A systematic metabolic pathway identification of Common Gardenia Fruit (Gardeniae Fructus) in mouse bile, plasma, urine and feces by HPLC-Q-TOF-MS/MS.

Gardeniae Fructus was a traditional Chinese medicine (TCM) containing various biological ingredients including iridoids and crocetins, monocyclic monoterpenes, organic acids, and flavonoids. However, few systematic identification studies of the bioactive components in vivo have been reported. Herein, the ingredients and metabolites of Gardeniae Fructus were investigated using high-performance liquid chromatography coupled with high-sensitivity Q-TOF mass spectrometry. A total of 45 prototype compounds in Gardeniae Fructus extract were tentatively identified. After oral administration, 69 of prototypes and metabolites were identified from mice bile, plasma, urine, and feces, in which, 31 compounds were prototypes, and 38 chemicals were metabolites. The in vivo biotransformation pathways of these metabolites were also proposed including phase I (hydrolysis, hydrogenation, oxidation, loss of O, and ketone formation, decarboxylation) and phase II reactions (glycine, cysteine, glutathione, and glutamine, and sulfate conjugation, and glucuronidation). For the first time, our results had revealed systematic metabolic profiles of ingredients in Gardeniae Fructus extract in vivo of mice and replenished novel knowledge into the explanation of effective material and/or toxicological basis of Gardeniae Fructus which deserves further investigation.

Green synthesis and characterization of biologically active nanosilver from seed extract of Gardenia jasminoides Ellis.

This article reports the utilization of seed extract (GSE) from Gardenia jasminoides Ellis. in the synthesis of silver nanoparticles (Gs-AgNPs) with versatile biological activities. The synthesized Gs-AgNPs were spherical in shape, crystal lattice with an average size of 20 nm as confirmed by UV-vis spectrum, X-ray diffractometer (XRD), Transmission electron microscopy with Energy dispersive X-ray spectroscopy (TEM-EDS) and particle size analyses (PSA). Phenolic compounds, proteins, and terpenoids were likely involved in the Gs-AgNPs synthesis, as indicated by Fourier-transform infrared spectroscopy (FTIR) analysis. The minimum bactericidal concentration (MBC) of the Gs-AgNPs was 12.5 µg·ml-1 for S. enterica Typhimurium and 10 µg·ml-1 for S. aureus. The MBC of the Gs-AgNPs induced >70% bacterial cell death within 60 min, as confirmed by growth curve analysis followed by Confocal laser scanning microscope (CLSM). Gs-AgNPs showed the highest scavenging activity for 1, 2-diphenyl-1-picrylhydrazyl DPPH radical (92.3 ±â€¯0.86%), Nitric oxide (NO) radical (72.5 ±â€¯2.15%), and Hydrogen peroxide H2O2 radical (85.25 ±â€¯1.45%). Anticancer results revealed an IC50 of 15.625 ±â€¯1.3 µg·ml-1 for Gs-AgNPs, whereas it was 580.54 ±â€¯2.5 µg·ml-1 for GSE. The Gs-AgNPs generated high reactive oxygen species (ROS) resulting in induced apoptosis as evident by up-regulation of apoptosis-related protein. In addition, the photocatalytic results revealed about 92% of the reduction in Coomassie Brilliant Blue dye color with Gs-AgNPs. Hence, this work provided economically viable and ecologically sustainable Gs-AgNPs as an alternative biomaterial for future therapeutic applications as antimicrobial, antioxidant, anti-cancer agents and in dye degradation for water remediation.

Antihyperlipidemic and hepatoprotective effects of Gardenin A in cellular and high fat diet fed rodent models.

The gum of Gardenia resinifera Roth., is one of the important drugs used in the Indian system of medicine and a source of unique polymethoxylated flavones. This study was aimed to evaluate the antihyperlipidemic and anti-NAFLD effects of Gardenin A (Gar-A) from G. resinifera gum using in vitro and in vivo models. Gar-A was isolated from G. resinifera gum and was identified on the basis of the physical and spectral data. Toxicity of Gar-A to HepG2 cells was evaluated using MTT assay. The ability of Gar-A to reduce steatosis was assessed using oleate-palmitate induced HepG2 cell lines by estimating the lipid levels by ORO staining and by estimating the intracellular triglyceride content. Effect of Gar-A on amelioration of lipotoxicity was measured by estimating the LDH levels. The doses for in vivo experiments were fixed by Irwin test, between 50 and 100 mg/kg concentrations, through oral route. The acute antihyperlipidemic effect of Gar-A was assessed in Triton WR-1339 induced hyperlipidemic animals. The chronic antihyperlipidemic and anti-NAFLD effects of Gar-A were evaluated in HFD fed rats. In vitro experiments with HepG2 cell line indicated that the cells treated with Gar-A did not show any significant reduction in the viability up to 70 µg/mL concentration. Steatotic HepG2 cells treated with Gar-A showed a significant reduction in lipid accumulation at 2.5-10 µg/mL concentrations. In triton induced hyperlipidemic rats, the treatment significantly reduced the lipid levels at the synthesis phase. The treatment with Gar-A to the HFD fed animals significantly lowered the steatosis and transaminase levels. The other biochemical parameters such as TC, TG, LDL-c, ALP and ACP were also decreased significantly. Treatment with Gar-A significantly lowered the hyperlipidemia and fat accumulation in the liver; detailed molecular investigations are necessary to establish the antihyperlipidemic and hepatoprotective potentials of Gar-A.

