Glycyrrhetinic Acid Research Articles

Integration of serum pharmacochemistry and metabolomics to reveal the underlying mechanism of shaoyao-gancao-fuzi decoction to ameliorate rheumatoid arthritis

Ethnopharmacological relevanceFor centuries, Shaoyao-Gancao-Fuzi decoction (SGFD) has been a reliable traditional Chinese medicine for treating rheumatoid arthritis (RA). Despite its long history of use, the specific active components and underlying mechanisms of its therapeutic effects have yet to be fully understood. Aim of the studyThe aim of this study was to investigate the active ingredients and therapeutic effects of SGFD on RA, and to further understand its underlying mechanism. Materials and methodsThe chemical constituents in SGFD extract and in rat serum after oral administration of SGFD were identified and evaluated using ultra-performance liquid chromatography quadrupole-time-flight mass spectrometry (UPLC-Q-TOF/MS) together with various data-processing methods, respectively. The efficacy of SGFD was assessed by using an adjuvant-induced arthritis (AIA) rat model and lipopolysaccharide-stimulated RAW 264.7 cell. Subsequently, cell metabolomic was conducted to clarify the potential biomarkers and pathways. ELISA, RT-qPCR, and WB were used to verify the anti-arthritis mechanism of SGFD. ResultsA total of 65 chemical constituents were identified in SGFD. 17 active components were distinguished in rat serum samples, of which 13 may be the main active ingredients for SGFD treatment of RA. The remarkable efficacy of SGFD in reducing the symptoms of RA is evident through its ability to alleviate the redness and swelling of the affected paws, as well as reduce the infiltration of inflammatory cells. Cell experiments revealed that rat serum of SGFD reduced IL-1β, IL-6, and TNF-α secretion in RAW 264.7 cells. 27 potential biomarkers were identified through cell metabolomics analysis. The arachidonic acid (AA) metabolism signaling pathway was activated in RA, which could be reversed by rat serum of SGFD. SGFD effectively inhibited the expression and transformation of AA by downregulating the expression of key enzymes, including phospholipase A and cyclooxygenase. ConclusionSGFD may ameliorate RA symptoms by regulating the AA-PGH2-PGE2/PGF2α pathway. The main active components include songorine, fuziline, neoline, albiflorin, paeoniflorin, liquiritin, benzoylmesaconine, isoformononetin, liquiritigenin, isoliquiritigenin, formononetin, glycyrrhizic acid, and glycyrrhetinic acid.

Natural Sweetener Glycyrrhetinic Acid Monoglucuronide Improves Glucose Homeostasis in Healthy Mice.

Noncaloric or low-caloric sweeteners have become popular worldwide, although debates persist regarding their impact on health. To investigate whether the sweeteners are favorable for glucose homeostasis, our study assessed the effects of glycyrrhetinic acid monoglucuronide (GAMG) and several commonly used sweeteners [glycyrrhetinic acid (GA), stevioside, erythritol, sucralose, and aspartame] on glycometabolism and elucidated the underlying mechanisms. The C57BL/6J male mice were exposed to different sweeteners for 10 weeks, and our results showed that GAMG significantly reduced fasting blood glucose (FBG) levels (FBG-control: 3.81 ± 0.42 mmol/L; FBG-GAMG: 3.37 ± 0.38 mmol/L; p < 0.05) and the blood glucose levels 15 and 30 min after sucrose or maltose loading (p < 0.05). Furthermore, it improved glucose tolerance (p = 0.028) and enhanced insulin sensitivity (p = 0.044), while the other sweeteners had negligible or adverse effects on glucose homeostasis. Subsequent experiments showed that GAMG inhibited α-glucosidases potently (IC50 = 0.879 mg·mL-1), increased three SCFA-producing bacteria and SCFAs levels (p < 0.05), and promoted the gene expression of SCFA receptor GPR43 (p = 0.018). These results suggest that GAMG may regulate blood glucose by inhibiting α-glucosidases and modulating gut microbial SCFAs. Our findings prove that GAMG, beneficial to blood glucose regulation, is a promising natural sweetener for future utilization.

