Abstract
As chemical analysis for quality control (QC) of traditional Chinese medicine (TCM) formula is difficult to guarantee the effectiveness, a bioassay method that combines QC with evaluation of therapeutic effects has been developed to assess the TCM quality. Here, we chose a thirteen-component TCM formula, Lianhua Qingwen capsule (LHQW), as a representative sample, to explore the pivotal biomarkers for a bioassay and to investigate close association between QC and pharmacological actions. Initially, our results showed that chemical fingerprinting could not effectively distinguish batches of LHQW. Pharmacological experiments indicated that LHQW could treat influenza A virus (H1N1) infection in the H1N1 mouse model, as claimed in clinical trials, by improving pathologic alterations and bodyweight loss, and decreasing virus replication, lung lesions and inflammation. Furthermore, by using serum metabolomics analysis, we identified two important metabolites, prostaglandin F2α and arachidonic acid, and their metabolic pathway, arachidonic acid metabolism, as vital indicators of LHQW in treatment of influenza. Subsequently, macrophages transcriptomics highlighted the prominent role of cyclooxygenase-2 (COX-2) as the major rate-limiting enzyme in the arachidonic acid metabolism pathway. Finally, COX-2 was validated by in vivo gene expression and in vitro enzymatic activity with 43 batches of LHQW as a viable pharmacological biomarker for the establishment of bioassay-based QC. Our study provides systematic methodology in the pharmacological biomarker exploration for establishing the bioassay-based QC of LHQW or other TCM formulas relating to their pharmacological activities and mechanism.
Highlights
Traditional Chinese medicine (TCM) formula, as the primary form of TCM in the clinic, plays a vital role in the modernization of TCM (Lin et al, 2018)
MO, USA); interferon-gamma (IFN-g) was purchased from PeproTech (Rocky Hill, USA); high glucose Dulbecco's Modified Eagle's medium (DMEM) was purchased from HyClone (Logan, Utah, USA); fetal bovine serum and 0.25% trypsin were purchased from Gibco (Grand Island, New York, USA); standards of chlorogenic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid C, phillyrin, forsythiaside A and rutin were purchased from the Chengdu Pufei De Biotech., Ltd (Chengdu, China) and the purity of all these compounds was higher than 98.0%; High Performance Liquid Chromatography (HPLC)-grade methanol and acetonitrile were purchased from Merck (Darmstadt, Germany)
Lianhua Qingwen Capsule (LHQW) exhibits multiple pharmacological effects demonstrated in modern research including antiviral, antiinflammatory, immunomodulatory, and antipyretic actions, which have distinct therapeutic effects on diseases like influenza and pneumonia (Duan et al, 2011; Dong et al, 2014; Zhao et al, 2014; Ding et al, 2017)
Summary
Traditional Chinese medicine (TCM) formula, as the primary form of TCM in the clinic, plays a vital role in the modernization of TCM (Lin et al, 2018). The Botanical Drug Development Guidance for Industry issued by FDA in 2016 states that “because of the heterogeneous nature of a botanical drug and possible uncertainty about its active constituent, the technical challenges for quality control are to determine a botanical drug's identity and ensure its consistency of strength. It may require additional measures such as biological assays on the effect of variations, and a biological assay that reflects the drug's known or intended mechanism of action is preferred” (Food and Drug Administration, 2016). It is of value to explore the pivotal biomarkers for bioassays, and thereby, engage a close association between QC and pharmacological activity that has the potential for ensuring the consistency and clinical efficacy of TCM formulas
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