Ethanol extract of baked Gardeniae Fructus exhibits in vitro and in vivo anti-metastatic and anti-angiogenic activities in malignant cancer cells: Role of suppression of the NF-κB and HIF-1α pathways.

Gardeniae Fructus (GF, Zhi Zi in China), a fruit of Gardenia jasminoides Ellis, has been used in traditional medicine to reduce inflammation and headache and to treat hepatic disorders, hypertension, and icterus. In recent studies, extract of raw or stir-baked GF was shown to have pharmacological activities for viral infection, thrombosis, hyperlipidemia, convulsion, inflammation, oxidative stress, and others. In addition, baked GF extract suppressed the proteolytic activities and altered the cellular morphology of tumor cells. However, the effects of ethanol extract of baked GF (EBGF) on the metastatic and angiogenic capacities of malignant tumor cells and its detailed mechanism of action have not been reported. In this study, we found that EBGF significantly inhibited phorbol 12-myristate 13-acetate (PMA)-induced MMP-9 and -13 and uPA expression via suppression of PMA-induced nuclear translocation of NF-κBp65. Metastatic potential, including migration, invasion, and colonization, was substantially reduced by EBGF with no cytotoxicity. In addition, EBGF attenuated tumor-induced angiogenesis, including microvessel sprouting, migration of endothelial cells (ECs), and tube formation of ECs, by inhibiting the release of pro-angiogenic factors from tumor cells. In C57BL/6 mice, we observed that daily administration of EBGF at 50 and 100 mg/kg suppressed metastatic colonization of B16F10 melanoma cells in the lungs. Furthermore, EBGF administration did not cause adverse effects, suggesting that EBGF is safe and may be a potential herbal medicine capable of controlling metastatic malignant cancers.

System-level study on synergism and antagonism of active ingredients in traditional Chinese medicine by using molecular imprinting technology.

In this work, synergism and antagonism among active ingredients of traditional Chinese medicine (TCM) were studied at system-level by using molecular imprinting technology. Reduning Injection (RDNI), a TCM injection, was widely used to relieve fever caused by viral infection diseases in China. Molecularly imprinted polymers (MIPs) synthesized by sol-gel method were used to separate caffeic acid (CA) and analogues from RDNI without affecting other compounds. It can realize the preparative scale separation. The inhibitory effects of separated samples of RDNI and sample combinations in prostaglandin E2 biosynthesis in lipopolysaccharide-induced RAW264.7 cells were studied. The combination index was calculated to evaluate the synergism and antagonism. We found that components which had different scaffolds can produce synergistic anti-inflammatory effect inside and outside the RDNI. Components which had similar scaffolds exhibited the antagonistic effect, and the antagonistic effects among components could be reduced to some extent in RDNI system. The results indicated MIPs with the characteristics of specific adsorption ability and large scale preparation can be an effective approach to study the interaction mechanism among active ingredients of complex system such as TCM at system-level. And this work would provide a new idea to study the interactions among active ingredients of TCM.

Chemical fingerprinting of Gardenia jasminoides fruit using direct sample introduction and gas chromatography with mass spectrometry detection.

A rapid, rugged, and inexpensive approach is described to develop chemical fingerprints of volatile and semivolatile fractions in herbal medicine. The method is based on the combination of direct sample introduction and gas chromatography (GC) analysis with mass spectrometry detection. In comparison with routine methods, the proposed approach provides the most informative fingerprint and does not demand time-consuming extraction, pretreatment, and cleanup procedures. The approach was applied to establish the GC fingerprint of gardenia fruit (Gardenia jasminoides Ellis), in which 39 components were identified. With the help of principal components analysis, the obtained fingerprint could reveal the variation in and within different herb samples as affected by season and developmental state (wild or cultivated). The results indicated that the proposed approach could serve as a rapid, simple, and effective technique for the quality control of herbal medicines.