Geniposidic Acid Research Articles

O-methyltransferases catalyze the last step of geniposide biosynthesis in Gardenia jasminoides

Iridoid glycosides, including geniposide, geniposidic acid, and gardenoside, are the main active components of Gardenia jasminoides Ellis. Their biosynthetic pathways have widely attracted researchers’ attention, but yet remain unclear. Elucidating geniposide biosynthesis would help to better define the quality of G. jasminoides plants. In this study, we performed multidimensional transcriptional and metabolic analyses of the leaves and flower buds of G. jasminoides at different developmental stages based on the chromosome-level genome. Ten putative genes involved in geniposide biosynthesis were screened based on phylogenetic analysis and the correlation of gene expression with geniposide accumulation. Two non-tandem geniposidic acid methyltransferase (GAMT) genes were cloned, and in vitro enzymatic reactions with the corresponding recombinant proteins revealed that both could catalyze the conversion of geniposidic acid into geniposide. Synteny analysis showed that these two GAMT genes were unique to G. jasminoides, suggesting the existence of a Gardenia-specific geniposide biosynthetic pathway. In summary, two novel species-specific O-methyltransferases that catalyze the last step of geniposide biosynthesis were identified. These findings will help clarify the biosynthetic pathway of iridoid glycosides and support G. jasminoides breeding.

Study on enhanced extraction and seasonal variation of secondary metabolites in Eucommia ulmoides leaves using deep eutectic solvents

It has been considered as a crucial field for the extraction of active ingredients from herbal medicine to use a green and efficient method in the medicinal and food industries. In recent years, deep eutectic solvents (DESs) have been obtaining increase attention in green chemistry area since its sustainability, safety and biodegradability. In this study, an efficient DES composed of choline chloride and L-(+)-ascorbic acid with a molar ratio of 2:1 performed higher efficacy on the extraction of target compounds (including iridoids, phenolic acids and flavonoids) in Eucommia ulmoides leaves than 50% methanol solution. Considering the extraction efficacy and time consumption, microwave-assisted extraction (MAE) was selected and the operational conditions, including power of microwave, liquid/solid ratio and irradiation time were optimized by response surface methodology (RSM). Water was used as anti-solvent to recover ten target analytes from DES with recovery yields of 97.59%, 94.91%, 96.09%, 90.66%, 95.16%, 87.33%, 86.57%, 82.15%, 89.28% and 80.75% for eucommiol (EU), aucubin (AU), geniposidic acid (GA), chlorogenic acid (CA), asperuloside (AP), rutin (RU), kaempferol-3-O-sambubioside (KS), isoquercitrin (IQ), kaempferol-3-O-rutinoside (KR) and astragaline (AS), respectively. By combining the DES-based MAE and quantitative analysis of multi-components by single mark (QAMS) methods, the contents of ten compounds in the leaves of Eucommia ulmoides were determined to clarify the relationship between the accumulation of secondary metabolites and the harvest period. It was found that the contents of main ingredients reached the highest during May to October. The period appears to be the best harvest period for Eucommia ulmoides leaves. This study provides a novel strategy for the harvesting, processing, and quality control of the raw materials from Eucommia ulmoides leaves.

Inhibition Effect of <i>Eucommia ulmoides</i> Leaf Extract on Interleukin 8 Production by A549 Cells

Asthma is characterized by chronic inflammation of the airway mucosa. As Eucommia ulmoides Oliv. leaf extract (ELE) has been known to have anti-inflammatory properties, herein, we investigated the effect of ELE on interleukin (IL-) 8 production in A549 cells, a human airway epithelial cell line. The addition of ELE 1 h before tumor necrosis factor-alpha (TNFα) stimulation inhibited IL-8 production by A549 cells in a concentration-dependent manner. The addition of geniposidic acid, the main component of ELE, also inhibited IL-8 production. To further investigate the mechanism by which ELE inhibits IL-8 production, the effect of ELE or geniposidic acid on TNFα-stimulated p38 phosphorylation was examined by Western blotting. After 30 min of TNFα stimulation, p38 phosphorylation was inhibited by the addition of ELE or geniposidic acid, suggesting that ELE inhibited IL-8 production in TNFα-stimulated A549 cells by suppressing one of the signal transducers of p38 phosphorylation. These results indicate that ELE can be used as an effective measure against asthma, particularly neutrophilic asthma.

