likviriton and isoliquiritigenin

Glycyrrhizae Radix is widely used as herbal medicine and is effective against inflammation, various cancers, and digestive disorders. We aimed to develop a sensitive and simultaneous analytical method for detecting glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin, the four marker components of Glycyrrhizae Radix extract (GRE), in rat plasma using liquid chromatography-tandem mass spectrometry and to apply this analytical method to pharmacokinetic studies. Retention times for glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin were 7.8 min, 4.1 min, 3.1 min, and 2.0 min, respectively, suggesting that the four analytes were well separated without any interfering peaks around the peak elution time. The lower limit of quantitation was 2 ng/mL for glycyrrhizin and 0.2 ng/mL for isoliquiritigenin, liquiritigenin, and liquiritin; the inter- and intra-day accuracy, precision, and stability were less than 15%. Plasma concentrations of glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin were quantified for 24 h after a single oral administration of 1 g/kg GRE to four rats. Among the four components, plasma concentration of glycyrrhizin was the highest and exhibited a long half-life (23.1 ± 15.5 h). Interestingly, plasma concentrations of isoliquiritigenin and liquiritigenin were restored to the initial concentration at 4-10 h after the GRE administration, as evidenced by liquiritin biotransformation into isoliquiritigenin and liquiritigenin, catalyzed by fecal lysate and gut wall enzymes. In conclusion, our analytical method developed for detecting glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin could be successfully applied to investigate their pharmacokinetic properties in rats and would be useful for conducting further studies on the efficacy, toxicity, and biopharmaceutics of GREs and their marker components.

Topics: Administration, Oral; Animals; Chalcones; Chromatography, Liquid; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhizic Acid; Male; Plant Extracts; Quality Control; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Licorice is one of the most frequently used Chinese herbs, mainly containing triterpenoids and flavonoids. Three original plants, Glycyrrhiza glabra L., Glycyrrhiza uralensis Fisch., and Glycyrrhiza inflata Bat., are defined as licorice in Chinese pharmacopeia. In this study, 40 G. uralensis samples (Group A), 60 G. glabra samples (Group B, C and D) and 40 G. inflata samples (Group E and F), were used as plant materials, the genetic diversity of samples were determined by gene sequencing technology and the chemotypic diversity were detected by HPLC. The chemotypic diversity analysis showed that contents of triterpenoids in G. glabra (isoglycyrrhizin: 2.483±0.0671 mg∙g

Topics: Chalcones; Chromatography, High Pressure Liquid; DNA, Chloroplast; DNA, Intergenic; Flavanones; Flavonoids; Genetic Variation; Glucosides; Glycyrrhiza; Photosystem II Protein Complex; Plant Proteins; Triterpenes

Licorice is one of the most common herbs in traditional Chinese medicine, and classified as top\ grade in Shen Nong Ben Cao Jing. There are three different original plants of licorice stipulated in Chinese\ Pharmacopeia, Glycyrrhiza uralensis Fisch., Glycyrrhiza glabra L., and Glycyrrhiza inflata Bat. However,\ previous investigation showed that the pharmacodynamic effects of the three licorices were quite different. It is\ very difficult to identify them by the classical identification methods. In order to establish a fast and effective\ identification method, we collected 240 licorice plants from 21 populations of 7 provinces, and amplified their\ ITS and psbA-trnH sequences. ITS sequences with a full length of 616 bp and psbA-trnH sequences with a full\ length of 389 bp were obtained separately. Using DNAMAN to analyze these sequences, 4 variable sites were\ found in ITS sequences and 2 ITS haplotypes were determined, and 3 variable sites were found in psbA-trnH\ sequences and 4 psbA-trnH haplotypes were determined. With the combination analysis of ITS and psbA-trnH\ sequences, the molecular identification method of original licorice was established. Using this method, 40\ samples of licorice slices collected from 4 main herbal material markets in China were identified successfully.\ Furthermore, the contents of 2 triterpenes, 18α-glycyrrhizic acid and 18β-glycyrrhizic acid, and 4 flavonoids,\ liquiritin, isoliquiritin, liquiritigenin, and isoliquiritigenin in these licorice pieces were examined by HPLC and\ the results were analyzed using SPSS 21.0. This study provides a new method in identification of licorice,\ which may serve as a guideline for quality control of licorice slices.

