Abstract
BackgroundHomeostasis is a crucial concept used to describe the condition of patients and the roles of herbs in traditional Chinese medicine. Qi-deficiency pattern is one of the conditions when loss of homeostasis and is usually characterized by symptoms including lassitude, spontaneous sweating, and a weak pulse, which are not easy to quantitate. Codonopsis pilosula and Astragalus membranaceus were usually prescribed for carriers with hepatitis and patients with metastatic colon cancer, because these patients tended to experience fatigue. However, crude drugs were prescribed based on the exterior symptoms of patients without controlling clinical setting, such as gender, age, and dietary habits. Limited molecular evidence of using gene expression as the guide for description is available. Therefore, the purpose of this study was to identify potential and objective biomarkers of these two qi-related drugs in a simplified cellular system.MethodsAqueous extracts of crude qi-tonifying herbs, C. pilosula and A. membranaceus, and that of a qi-consuming drug, Citrus reticulata, were prepared. Human liver cancer HepG2 cells were treated with the extracts of qi-tonifying herbs for 24 h. Differentially expressed genes were identified using microarrays and quantitative RT-PCR (qRT-PCR) and validated in two other hepatocellular cell lines, Huh7 and L-02.ResultsA total of 67 differentially expressed probes that responded to both herbs were identified. A pathway analysis revealed that these genes were involved in the development, growth, movement, and viability of the liver cells.ConclusionsAfter qRT-PCR validation and examination of clinical data from public domains, our results showed that two genes, GDF15 and HMOX1, could serve as biomarkers in liver cells for identifying responses after treatment with C. pilosula and A. membranaceus.
Highlights
Homeostasis is a crucial concept used to describe the condition of patients and the roles of herbs in traditional Chinese medicine
Quality control of three crude drugs The workflow for identifying differentially expressed genes in HepG2 cells treated with C. pilosula or A
By using the genomic approach of microarrays in hepatocellular carcinoma cells treated with qi-tonifying botanic drugs, C. pilosula and A. membranaceus, followed by quantitative RT-PCR (qRT-PCR) validation of hepatic normal cells and clinical samples, we identified drug-related genes associated with qi and their functions
Summary
Homeostasis is a crucial concept used to describe the condition of patients and the roles of herbs in traditional Chinese medicine. Qi-deficiency pattern is one of the conditions when loss of homeostasis and is usu‐ ally characterized by symptoms including lassitude, spontaneous sweating, and a weak pulse, which are not easy to quantitate. Aqueous extract decoction is a widely used form of drug preparation in traditional Chinese medicine (TCM), and physical symptoms in patients are a crucial basis for prescribing herbal decoctions. Qi-deficiency symptoms include shortness of breath, lassitude, listlessness, spontaneous sweating, a pale tongue, and a weak pulse [1]. The use of dried roots of Codonopsis pilosula and Astragalus membranaceus, qi-tonifying crude drugs, has been well documented in the ancient Chinese literatures, including in Ben cao gang mu [4], a few 100 years ago. An animal study demonstrated that C. pilosula polysaccharide attenuated sepsis in a mouse
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