Deciphering the effects of genetic characteristics and environmental factors on pharmacological active ingredients of Eucommia ulmoides

The composition and yield of secondary metabolites of Eucommia ulmoides are important indices of the quality of medicinal materials, and their synthesis and accumulation are affected by both internal and external factors. However, the influential extent and mechanism of these factors on the pharmacologically active ingredients of E. ulmoides are still being elucidated. In this study, we selected 72 of E. ulmoides from 24 regions in China, and investigated the genetic diversity, pharmacologically active ingredients, soil physicochemical properties, and bark bacterial communities, along with the effects of genotypes and environmental factors on pharmacologically active ingredients. Results showed that genetic characteristics among the 24 regions there were no significant different, however, 12 different haplotypes were clearly revealed based on haplotype network analysis. The contents of pinoresinol diglucoside and geniposide acid were mainly affected by different climate and soil physiochemical factors, while aucubin was only sinificantly affected by soil organic matter. All of, the three pharmacologically active ingredients mentioned above were significantly correlated with the dominant genus Lactobacillus, Escherichia, Bacteroides and Faecalibacterium, suggesting that the active ingredients were affected by bark microorganisms. In conclusion, our analysis indicated that the accumulation of pharmacologically active ingredients of E. ulmoides were comprehensively affected genotype, environmental factors (biological and abiotic factors), and their interaction, with genotype had the greatest impact. These results provide important basic information for the future breeding, selection of cultivation conditions and assistance of microbial technology of E. ulmoides.

Rapid quality evaluation of Plantaginis Semen by near infrared spectroscopy combined with chemometrics

ObjectiveTo establish a rapid and comprehensive method for the quality evaluation of Plantaginis Semen (PS) by using near-infrared spectroscopy combined with chemometrics to determine the content of geniposidic acid (GEA), verbascoside (VE), isoverbascoside (IVE) and total flavonoids (TF) in PS, as well as the antioxidant activity (AOA). MethodsThe content of GEA, VE and IVE in PS were determined by HPLC, the content of TF in PS was determined by UV-Vis spectrophotometry, and the AOA of PS was characterized by the DPPH, ABTS and FRAP, respectively. Then, the measured values of each item were used as reference values and were correlated with the near infrared spectra of PS. Seven quantitative models were established by the partial least squares regression. A variety of spectral preprocessing, such as standard normal variation (SNV), multiplicative scatter correction (MSC), Savitzky-Golay smoothing (SG), derivative and their combination methods, were investigated. In addition, genetic algorithm (GA), particle swarm optimization (PSO), as well as competitive adaptive reweighted sampling (CARS) were also compared. All samples were divided into a calibration set and a prediction set at the ratio of 3:1 by the descending order of reference values. The coefficients of determination (R2), root mean square error (RMSE) and residual predictive deviation (RPD) were calculated to evaluate model performance. ResultsAfter optimization, the performance of each model was greatly improved, where the R2 for calibration and prediction were both greater than 0.8, the RPD were both greater than 2. Such satisfactory results indicated that the present models had good prediction accuracy. ConclusionQuantitative models based on near infrared spectroscopy were herein established, which proved to be able to quickly and accurately determine the content of GEA, VE and IVE and TF, as well as AOA in PS, and which might provide a new method for rapid and comprehensive quality evaluation of PS.

The influence of essential oils from ZhaLi NuSi Prescription on the pharmacokinetics of its non-volatile components in normal rats.