Natural Sweetener, Glycyrrhetinic Acid 3-O-Mono-beta-d-glucuronide, for Postprandial Hyperglycemia Management.

This study examines the inhibitory effects of a range of sweeteners on α-glucosidase. Our findings revealed that only one natural sweetener, namely, glycyrrhetinic acid 3-O-mono-beta-d-glucuronide (GAMG), derived from licorice, exhibited a mixed-type inhibition against α-glucosidase with a IC50 value of 0.73 ± 0.05 mg/mL. The fluorescence intensity of α-glucosidase was quenched by GAMG in the formation of an α-glucosidase-GAMG complex. GAMG has been shown to induce conformational changes in α-glucosidase, likely through hydrogen bonding, van der Waals force, and alkyl-alkyl interactions with amino acid residues, including Arg 281, Leu 283, Trp 376, Asp 404, Asp 443, Trp 481, Asp 518, Phe 525, Ala 555, and Asp 616. Additional animal validation experiments demonstrated that GAMG slowed starch digestion, thereby attenuating the postprandial glycemic response. Taken together, these findings provide evidence that GAMG is a natural sweetener with potent inhibitory activity that selectively targets α-glucosidase. This study supports the use of GAMG as a natural sweetener, which holds a high biological value and may be beneficial for managing postprandial hyperglycemia.

Biosynthesis of Chryseno[2,1,c]oxepin-12-Carboxylic Acid from Glycyrrhizic Acid in Aspergillus terreus TMZ05-2, and Analysis of Its Anti-inflammatory Activity.

Glycyrrhizic acid, glycyrrhetinic acid, and their oxo, ester, lactone, and other derivatives, are known for their anti-inflammatory, anti-oxidant, and hypoglycemic pharmacological activities. In this study, chryseno[2,1-c]oxepin-12-carboxylic acid (MG) was first biosynthesized from glycyrrhizic acid through sequential hydrolysis, oxidation, and esterification using Aspergillus terreus TMZ05-2, providing a novel in vitro biosynthetic pathway for glycyrrhizic acid derivatives. Assessing the influence of fermentation conditions and variation of strains during culture under stress-induction strategies enhanced the final molar yield to 88.3% (5g/L glycyrrhizic acid). CCK8 assays showed no cytotoxicity and good cell proliferation, and anti-inflammatory experiments demonstrated strong inhibition of NO release (36.3%, low-dose MG vs. model), transcriptional downregulation of classical effective cellular factors tumor necrosis factor-α (TNF-α; 72.2%, low-dose MG vs. model), interleukin-6 (IL-6; 58.3%, low-dose MG vs. model) and interleukin-1β (IL-1β; 76.4%, low-dose MG vs. model), and decreased abundance of P-IKK-α, P-IKB-α, and P-P65 proteins, thereby alleviating inflammatory responses through the NF-κB pathway in LPS-induced RAW264.7 cells. The findings provide a reference for the biosynthesis of lactone compounds from medicinal plants.

Glycyrrhetinic acid alleviates atrial fibrillation by inhibiting the expression of PIM1

