Ginseng, the root and rhizome of Panax ginseng C. A. Mey. (Araliaceae), is one of the most famed dietary and medicinal herbs worldwide due to its multifaceted efficacies. Ginsenosides and carbohydrates are demonstrated the major bioactive components of ginseng. Ginseng materials are always formed under various conditions, e.g. different growth years or different post-harvest processing/handling manners. These conditions can impact chemical profiles and thereby cause different quality and efficacy of ginseng. To address this issue, it will be necessary to understand the correlation between the quality and efficacy of ginseng materials formed under different conditions. Previous studies have attempted to investigate how growth years and post-harvest processing/handling manners affect the quality and efficacy of ginseng. In the most of these cases, several chemical components and biological parameters were selected as the indicators for evaluating the quality and efficacy of ginseng, respectively. However, it has been well recognized that the therapy of ginseng is featured by "multiple components against multiple targets". Therefore, several selected indicators may fail to comprehensively characterize the quality and efficacy of ginseng, thus cannot accurately reveal their correlations. Instead, holism-based approaches should be employed. In this study, we integrated chemomics, metabolomics and gut microbiota genomics to investigate the correlation between the quality and efficacy of ginseng in the conditions of growth years, steam-processing and sulfur-fumigation. First, chemomics approach was developed to qualitatively and quantitatively determine major ginsenosides and carbohydrates (poly-, oligo- and monosaccharides) by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) and high performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) for characterizing the overall quality of ginseng. Second, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS)-based metabolomics and 16S rRNA gene sequencing-based gut microbiota genomics coupled with biochemical parameters determination were performed to evaluate anti-fatigue and anti-obesity activities of the different ginseng on animal models. Third, the obtained multi-omics data were processed by multivariate statistical analysis and then were integrated to discuss the correlation between the quality and efficacy of ginseng materials in different conditions. The results indicated that: 1) ginseng with 4-6 growth years possessed different anti-fatigue activity in multiple targets due to the different effects of ginsenosides and carbohydrates on endogenous metabolism and gut microbiota; 2) steam-processing qualitatively and quantitatively altered ginsenosides and carbohydrates in ginseng, resulting in different anti-obesity activity between white ginseng and red ginseng, and the mechanisms potentially involve chemically structural/compositional specificity to gut microbiota; 3) SO2 residual content caused by sulfur-fumigation did not correlate with the quality, efficacy and toxicity changes of sulfur-fumigated ginseng, more specifically, less SO2 residue did not indicate higher quality, better efficacy nor weaker toxicity. The research provides scientific insights for guiding the clinical and dietary practice of ginseng and offers new methodology for comprehensively exploring the correlation between the quality and efficacy of herbal medicines

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

published_or_final_version / Biological Sciences / Master / Master of Philosophy

Matrix metalloproteinase (MMP) induced extra cellular matrix (ECM) degradation is a crucial process involved in the development of many chronic inflammatory diseases, including cardiac remodeling and cancer metastasis. In cardiac remodeling, the presence of pathological stimuli leads to elevated MMP-9 expression and impairment of cardiac performance, which subsequently develops into heart failure. While in tumorgenesis, MMP-9 has been found to play key roles in metastasis, as it can break physical barriers for the tumor. Therefore, searching for agents targeting MMP-9 is a new direction for the treatment of cardiac remodeling and cancer metastasis. Chinese herbal medicine is becoming increasingly used worldwide in recent decades. In the past twenty years, as many highly selective and sensitive bioassays were introduced into the bioactive compounds screening from herbal medicine, more than one hundred new drug candidates have been identified. Therefore, herbal medicine is a potential source of bioactive compounds. Panax notoginseng (PNG) is one of the most common traditional Chinese medicines to treat cardiovascular diseases, and it was also reported to have anti-cancer effect. We hypothesized that it contains bioactive compounds that could inhibit MMP-9 activity in cardiomyocytes and cancer cells. In order to examine the effect of PNG on cardiac remodeling and cancer metastasis, we employed TNF-α induced MMP-9 in H9c2 cell (a rat cardiomyocyte) and HepG-2 cell (a human hepatoma cell) as an in vitro assay, respectively. PNG was first extracted by four different extraction methods according to the polarity of the solvent. The most effective fraction in suppressing MMP-9 activity in TNF-α induced H9c2 cell was chosen for further separation by silica gel column chromatography and high performance liquid chromatography (HPLC) until a single compound was isolated. According to the result of spectroscopic analysis by NMR, the compound was identified as ginsenoside Rb1. For the bioactivity assays, real-time quantitative polymerase chain reaction (QPCR) and Enzyme-linked immunosorbent assay (ELISA) were used to measure the mRNA and protein expression of MMP-9, respectively. We also examined the MMP-9 activity by gelatin zymography. The results showed that both of the PNG extract obtained from 10% ethanol extraction method (PNG-3) and purified Compound P (ginsenoside Rb1) showed significant inhibitory effect on MMP-9 expression and activity in H9c2 cells and HepG-2 cells. We further examined the molecular mechanisms of the inhibitory effect of PNG-3. H9c2 and HepG-2 cells were pretreated with different kinase inhibitors followed by the activation by TNF-α. The results showed the protein kinase R (PKR) inhibitor could inhibit TNF-α induced MMP-9 in both of the two cell lines. Furthermore, the results of Western blot showed the PNG-3 suppressed the phosphorylation of eIF-2α which is a down-stream effector of PKR in TNF-α stimulated H9c2 and HepG-2 cells, respectively. Therefore, PNG-3 may act through PKR to regulate TNF-α induced MMP-9 activity. In summary, bioactivity guided fractionation is an effective way of isolating bioactive compounds from medicinal herbs. In addition, PNG containing ginsenoside Rb1 may be a potential candidate of MMP-9 inhibition for the treatment of cardiac remodeling and cancer metastasis. / published_or_final_version / Paediatrics and Adolescent Medicine / Master / Master of Philosophy

abstract / Biological Sciences / Master / Master of Philosophy

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

published_or_final_version / Anatomy / Master / Master of Philosophy

University of Macau / Institute of Chinese Medical Sciences