Hui Medicine ZhaLi NuSi Prescription (ZLNS) is described in "Hui Hui Prescription," and it has been used to treat cerebral infarction in Hui Region, China. In this study, a rapid and reliable ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS/MS) method was established and applied to simultaneously determine geniposidic acid, oxypaeoniflorin, hydroxysafflor yellow A, caffeic acid, magnoflorine, paeoniflorin, ferulic acid, β-ecdysterone, icariin, rhein, and baohuoside I in rat plasma. The pharmacokinetic parameters of these components and the influence of essential oils (EOs) on them were investigated in normal rats. The results showed that the pharmacokinetic parameters (AUC0 - t , AUC0 - ∞ , t1/2 , tmax , cmax ) of the aforementioned compounds were significantly changed after co-administering with ZLNS EO. The AUC values of oxypaeoniflorin, paeoniflorin, ferulic acid, and baohuoside I with EOs were decreased significantly. This is the first report for the comparative pharmacokinetic study of ZLNS bioactive components in normal rats, which may provide the basis for drug interaction study in vivo and insight into their clinical applications.

Effect of Tochu Extract and Its Component Geniposidic Acid on Renal Hemodynamics and Hypertensive Renal Damage

Effect of Tochu Extract and Its Component Geniposidic Acid on Renal Hemodynamics and Hypertensive Renal Damage

Effects of Geniposide and Geniposidic Acid on Fluoxetine-Induced Muscle Atrophy in C2C12 Cells

Fluoxetine, an antidepressant known as a selective 5-hydroxytryptamine reuptake inhibitor (SSRI), can cause side effects such as muscle atrophy with long-term use, but the mechanism is not fully understood. Geniposide (GPS) and geniposidic acid (GPSA), the main components of Gardenia jasminoides fruit, have been shown to have biological activity in disease prevention, but their role in preventing FXT-related side effects such as muscle atrophy remains unclear. The process of muscle atrophy is a complex physiological mechanism involving the balance of protein synthesis and catabolism. In this study, we hypothesized that FXT may suppress hypertrophy signaling and activate the atrophy mechanisms, resulting in proteolysis and reduced protein synthesis, while geniposide (GPS) and geniposide acid (GPSA) may be beneficial in improving muscle weakness caused by FXT. The C2C12 cell model was used to examine the expression of hypertrophy signaling (PI3K, Akt, and mTOR) and protein break signals (FOXO, MuRF-1, and MyHC). Our data indicated that FXT inhibited MyHC and promoted MuRF-1 protein expression by downregulating the signaling pathways of p-ERK1/2, p-Akt, p-mTOR, and p-FOXO, resulting in a decrease in differentiation and myotube formation in C2C12 muscle cells, which further resulted in muscle atrophy. However, GPS and GPSA can positively regulate the atrophy mechanism induced by FXT in muscle cells, thereby ameliorating the imbalance in muscle synthesis. In conclusion, GPS and GPSA have the potential to attenuate the muscle loss caused by long-term FXT administration, diseases, or the aging process.

Integrated Phytochemical Analysis Based on UPLC-MS/MS and Network Pharmacology Approaches to Explore the Effect of Odontites vulgaris Moench on Rheumatoid Arthritis.

Odontites vulgaris Moench has the effect of clearing away heat, detoxification, dispelling wind, and clearing dampness. In this study, the potential anti-inflammatory compounds of O. vulgaris were investigated using ultra-high-performance liquid chromatography–mass spectrometry (UPLC–MS/MS) combined with the network pharmacology approach and further confirmed on an LPS-activated RAW 264.7 macrophage model. Monomer compounds were prepared from the active fraction using modern advanced separation and purification methods. UPLC–Q-Exactive HRMS was used to identify the chemical compounds in the active fractions of O. vulgaris. D-mannitol, geniposidic acid, salidroside, shanzhiside methyl ester, eleutheroside B, geniposide, 7,8-dihydroxycoumarin, gardoside methyl ester, arenarioside, vanillic acid, p-hydroxy-cinnamic acid, melampyroside, syringaresinol, tricin, and diosmetin were isolated from O. vulgaris for the first time. A compound database of O. vulgaris was established based on the existing literature to predict the mechanism of O. vulgaris in the treatment of rheumatoid arthritis. The results suggest that the PI3K-Akt pathway mediates O. vulgaris and deserves more attention in the treatment of RA. Finally, the anti-rheumatoid arthritis effects of the four target compounds were validated with the decreased levels of NO, TNF-α, IL-6 and IL-1β in RAW 264.7 macrophage cells treated with LPS. The present study explored the potential targets and signaling pathways of O. vulgaris in the treatment of RA, which may help to illustrate the mechanisms involved in the action of O. vulgaris and may provide a better understanding of the relationship between O. vulgaris and RA. This study provides novel insights into the development of new drugs and utilization of Mongolian traditional Chinese medicine resources.