AbstractGlycyrrhetinic acid (GA) is known to exert a cardioprotective effect, while its mechanism in atrial fibrillation (AF) remains poorly defined. Accordingly, isoprenaline (ISO) was applied to establish the rat AF model, and then GA (5, 10, and 20 mg/kg/day) was administrated to these rats for a total of 21 days. The fibrosis and pathological conditions of rat atrial tissues were observed. Quantitative real‐time polymerase chain reaction (qRT‐PCR) was adopted to calculate the levels of proto‐oncogene serine/threonine‐protein kinase Pim1 (PIM1) and Interleukin (IL)‐4/6/13/17. For in‐vitro experiments, isolated cardiac fibroblasts received the transfection with PIM1‐specific overexpression plasmid or siPIM1 and the treatment with ISO. Subsequently, cell viability was gauged by cell counting kit‐8 assay. The protein levels of alpha‐smooth muscle actin (α‐SMA), collagen I and III, transforming growth factor‐beta1 (TGF‐β1) and mothers against decapentaplegic homolog 7 (Smad7) were measured by western blot. GA led to the shortened duration of AF and alleviated myocardial fibrosis and atrial tissue lesions in ISO‐treated AF rats in vivo. Meanwhile, GA inhibited the expressions of PIM1 and IL‐4/6/13/17 in the atrial tissue of AF model rats. In the meantime, enhanced cell viability, inhibited expressions of PIM1, IL‐4/6/13/17, α‐SMA, collagen‐I, collagen‐III and TGF‐β1, and promoted Smad7 expression were observable in ISO‐treated cardiac fibroblasts in vitro. Oppositely, the effects of GA were reversed following PIM1 overexpression, while knockdown of PIM1 enhanced the effects of GA on cardiac fibroblasts. Collectively speaking, GA alleviates AF through suppressing the expression of PIM1.

Low temperature promotes the production and efflux of terpenoids in yeast

Altering the fermentation environment provides an effective approach to optimizing the production efficiency of microbial cell factories globally. Here, lower fermentation temperatures of yeast were found to significantly improve the synthesis and efflux of terpenoids, including glycyrrhetinic acid (GA), β-caryophyllene, and α-amyrin. The production of GA at 22°C increased by 5.5 times compared to 30°C. Yeast subjected to lower temperature showed substantial changes at various omics levels. Certain genes involved in maintaining cellular homeostasis that were upregulated under the low temperature conditions, leading to enhanced GA production. Substituting Mvd1, a thermo-unstable enzyme in mevalonate pathway identified by transcriptome and proteome, with a thermo-tolerant isoenzyme effectively increased GA production. The lower temperature altered the composition of phospholipids and increased the unsaturation of fatty acid chains, which may influence GA efflux. This study presents a strategy for optimizing the fermentation process and identifying key targets of cell factories for terpenoid production.

A multistep approach for exploring quality markers of Shengjiang Xiexin decoction by integrating plasma pharmacochemistry-pharmacokinetics-pharmacology

Shengjiang Xiexin decoction (SXD), a well-known traditional Chinese medicine (TCM), was used to alleviate delayed-onset diarrhea induced by the chemotherapeutic agent irinotecan (CPT-11). Our previous study showed that SXD regulated multidrug resistance-associated protein 2 (Mrp-2) to alter the pharmacokinetics of CPT-11 and its metabolites. However, the pharmacodynamic constituents and the related quality markers of SXD are unclear. In this study, ultra-high performance liquid chromatography coupled with quadrupole orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was utilized to identify the prototypes and metabolites in rat plasma after oral administration of SXD. The pharmacokinetic markers (PK markers) were screened through quantification and semiquantification of SXD-related xenobiotics in plasma using liquid chromatography-mass spectrometry (LC-MS) combined with statistical analysis. Computational molecular docking was performed to assess the potential binding ability of the PK markers with the target Mrp-2. The results were verified by evaluating the impact on Mrp-2 function using Caco-2 cells. The quality markers were chosen from these PK markers based on the binding affinities with Mrp-2, the specificity and the traceability. As a result, a total of 142 SXD-related exogenous components, including 77 prototypes and 65 metabolites, were detected in rat plasma. Among these, 83 xenobiotics were selected as PK markers due to their satisfactory pharmacokinetic behaviors. Based on the characteristics of quality markers, the prototype-based PK markers were considered the indices of quality control for SXD, including baicalin, baicalein, wogonoside, wogonin, liquiritigenin, isoliquiritigenin, norwogonin, oroxylin A, dihydrobaicalin, chrysin, glycyrrhizic acid, glycyrrhetinic acid, oroxylin A 7-O-glucuronide, liquiritin and isoliquiritin. This study provided an interesting strategy for screening the quality markers involved in the pharmacokinetics of SXD and its action target, which offered important information for the modernization of SXD and other TCM formulae.

Two new compounds from biotransformation of glycyrrhetinic acid

Glycyrrhetinic acid may undergo biotransformation to obtain derivatives with stronger pharmacological activity and fewer side effects. This study used 19 fungi to biotransform glycyrrhetinic acid and yielded two compounds namely bicyclo(14.15.27)glycyrrhetinic acid (1) and 2-ene-glycyrrhetinic acid (2) from the glycyrrhetinic acid metabolites of two fungi, Botrytis cinerea B05.10 ATCC 11542 and Aspergillus ochraceopetaliformis ATCC 12066. Compound 1 inhibited HMEC-1 cell proliferation in a concentration-dependent manner (IC50=239.1 µ M), but showed no significant cytotoxicity to A549 cells.

Anti-tumor therapy of glycyrrhetinic acid targeted liposome co-delivery of doxorubicin and berberine for hepatocellular carcinoma.

During the development of hepatocellular carcinoma (HCC), hepatic stellate cells undergo activation and transform into cancer-associated fibroblasts (CAFs) due to the influence of tumor cells. The interaction between CAFs and tumor cells can compromise the effectiveness of chemotherapy drugs and promote tumor proliferation, invasion, and metastasis. This study explores the potential of glycyrrhetinic acid (GA)-modified liposomes (lip-GA) as a strategy for co-delivery of berberine (Ber) and doxorubicin (Dox) to treat HCC. The characterizations of liposomes, including particle size, zeta potential, polydispersity index, stability and in vitro drug release, were investigated. The study evaluated the anti-proliferation and anti-migration effects of Dox&Ber@lip-GA on the Huh-7 + LX-2 cell model were through MTT and wound-healing assays. Additionally, the in vivo drug distribution and anti-tumor efficacy were investigated using the H22 + NIH-3T3-bearing mouse model. The results indicated that Dox&Ber@lip-GA exhibited a nanoscale particle size, accumulated specifically in the tumor region, and was efficiently taken up by tumor cells. Compared to other groups, Dox&Ber@lip-GA demonstrated higher cytotoxicity and lower migration rates. Additionally, it significantly reduced the deposition of extracellular matrix (ECM) and inhibited tumor angiogenesis, thereby suppressing tumor growth. In conclusion, Dox&Ber@lip-GA exhibited superior anti-tumor effects both in vitro and in vivo, highlighting its potential as an effective therapeutic strategy for combating HCC.

Evaluation of Antioxidants’ Ability to Enhance Hyaluronic Acid-Based Topical Moisturizers

Introduction: Hyaluronic acid (HA) is a unique molecule with multiple biological activities. In skin, HA plays an essential role as a humectant, capable of binding up to 1000 times its mass with water, leading to both skin moisturization and extracellular matrix rigidity. HA concentration and synthesis decreases significantly in aging skin, due to exogenous and endogenous factors, including photoaging and HA metabolism, respectively. A key driver for HA degradation and decrease in concentration is mediated via the induction of reactive oxygen species (ROS) and other free radicals. In this study, we evaluate antioxidant ingredients essential in the development of next-generation multi-weight HA-based topical formulations aimed at leveraging hyaluronic acid’s ability to maximize anti-aging properties. Methods: Two antioxidants, glycine saponin and glycyrrhetinic acid, were evaluated for stimulation of endogenous HA production and inhibition of endogenous hyaluronidase activity in an in vitro and cell-free assay. Results: The antioxidant glycine saponin induced endogenous HA synthesis in keratinocytes, while the antioxidant glycyrrhetinic acid demonstrated an approximately 54% reduction of hyaluronidase activity for the HA-degrading enzyme HYAL1. Conclusion: This study demonstrates the ability of two antioxidants (glycine saponin and glycyrrhetinic acid, essential in the development of the next generation of multi-weight HA plus antioxidant complex-based topical skin formulations) to limit the signs of aging skin, addressing limitations of many currently available topical HA-based formulations.

Significantly Enhanced Improvement in Dryness, Roughness, Fine Lines and Radiance Following Daily Use of a Novel Multi-Weight Hyaluronic Acid Plus Antioxidant Complex-Based Lotion Compared to a Single-Weight HA Plus Ceramide-Based Lotion

Introduction: Hyaluronic acid (HA) has become a commonly used ingredient in many topical moisturizing products due to its strong humectant properties and essential role in skin hydration; however, limitations of delivery of HA to only the surface of skin has hindered leveraging the full capacity of HA biology necessary for skin rejuvenation. Here we describe the head-to-head clinical comparison of a novel multi-weight HA plus antioxidant complex-based lotion with SPF 30 and a single-weight plus ceramide-based lotion with SPF 30 for clinical efficacy on dryness, roughness, fine facial lines, and radiance following daily use. Methods: A double-blind comparative study was conducted on 70 female subjects (n=35, multi-weight HA plus antioxidant complex-based lotion with SPF 30; n=35, single-weight plus ceramide-based lotion with SPF 30) ages 25 to 65 years with mild to moderate facial dryness and visible fine lines and wrinkles, including subjects with Fitzpatrick Skin Types I-VI, with 20% having Fitzpatrick Skin Types V-VI. Clinical grading of the face including dryness, roughness, and fine lines were assessed at baseline, 30 minutes, 2 weeks, 4 weeks, and 8 weeks after once daily application in the morning. Visible, cross-polarized (X-Pol), parallel-polarized (P-Pol) and UV fluorescence clinical images were acquired for each time point. X-Pol and P-Pol images were used to quantify skin radiance. Results: Daily use of the multi-weight HA plus antioxidant complex-based lotion with SPF 30 demonstrated significant improvements in all clinical grading assessments of dryness, fine lines, and wrinkles as early as Week 2 compared to baseline. Improvements in visible dryness (Week 2), roughness (Week 2), and fine lines (Week 8) were greater for the multi-weight HA plus antioxidant complex-based lotion with SPF 30 compared to the single-weight HA plus ceramide-based lotion with SPF 30, with overall statistical significance across all three parameters assessed favoring the multi-weight HA plus antioxidant complex-based lotion with SPF 30. For radiance, daily use of the multi-weight HA plus antioxidant complex-based lotion with SPF 30 demonstrated significant increase in radiance at Week 4 compared with baseline, while the single-weight HA plus ceramide-based lotion did not show any improvement in skin radiance compared with baseline. Conclusions: The marked improvements in dryness, roughness, fine lines, and radiance following daily utilization of the multi-weight HA plus antioxidant complex-based lotion with SPF 30 may be attributed to the inherent properties of HA. These improvements may be further attributed to the ability of multi-weight HAs to moisturize the skin surface and penetrate the upper surface layers of the skin, combined with the added benefits of key antioxidants, including glycine saponin and glycyrrhetinic acid, which have been previously shown to induce endogenous HA synthesis and inhibit endogenous hyaluronidase activity in vitro, respectively.

Flubendazole carbonyl reduction in drug-susceptible and drug-resistant strains of the parasitic nematode Haemonchus contortus: changes during the life cycle and possible inhibition

