Kwan Kenneth K L, Huang Yun, Leung Ka W, Dong Tina T X, Tsim Karl W K
Shenzhen Key Laboratory of Edible and Medicinal Bioresources, Shenzhen Research Institute, Shenzhen, China.
Division of Life Science and Center for Chinese Medicine, The Hong Kong University of Science and Technology, Hong Kong, Hong Kong.
Front Pharmacol. 2019 Jun 21;10:614. doi: 10.3389/fphar.2019.00614. eCollection 2019.
Danggui Buxue Tang (DBT) is an ancient herbal mixture containing Astragali Radix and Angelicae Sinensis Radix, and which are commonly consumed for "qi-invigorating" (i.e., stimulating vital energy/energy metabolism) as traditional Chinese medicine (TCM). The pharmacological activities of DBT in anti-oxidation, estrogenic, hematopoietic, and immunogenic have been reported; however, the role of DBT in cellular energy metabolism has not been determined. Here, we employed an extracellular flux analyzer to evaluate the mitochondrial respiration of cultured H9C2 cardiomyoblasts in present of DBT. The herbal extract of DBT was qualified chemically for the major ingredients, i.e. astragaloside, calycosin, formononetin, Z-ligustilide, and ferulic acid. The anti-oxidant activities of DBT, as well as its major ingredients, were determined by Folin-Ciocalteu assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, and protective effect in tert-butyl hydroperoxide (tBHP)-treated cultured cardiomyoblasts. In addition, a real-time oxygen consumption rate (OCR) in herbal extract-treated cultured cardiomyoblasts was revealed by using a Seahorse extracellular flux analyzer. In addition, the transcript expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PCG-1α) and other genes relating to mitochondria biogenesis were determined in cardiomyoblasts under different herbal treatments. DBT possessed the strongest anti-oxidant activity and protective effects on the oxidatively stressed cardiomyoblasts. By revealing the OCR in mitochondria, the health state of cultured cardiomyoblasts under DBT was improved increase of basal respiration, proton leak, non-mitochondria, and adenosine triphosphate (ATP) production. Furthermore, the transcriptional activities of genes responsible for mitochondrial biogenesis and DNA replication were stimulated by application of DBT in cultures.
当归补血汤（DBT）是一种古老的草药合剂，包含黄芪和当归，作为传统中药常用于“补气”（即刺激元气/能量代谢）。DBT在抗氧化、雌激素样、造血和免疫原性方面的药理活性已有报道；然而，DBT在细胞能量代谢中的作用尚未确定。在此，我们使用细胞外通量分析仪评估DBT存在时培养的H9C2心肌成纤维细胞的线粒体呼吸。对DBT的草药提取物进行了主要成分的化学鉴定，即黄芪甲苷、毛蕊异黄酮、芒柄花素、Z-藁本内酯和阿魏酸。通过福林-酚试剂法、2,2-二苯基-1-苦基肼（DPPH）自由基清除试验以及在叔丁基过氧化氢（tBHP）处理的培养心肌成纤维细胞中的保护作用，测定了DBT及其主要成分的抗氧化活性。此外，使用海马细胞外通量分析仪揭示了草药提取物处理的培养心肌成纤维细胞中的实时耗氧率（OCR）。此外，在不同草药处理下的心肌成纤维细胞中，测定了过氧化物酶体增殖物激活受体γ共激活因子1α（PCG-1α）和其他与线粒体生物发生相关基因的转录表达。DBT对氧化应激的心肌成纤维细胞具有最强的抗氧化活性和保护作用。通过揭示线粒体中的OCR，DBT作用下培养的心肌成纤维细胞的健康状态得到改善，基础呼吸、质子泄漏、非线粒体呼吸和三磷酸腺苷（ATP）产生增加。此外，在培养物中应用DBT可刺激负责线粒体生物发生和DNA复制的基因的转录活性。