Phytosterol profiles and iridoids of the edible Eucommia ulmoides Oliver seeds and their anti-inflammatory potential

Eucommia ulmoides (EU) Oliver seeds are rich in oil and serve as a potential edible oil, but little is known about its phytochemical substances. In this study, the sterol profiles of the supercritical CO2 fluid extracted EU seed oil and its contents were first evaluated by GC-MS analysis, and fifteen sterols and three triterpene alcohols were determined. β-sitosterol was detected as the most abundant phytosterol in the seed oil. Additionally, seven iridoid glycosides (1–7), namely aucubin (1), bartsioside (2), linaride (3), geniposidic acid (4), one dimer scyphiphin D (5), two trimers ulmoidoside A (6) and ulmoidoside B (7) were identified from the defatted seed flour, and the compound 3 was reported in E. ulmoides for the first time. Natural and two acetylated compounds (1–9) were evaluated for their inhibitory effects on NO production in the lipopolysaccharide (LPS)-stimulated microglial cells, and 1–3 exerted significant inhibition of NO generation with IC50 values of 3.60, 7.09, and 10.52 μM, respectively. The structure-activity relationships (SARs) of these iridoids indicated that OH groups on C-10 and C-6 were more effective and the COOH group at C-4 position was highly detrimental to the activity. The results suggest that the edible EU seeds could be a potential dietary source of aucubin and β-sitosterol for reducing the risk of neurodegenerative diseases.

Drug-interaction between paclitaxel and goshajinkigan extract and its constituents

Paclitaxel, a standard chemotherapeutic agent for several types of cancer, including ovarian, breast, and non-small-cell lung cancer, causes peripheral neuropathy as an adverse effect in 60–70% of the patients. The utility of combination therapy with paclitaxel and goshajinkigan, a traditional Japanese Kampo medicine, in managing paclitaxel-induced neuropathy during chemotherapy has been explored. Paclitaxel is predominantly metabolized in the liver by cytochrome P450 (CYP) 2C8 to produce 6α-hydroxypaclitaxel and by CYP3A4 to produce 3′-p-hydroxypaclitaxel. In this study, we evaluated the inhibitory or inducing effects of goshajinkigan extract (GJG) and its representative and bioavailable constituents, geniposidic acid, plantagoguanidinic acid, paeoniflorin, catalpol, loganin, and neoline, on the metabolism of paclitaxel via CYP2C8 and CYP3A4 using pooled human liver microsomes and cultured human cryopreserved hepatocytes to provide the drug information about the pharmacokinetic interaction of this combination therapy. GJG significantly inhibited the production of 3’-p-hydroxypaclitaxel and 6α-hydroxypaclitaxel in vitro in a concentration-dependent manner. The half maximal inhibitory concentration (IC50) values of GJG were 4.5 and 7.8 mg/ml, respectively, for 3′-p-hydroxypaclitaxel and 6α-hydroxypaclitaxel productions. Neoline inhibited the production of 3′-p-hydroxypaclitaxel at 50 μM, but not at lower concentrations. Apart from neoline, other GJG constituents (at concentrations up to 50 or 10 μM of all test substances) did not exhibit inhibitory or inducing effects. Since GJG showed the inhibitory effect on the metabolism of paclitaxel at much higher concentrations than those used clinically, it can be concluded that GJG product does not exhibit any pharmacokinetic interaction with paclitaxel in clinical practice.Graphic abstract

Eucommia ulmoides (Tochu) and its extract geniposidic acid reduced blood pressure and improved renal hemodynamics