Topics: Chalcone; Chalcones; China; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhiza; Glycyrrhiza uralensis; Glycyrrhizic Acid; Triterpenes

Licorice (Glycyrrhizae radix), the root of Glycyrrhiza uralensis Fisch. (Leguminosae), is mainly used to moderate the characteristics of toxic herbs in Traditional Chinese Medicine, which could be partly interpreted as detoxification. However, the underlying mechanism is still not fully elucidated. Nuclear factor erythroid 2-related factor 2 (Nrf2) plays a key role in the protection against toxic xenobiotics. In our previous research, we have identified that extracts from Glycyrrhiza uralensis induced the expression of Nrf2 nuclear protein and its downstream genes. This research aims to screen the most potent Nrf2 inducer isolated from Glycyrrhiza uralensis and examine its effect on Nrf2 signaling pathway and detoxification system.. Four compounds derived from Glycyrrhiza uralensis (glycyrrhetinic acid, liquiritigenin, isoliquiritigenin and liquiritin) were screened by ARE-luciferase reporter. The most potent ARE-luciferase inducer was chosen to further examine its effect on Nrf2 and detoxification genes in HepG2 cells. The role of Nrf2-dependent mechanism was tested by using Nrf2 knockout mice (Nrf2 KO) and Nrf2 wild-type mice (Nrf2 WT).. ARE-luciferase reporter assay showed these four compounds were all potent Nrf2 inducers, and isoliquiritigenin was the most potent inducer. Isoliquiritigenin significantly up-regulated the expression of Nrf2 and its downstream detoxification genes UDP-glucuronosyltransferase 1A1 (UGT1A1), glutamate cysteine ligase (GCL), multidrug resistance protein 2 (MRP2) and bile salt export pump (BSEP) in vitro and in vivo. Additionally, isoliquiritigenin showed Nrf2-dependent transactivation of UGT1A1, GCLC and MRP2.. Isoliquiritigenin, isolated from Glycyrrhiza uralensis, stimulates detoxification system via Nrf2 activation, which could be a potential protective mechanism of licorice.

Topics: Animals; Antioxidant Response Elements; ATP Binding Cassette Transporter, Subfamily B, Member 11; ATP-Binding Cassette Transporters; Chalcones; Female; Flavanones; Glucosides; Glucuronosyltransferase; Glutamate-Cysteine Ligase; Glycyrrhetinic Acid; Glycyrrhiza uralensis; Hep G2 Cells; Humans; Luciferases, Renilla; Mice, Inbred ICR; Mice, Knockout; Multidrug Resistance-Associated Protein 2; Multidrug Resistance-Associated Proteins; NF-E2-Related Factor 2; RNA, Messenger

Liquiritin, isoliquiritin and isoliquirigenin are the active polyphenols present in Glycyrrhiza uralensis which has been used for the treatment of cancer and its complications. The present study was conducted to evaluate the cytotoxicity and antitumor activity of liquiritin, isoliquiritin and isoliquirigenin on human non-small lung cancer cells including apoptosis-induction, inhibition of apoptotic pathways and to explore the underlying mechanism. Lactate dehydrogenase assays, FITC Annexin V staining assay were performed to evaluate cellular cytotoxicity and apoptosis activity. The results showed that pretreatment with these polyphenols induced apoptosis in A549 cells. Liquiritin, isoliquiritin and isoliquirigenin significantly increased cytotoxicity of, and upregulated p53 and p21 and downregulated the apoptotic pathways. Furthermore, it inhibited cell cycle at the G2/M phase. Western blot analysis showed it significantly decreased the protein expression of PCNA, MDM2, p-GSK-3β, p-Akt, p-c-Raf, p-PTEN, caspase-3, pro-caspase-8, pro-caspase-9 and PARP, Bcl-2 in a concentration-dependent manner while the protein expression of p53, p21 and Bax was increased. In addition, Akt pathway was downregulated. These findings suggest that liquiritin, isoliquiritin and isoliquirigenin inhibited the p53-dependent pathway and showed crosstalk between Akt activities. These active polyphenols can be an alternative agent for the treatment of lung cancer.

Topics: Aldehyde Reductase; Apoptosis; Carcinoma, Non-Small-Cell Lung; