活性代谢产物、关键成分和途径。采用蛋白质免疫印迹法（WB）和ELISA试剂盒验证BHC对关键途径的作用。
BHC可改善COPD模型大鼠的肺功能、免疫力、肺组织病理学变化及胶原沉积。它还能显著减轻体内炎症反应，调节氧化应激水平，降低层粘连蛋白含量，调节蛋白酶-抗蛋白酶平衡。代谢组学分析发现了46种COPD生物标志物，其中BHC显著提高了23种差异代谢物的水平，包括花生四烯酸、白三烯B4和前列腺素E2。结合网络药理学结果，BHC的成分，如毛蕊异黄酮、芍药苷、（S）-补骨脂素和新补骨脂异黄酮可能通过花生四烯酸途径发挥治疗作用。此外，WB和ELISA结果表明，BHC可抑制肺组织中COX2和5-LOX的表达，并抑制血清样本中花生四烯酸及其代谢产物的生成。调节花生四烯酸代谢途径可能是BHC治疗COPD的关键机制。
综上所述，研究表明BHC通过花生四烯酸代谢途径的抗炎和抗氧化特性对COPD模型大鼠具有保护作用。本研究为BHC在治疗COPD中的应用提供了有益的支持。