IntroductionEucommia ulmoides leaves are used as Tochu tea, which has a blood pressure lowering effect of unknown mechanism. Purpose and MethodsThe effects of Tochu tea and its component, geniposidic acid, on blood pressure and renal hemodynamics were investigated in Dahl salt-sensitive (DS) rats received 1% saline solution from 4 weeks of age. At 9 weeks of age, 1% saline alone (DSHS), Tochu tea extract added 1% saline (DSHS+T), or geniposidic acid added 1% saline (DSHS+G) was administered for another 4 weeks. DS rats fed with tap water were used as controls (DSLS). At 13 weeks, the blood pressure, the renal plasma flow (RPF) and the renal NADPH oxidase, endothelial nitric oxide synthase (eNOS) were examined. ResultsBlood pressure in DSHS rats was significantly increased in comparison to DSLS (144 vs. 196 mmHg, p < 0.01), and was significantly reduced in DSHS+T (158 mmHg) and DSHS+G (162 mmHg) rats. RPF in DSHS+T rats was significantly higher than in DSHS rats (p < 0.05). The expression of NADPH oxidase in DSHS rats was enhanced in comparison to DSLS rats; however, it was suppressed in DSHS+T and DSHS+G rats, and the NO production by eNOS was increased; thus, RPF was improved. The urinary Na excretion in DSHS rats was higher than that in DSLS rats; however it was further increased in DSHS+T rats without changes in the tubular Na transporters. ConclusionTochu tea and geniposidic acid suppressed NADPH oxidase, increased eNOS, and improved blood pressure and renal hemodynamics.

Extraction of geniposidic acid and aucubin employing aqueous two-phase systems comprising ionic liquids and salts

Ionic liquid aqueous two-phase systems (ILATPS) exhibiting enhanced separation efficiency and selectivity have attracted enormous attention. In this study, diverse ILATPSs comprising 1-alkyl-3-methylimidazolium ([Cnmim]+) and inorganic salts were developed and studied for the extraction of geniposidic acid (GPA) and aucubin (AU) for the first time. The binodal curves were fitted to the nonlinear Merchuk relationship. The ability of these ILs to form ATPS increased as their alkyl side chains increased. Moreover, the performances of the utilized potential salts in the phase separation were compared. The influences of the types and amounts of ILs and salts, extraction temperature and time, and the amount of the initial mixed solution were investigated through single-factor experiments. Under the following optimum conditions: 25% (w/w) K2CO3, 30% (w/w) [Bmim]Br, 1.2 mL of the GPA and AU mixed solution, temperature of 10 °C and extraction time of 360 min, the maximum extraction efficiency (E1AU = 96.95%, E1GPA = 99.99%), partition coefficient (logKAU = 1.54 ± 0.01 and logKGPA = 4.47 ± 0.01), and S (843.57 ± 15.67) were obtained. Afterward, IL could be easily recycled without organic solvents via a metathesis reaction with lithium bis(trifluoromethane)sulfonamide (LiNTf2) during which hydrophilic [Bmim]Br could be converted into hydrophobic [Bmim]NTf2. Furthermore, the density functional theory (DFT) calculations revealed that the H-bond interactions were the main factor that affected this extraction; the H-bond between [Bmim]Br and GPA was stronger than that between [Bmim]Br and AU. This research is expected to avail meaningful theoretical references for the extraction of similar natural products from plants.

A New Geniposidic Acid Derivative Exerts Antiaging Effects through Antioxidative Stress and Autophagy Induction.

Two compounds that can prolong the replicative lifespan of yeast, geniposidic acid (Compound 1) and geniposide (Compound 2), were isolated from Gardenia jasminoides Ellis. Compared with Compound 1, Compound 2 was different at C11 and showed better bioactivity. On this basis, seven new geniposidic derivatives (3–9) were synthesized. Geniposidic 4-isoamyl ester (8, GENI), which remarkably prolonged the replicative and chronological lifespans of K6001 yeast at 1 µM, was used as the lead compound. Autophagy and antioxidative stress were examined to clarify the antiaging mechanism of GENI. GENI increased the enzymes activities and gene expression levels of superoxide dismutase (SOD) and reduced the contents of reactive oxygen species (ROS) and malondialdehyde (MDA) to improve the survival rate of yeast under oxidative stress. In addition, GENI did not extend the replicative lifespan of ∆sod1, ∆sod2, ∆uth1, ∆skn7, ∆cat, and ∆gpx mutants with K6001 background. The free green fluorescent protein (GFP) signal from the cleavage of GFP-Atg8 was increased by GENI. The protein level of free GFP showed a considerable increase and was time-dependent. Furthermore, GENI failed to extend the replicative lifespans of ∆atg32 and ∆atg2 yeast mutants. These results indicated that antioxidative stress and autophagy induction were involved in the antiaging effect of GENI.

