Danggui Buxue Tang, a Chinese Herbal Decoction Containing Astragali Radix and Angelicae Sinensis Radix, Modulates Mitochondrial Bioenergetics in Cultured Cardiomyoblasts.

Kenneth K.L Kwan,Ka W Leung,Yun Huang,Tina T.X Dong,Karl W.K Tsim

doi:10.3389/fphar.2019.00614

Abstract

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 via 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.

Highlights

Traditional Chinese medicine (TCM) has been used as medicine, or health supplement, in China over thousands of years (Lu and Lu, 2014)
In chemical standardization of herbal extracts, an optimized LC-MS method was used to determine the amounts of major ingredients within Danggui Buxue Tang (DBT), e.g. astragaloside IV, calycosin, formononetin, ferulic acid, and Z-ligustilide, as reported (Zheng et al, 2012; Supplementary Figure 1A)
We have previously reported the protective effects of DBT in cardiomyocyte’s energy metabolism in mice having acute myocardial infarction or cardiac ischemia-reperfusion injury: this protective effect was triggered by mitochondrial enhancement as well as glutathione status in red blood cell (Mak et al, 2006)

Summary

Introduction

Traditional Chinese medicine (TCM) has been used as medicine, or health supplement, in China over thousands of years (Lu and Lu, 2014). TCM herbal decoction is prepared by a unique methodology with specific combination of different herbs as a formula. Among thousands of herbal formulae, Danggui Buxue Tang (DBT) is an ancient Chinese herbal decoction having a simple combination of two herbs. DBT contains two common herbs, Astragali Radix [AR; roots of Astragalus membranaceus (Fisch). Mongholicus (Bunge) Hsiao] and Angelicae Sinensis Radix (ASR; roots of Angelica sinensis Oliv). DBT is a well-known herbal decoction in clinical treatment for women suffering from menopausal symptoms (Gong et al, 2016a). According to TCM theory, DBT promotes “qi” (vital energy) enhancement and “blood” (body circulation) nourishment. DBT has been proposed to possess pharmacological properties both in vivo and in vitro, which includes: i) stimulating hematopoietic function (Yim et al, 2000), ii) exhibiting estrogenic property (Choi et al, 2011), iii) accelerating bone regeneration (Wang et al, 2015), iv) promoting immune system (Yang et al, 2014), v) protecting cardiovascular and pulmonary system (Mak et al, 2006), and vi) engaging formation of capillary and blood vessel (Gong et al, 2016a)

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Conclusion