Carbonyl-reducing enzymes (CREs) catalyse the reduction of carbonyl groups in many eobiotic and xenobiotic compounds in all organisms, including helminths. Previous studies have shown the important roles of CREs in the deactivation of several anthelmintic drugs (e.g., flubendazole and mebendazole) in adults infected with the parasitic nematode Haemonchus contortus, in which the activity of a CRE is increased in drug-resistant strains. The aim of the present study was to compare the abilities of nematodes of both a drug-susceptible strain (ISE) and a drug-resistant strain (IRE) to reduce the carbonyl group of flubendazole (FLU) in different developmental stages (eggs, L1/2 larvae, L3 larvae, and adults). In addition, the effects of selected CRE inhibitors (e.g., glycyrrhetinic acid, naringenin, silybin, luteolin, glyceraldehyde, and menadione) on the reduction of FLU were evaluated in vitro and ex vivo in H. contortus adults. The results showed that FLU was reduced by H. contortus in all developmental stages, with adult IRE females being the most metabolically active. Larvae (L1/2 and L3) and adult females of the IRE strain reduced FLU more effectively than those of the ISE strain. Data from the in vitro inhibition study (performed with cytosolic-like fractions of H. contortus adult homogenate) revealed that glycyrrhetinic acid, naringenin, mebendazole and menadione are effective inhibitors of FLU reduction. Ex vivo study data showed that menadione inhibited FLU reduction and also decreased the viability of H. contortus adults to a similar extent. Naringenin and mebendazole were not toxic at the concentrations tested, but they did not inhibit the reduction of FLU in adult worms ex vivo.

Structure–Activity relationship (SAR) of 18β-glycyrrhetinic acid derivatives on their antitumor activity through the PPARγ receptor and caspase-3 pathway

Licorice is extensively utilized in the food industry, and glycyrrhetinic acid is among the most critical active constituents in licorice. This ingredient has garnered increasing attention in recent years. In a previous study, we identified a new series of Glycyrrhetinic acid derivatives containing piperazine skeleton, using virtual screening techniques based on docking. In this study, we conducted further optimization of the lead compound 4c, with a focus on enhancing the hydrophobic interactions in the binding site to improve agonist efficacy for peroxisome proliferator-activated receptor γ (PPARγ). The obtained compounds were characterized using 1H NMR, 13C NMR, HRMS, and HPLC. Furthermore, these compounds were evaluated strong antitumor activity. Some compounds with potent antitumor activities were tested for their PPARγ agonist activity. Especially, compound 4f showed the most potent antitumor activity with half maximal inhibitory concentration (IC50) values of 2.382 μM against the tested tumor cell lines Siha. Docking simulation was performed to position compound 4f into the PPARγ active site to determine the probable binding conformation. Consequently, we discovered a novel PPARγ agonist 4f that demonstrated high in vitro agonist activity.

Retraction Note: One-step Conjugation of Glycyrrhetinic Acid to Cationic Polymers for High-performance Gene Delivery to Cultured Liver Cell

Retraction Note: One-step Conjugation of Glycyrrhetinic Acid to Cationic Polymers for High-performance Gene Delivery to Cultured Liver Cell

A review of typical biological activities of glycyrrhetinic acid and its derivatives.

Glycyrrhetinic acid, a triterpenoid compound primarily sourced from licorice root, exhibits noteworthy biological attributes, including anti-inflammatory, anti-tumor, antibacterial, antiviral, and antioxidant effects. Despite these commendable effects, its further advancement and application, especially in clinical use, have been hindered by its limited druggability, including challenges such as low solubility and bioavailability. To enhance its biological activity and pharmaceutical efficacy, numerous research studies focus on the structural modification, associated biological activity data, and underlying mechanisms of glycyrrhetinic acid and its derivatives. This review endeavors to systematically compile and organize glycyrrhetinic acid derivatives that have demonstrated outstanding biological activities over the preceding decade, delineating their molecular structures, biological effects, underlying mechanisms, and future prospects for assisting researchers in finding and designing novel glycyrrhetinic acid derivatives, foster the exploration of structure-activity relationships, and aid in the screening of potential candidate compounds.

Exploring the Role of Licorice and Its Derivatives in Cell Signaling Pathway NF-κB and MAPK.