Metabolomic and Lipidomic Approaches to Evaluate the Effects of Eucommia ulmoides Leaves on Milk Quality and Biochemical Properties.

Eucommia ulmoides leaves (EUL) contain a variety of natural bioactive compounds including chlorogenic acid, geniposide acid, and aucubin. These bioactive chemicals improve immune function and regulate lipid metabolism. The aim of this study was to investigate the effects of EUL on the biochemical properties of milk. Twenty Holstein dairy cows were randomly allocated to two groups fed a control (CTR, diet without EUL, n = 10) or EUL (diet containing 3% EUL, dry matter, n = 10) diet for 55 d. At the end of the experimental period (d 55), milk samples were collected and analyzed to determine their composition. Though levels of milk fat, protein, lactose, and total milk solids were similar between the groups, small molecules, metabolites, lipids, and cytokines differed. Compared with the CTR group, the EUL group had an improved cluster of differentiation (CD)4/CD8 ratio (P < 0.05) and lower interleukin (IL)-8 and IL-6 content (P < 0.05). Metabolomics analysis identified 14 metabolites including 7Z, 10Z, 13Z, 16Z, 19Z-docosapentaenoic acid (FC = 3.129), adrenic acid (FC = 2.830), and eicosapentaenoic acid (FC=1.685) as having significantly increased in the EUL group (P < 0.05) while 11 metabolites, including indole-2-carboxylic acid (FC = 0.636), cholic acid (FC = 0.430), and creatine (FC = 0.784) had significantly decreased (P < 0.05). Based on a constructed metabolome map, linoleic acid metabolism had the highest impact value for EUL. A total of 21 lipid classes and 1,094 lipid species were detected in the milk by lipidomic analysis, among which 40 differed significantly between the CTR and EUL groups. The present findings showed that the EUL altered milk composition. Correlation analysis showed that 7Z, 10Z, 13Z, 16Z, 19Z-docosapentaenoic acid, adrenic acid, and eicosapentaenoic acid levels were negatively correlated with those of the inflammatory factors IL-6 and IL-8 (P < 0.05), indicating that EUL improved milk quality by reducing inflammatory factors and increasing the CD4/CD8 ratio. Overall, our data demonstrate that EUL had positive effects on milk antioxidant parameters, immune indices, and micro-composition metabolism, thereby improving milk quality.

A Combination of Geniposide and Chlorogenic Acid Combination Ameliorates Nonalcoholic Steatohepatitis in Mice by Inhibiting Kupffer Cell Activation.

The incidence of nonalcoholic steatohepatitis (NASH) is increasing worldwide. Activation of Kupffer cells (KCs) is central to the development of diet-induced NASH. We investigated whether a combination of two active chemical components, geniposide and chlorogenic acid (GC), at a specific ratio (67 : 1), ameliorates diet-induced NASH and the underlying mechanisms involved. C57BL/6J mice exposed to a high-fat and high-cholesterol (HFHC) diet containing cholesterol, choline, and high-sugar drinking water, as well as RAW264.7 cells stimulated with lipopolysaccharide (LPS) were studied. The combination exerted a therapeutic effect on HFHC-induced NASH in mice. Simultaneously, GC was found to reduce the expression of cytokines secreted by hepatic macrophages, including tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), IL-1β, IL-6, monocyte chemotactic protein 1 (MCP-1), and granulocyte-macrophage colony-stimulating factor (GM-CSF). Moreover, GC reduced the number of KCs expressing F4/80. Furthermore, TNF-α, inducible nitric oxide synthase (INOS), IL-1β, and IL-6 mRNA and TNF-α protein expression levels were suppressed upon GC treatment in RAW264.7 cells. Our findings suggest that GC has a strong anti-inflammatory effect in NASH, and this effect can be attributed to the suppression of KC activity in the liver.