Licorice is a therapeutic herb in traditional Chinese herbal medicine. Licorice is considered as an anti-inflammatory agent due to its suppression and inhibition of inflammatory pathways. Licorice has many bioactive compounds such as glycyrrhetinic acid, glycyrrhizin, liquiritigenin, and isoliquirtigenin which are principally accountable for its therapeutic benefits. These bioactive components reduce inflammation by preventing the activation of important inflammatory pathways including mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-κB). As a result of this tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) are among the proinflammatory cytokines whose production is inhibited. Components present in licorice inhibit the activation by suppressing the IκBα phosphorylation and degradation. Moreover, licorice compounds also attenuate the MAPK signaling cascades by inhibiting the MAPK kinase phosphorylation and downstream MAPKs such as extracellular signal-regulated kinase (ERK), p38 MAPK, and c-Jun N-terminal kinase (JNK). The present review focuses on the current understanding of licorice effect on the NF-κB and MAPK inflammatory cell signaling pathways at molecular level. Furthermore, emerging evidence suggested that licorice-derived bioactive compounds may attenuate the molecular mechanism which is associated with inflammation, providing the additional insights into the therapeutic potential. Further studies explained the precise molecular mechanism at the cellular level underlying the licorice anti-inflammatory effect and potential application in managing inflammatory disorders. In conclusion, licorice has a complex mode of action and is a valuable natural anti-inflammatory. Its natural origin and effectiveness in clinical applications make it an intriguing topic for additional study. As licorice becomes more widely used in medicine, future research should focus on refining its formulations to optimize therapeutic advantages.

Network pharmacology mechanisms and experimental verification of licorice in the treatment of ulcerative colitis

Ethnopharmacological relevanceLicorice is widely used in the treatment of ulcerative colitis (UC) and has good antioxidant and anti-inflammatory effects, but its specific active ingredients and mechanisms of action are still unknown. The purpose of the studyTo elucidate the specific molecular mechanisms of licorice in the treatment of UC and to experimentally verify its activity. MethodsThrough network pharmacology, the active ingredients of licorice and the molecular targets of UC were identified. A traditional Chinese medicine (TCM)-components-target-disease network diagram was established, and the binding energies of the active ingredient and targets of licorice were verified by molecular docking. A BALB/c mice model of UC was established by treatment with 3% dextran sulfate sodium (DSS). The effect of licorice on colon tissue injury was histologically assessed. The expression of IL-6 and IL-17 in colon tissue was detected by immunohistochemistry (IHC). Transmission electron microscopy (TEM) was used to observe morphological changes in mitochondria in the colon. Caco2 cells were treated with lipopolysaccharide (LPS) for 24 h to establish the cell inflammatory damage model, and cells were exposed to different concentrations of drug-containing serum of Licorice (DCSL) for 24 h. In cells treated with the drug, the contents of oxidation markers were measured and ELISA was used to determine the levels of inflammatory factors in the cells. TEM was used to observe morphological changes in mitochondria. ZO-1 and occludin were detected by Western blotting. DCSL effects on autophagy were evaluated by treating cells with DCSL and autophagy inhibitor for 24 h after LPS injection. Small interfering ribonucleic acid (si-RNA) was used to silence Nrf2 gene expression in Caco2 cells to observe the effects of DCSL on autophagy through the Nrf2/PINK1 pathway. Nrf2, PINK1, HO-1, Parkin, P62, and LC3 were detected by Western blotting. ResultsNinety-one active ingredients and 339 action targets and 792 UC disease targets were identified, 99 of which were overlapping targets. Molecular docking was used to analyze the binding energies of liquiritin, liquiritigenin, glycyrrhizic acid, and glycyrrhetinic acid to the targets, with glycyrrhetinic acid having the strongest binding energy. In the UC mouse model, licorice improved colon histopathological changes, reduced levels of IL-6 and IL-17 and repaired mitochondrial damage. In the LPS-induced inflammation model of Caco2 cells, DCSL decreased MDA, IL-1β, Il-6, and TNF-α levels and increased those of Superoxide Dismutase (SOD), glutathione peroxidase (GSH-PX), and IL-10, and improved the morphological changes of mitochondria. Increased expression of Nrf2, PINK1, Parkin, HO-1, ZO-1, occludin, P62, and LC3 promoted autophagy and reduced inflammation levels. ConclusionLicorice improves UC, which may be related to the activation of the Nrf2/PINK1 signaling pathway that regulates autophagy.

Chromatographic Quantitative Analysis of Diosgenin, Gallic Acid, and Glycyrrhetinic Acid

ABSTRACT Background: Amla, liquorice, and fenugreek have three primary bioactive phytoconstituents: diosgenin, gallic acid, and 18β-glycyrrhetic acid. This work discusses the invention and validation of an high-performance liquid chromatography (HPLC) approach for the simultaneous detection, in line with ICH criteria, of gallic acid in amla, glycyrrhetinic acid in liquorice, and diosgenin in fenugreek. Materials and Methods: A new simple, exact, sensitive, and validated reversed phase (RP)-HPLC technique was developed for the measurement of diosgenin, gallic acid, 18β-glycyrrhetic acid, and acid in bulk and pharmaceutical dosage form. Acetonitrile: 0.05% ortho phosphoric acid (OPA) (95:5) for diosgenin in fenugreek extract, a rheodyne manual injector with a 20 μl capacity, and a Phenomenex Luna C18 (2) (4.6 mm × 250 mm, 5) chromatographic apparatus were used for the separation. In Amla extract, methanol: 0.05% OPA (70:30) for gallic acid liquorice extract: Methanol: Acetonitrile: 0.05% OPA for 18β-glycyrrhetic acid A solution of ophosphoric acid (OPA) was used to maintain the pH at 3.00. The detection wavelengths utilized were 205 nm for diosgenin, 272 nm for gallic acid, and 250 nm for 18β-glycyrrhetic acid. The flow rate was set at 1.0 mL/min. Results: Even though HPLC is more sensitive for diosgenin analysis, the developed RP-HPLC method should offer a fast, accurate, simple, and inexpensive alternative approach for the quantitative detection of diosgenin, gallic acid, and 18β-glycyrrhetic acid. Conclusion: The process described in this paper produces the same amount of pure diosgenin as other reported methods.

Protective effect of glycyrrhetinic acid against cantharidin-induced hepatotoxicity through reducing oxidative stress in mice

Protective effect of glycyrrhetinic acid against cantharidin-induced hepatotoxicity through reducing oxidative stress in mice

Disruption of a licorice cellulose synthase-derived glycosyltransferase gene demonstrates its in planta role in soyasaponin biosynthesis

Key messageCRISPR–Cas9-mediated disruption of a licorice cellulose synthase-derived glycosyltransferase gene, GuCSyGT, demonstrated the in planta role of GuCSyGT as the enzyme catalyzing 3-O-glucuronosylation of triterpenoid aglycones in soyasaponin biosynthesis.Triterpenoid glycosides (saponins) are a large, structurally diverse group of specialized metabolites in plants, including the sweet saponin glycyrrhizin produced by licorice (Glycyrrhiza uralensis) and soyasaponins that occur widely in legumes, with various bioactivities. The triterpenoid saponin biosynthetic pathway involves the glycosylation of triterpenoid sapogenins (the non-sugar part of triterpenoid saponins) by glycosyltransferases (GTs), leading to diverse saponin structures. Previously, we identified a cellulose synthase-derived GT (CSyGT), as a newly discovered class of triterpenoid GT from G. uralensis. GuCSyGT expressed in yeast, which could transfer the sugar glucuronic acid to the C3 position of glycyrrhetinic acid and soyasapogenol B, which are the sapogenins of glycyrrhizin and soyasaponin I, respectively. This suggested that GuCSyGT is involved in the biosynthesis of glycyrrhizin and soyasaponin I. However, the in planta role of GuCSyGT in saponin biosynthesis remains unclear. In this study, we generated GuCSyGT-disrupted licorice hairy roots using CRISPR–Cas9-mediated genome editing and analyzed the saponin content. This revealed that soyasaponin I was completely absent in GuCSyGT-disrupted lines, demonstrating the in planta role of GuCSyGT in saponin biosynthesis.