Geniposide and Chlorogenic Acid Combination Improves Non-Alcoholic Fatty Liver Disease Involving the Potent Suppression of Elevated Hepatic SCD-1.

Background: Non-alcoholic fatty liver disease (NAFLD), characterized by the excessive accumulation of hepatic triglycerides (TGs), has become a worldwide chronic liver disease. But efficient therapy keeps unsettled. Our previous works show that geniposide and chlorogenic acid combination (namely the GC combination), two active chemical components combined with a unique ratio (67.16:1), presents beneficial effects on high-fat diet-induced NAFLD rodent models. Notably, microarray highlighted the more than 5-fold down-regulated SCD-1 gene in the GC combination group. SCD-1 is an essential lipogenic protein for monounsaturated fatty acids’ biosynthesis and serves as a key regulatory enzyme in the last stage of hepatic de novo lipogenesis (DNL). Methods: NAFLD mice model was fed with 16 weeks high-fat diet (HFD). The pharmacological effects, primarily on hepatic TG, TC, FFA, and liver enzymes, et al. of the GC combination and two individual components were evaluated. Furthermore, hepatic SCD-1 expression was quantified with qRT-PCR, immunoblotting, and immunohistochemistry. Finally, the lentivirus-mediated over-expressed cell was carried out to confirm the GC combination’s influence on SCD-1. Results: The GC combination could significantly reduce hepatic TG, TC, and FFA in NAFLD rodents. Notably, the GC combination presented synergetic therapeutic effects, compared with two components, on normalizing murine hepatic lipid deposition and disordered liver enzymes (ALT and AST). Meanwhile, the robust SCD-1 induction induced by HFD and FFA in rodents and ALM-12 cells was profoundly blunted, and this potent suppression was recapitulated in lentivirus-mediated SCD-1 over-expressed cells. Conclusion: Taken together, our data prove that the GC combination shows a substantial and synergetic anti-lipogenesis effect in treating NAFLD, and these amelioration effects are highly associated with the potent suppressed hepatic SCD-1 and a blunted DNL process.

Quality changes of Gardeniae Fructus Praeparatus processed by different frying methods: a color-component correlation analysis

The chromatic values of the broken-fried and single-fried Gardeniae Fructus Praeparatus(GFP) were measured by the color analyzer to analyze the color variation rule, and the contents of 10 main components were determined by ultra-high performance liquid chromatography(UPLC). The multivariate statistical analysis, Pearson correlation analysis, and discriminant analysis were conducted to investigate the color and components of GFP samples. The experimental results revealed that L~*, a~*, b~*, and E~*ab decreased continuously during processing, and the color of samples gradually deepened. The trend and range of chromatic values during broken-frying and single-frying processes were basically identical. Gardenoside, crocin-Ⅰ(C-Ⅰ), and crocin-Ⅱ(C-Ⅱ) showed an obviously downward trend, while the contents of geniposidic acid and 5-hydroxymethylfurfural(5-HMF) increased significantly. Shanzhiside, deacetyl-asperulosidic acid methyl ester, and geniposide(G2) showed a downward trend. Scandoside methyl ester rose first and fell later. Genipin-1-O-gentiobioside(G1) went through a decrease-increase-decrease trend. The change trends of component contents during broken-frying and single-frying processes were generally consistent, but the change range was different. Among all the components, scandoside methyl ester and G1 showed obvious change. Because of different stir-frying time, the change rate of each component content in the process of broken-frying was higher than that in single-frying process. Additionally, geniposidic acid, gardenoside, scandoside methyl ester, C-Ⅰ, C-Ⅱ, and 5-HMF exhibited a higher correlation with apparent color. On the basis of above findings, the discriminant function of two frying processes was established, which could be applied to the discrimination of broken-fried and single-fried samples. This study analyzed the dynamic quality change rule of GFP during broken-frying and single-frying processes based on color-component correlation analysis, and found the two methods showed consistent change trend, yet with slight difference in the quality of samples. This study can provide data support for the processing of GFP.

Analysis of blood components of Yougui Yin in normal rats and rats with kidney deficiency caused by adenine based on UPLC-MS technology

Based on the serum medicinal method, this study aims to investigate the migrating components of Yougui Yin in the blood after intragastric administration, and to provide reference for the basic research of its pharmacodynamics. The kidney deficiency rat model was replicated by adenine method. Normal rats and model rats were administered orally for a single gavage of Yougui Yin. The components in blood were rapidly analyzed and identified by ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and multiple reaction monitoring(MRM), and the migrating components in blood of Yougui Yin were explored by multivariate statistical analysis. The results showed that there were 42 characteristic peaks in the plasma of normal rats by UPLC-Q-TOF-MS technology and 13 chemical components were identified, including 6 alkaloids, 2 flavonoids, 2 triterpenoid saponins, 1 iridoid, 1 phenylpropanoid and 1 monoterpenoid. There were 22 characteristic peaks in the plasma of kidney-deficiency rats, and 12 chemical components were identified, including 2 iridoids, 6 alkaloids, 2 flavonoids, 1 monoterpenoid and 1 triterpenoid saponin. Verbascoside, isoacteoside, acteoside, pinoresinoldiglucoside, loganin and morroniside were identified by MRM both in the plasma of normal rats and kidney-deficiency rats. Compared with 85 monomer components in Yougui Yin, 17 common prototype components were found by UPLC-MS in the plasma of normal rats and kidney deficiency rats, including verbascoside, isoacteoside, acteoside, rehmapicrogenin derived from Rehmanniae Radix Praeparata, pinoresinol diglucoside and geniposidic acid from Eucommiea Cortex, loganin and morroniside derived from Corni Fructus, mesaconine, benzoylmesaconine, benzoylaconitine, benzoylhypacoitine, mesaconitine, aconitine derived from Aconiti Lateralis Radix Praeparata, liquiritin, isoliquiritin and glycyrrhizic acid derived from Glycyrrhizae Radix et Rhizoma. Thirty-one metabolites of medicinal ingredients not found in the plasma of adenine-induced kidney deficiency rats were also detected in the plasma of normal rats. Twelve metabolites of medicinal materials not found in the plasma of normal rats were detected in the plasma of kidney deficiency rats. The results of the study provide reference for explaining the material basis and mechanism of Yougui Yin in the treatment of kidney deficiency.

Simultaneous quantification of bioactive components in Chinese herbal spirits by ultra-high performance liquid chromatography coupled to triple-quadrupole mass spectrometry (UHPLC\u2013QQQ\u2013MS/MS)

BackgroundThe Chinese medicinal wine made from herbal medicines became prevalent among Chinese people. The Chinese herbal spirit is composed of several herbal extracts, and has the certain health functions, such as anti-fatigue and immune regulation. The quality evaluation of Chinese herbal spirit is greatly challenged by the enormous and complex components with great structural diversity and wide range of concentration distribution.MethodsAn ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) with multiple reaction monitoring (MRM) method was developed to simultaneously determine forty-three bioactive components in the Chinese herbal spirits produced by year 2014 and 2018.ResultsQuantitative results showed that 11 components, i.e.., puerarin (5), purpureaside C (7), daidzin (8), echinacoside (9), acteoside (15), epimedin B (22), epimedin C (23), icariin (24), eugenol (27), chikusetsusaponin iva (30) and Z-ligustilide (40), significantly decreased along with the increasing years of storage, while 5 compounds, i.e.., geniposidic acid (1), protocatechuic acid (2), crustecdysone (14), daidzein (18) and icariside I (35), were basically stable in all samples across the years.ConcusionThe established method allowing to simultaneously determined 43 components with wide structural diversity and trace amounts will facilitate the quality control research of Chinese herbal spirits.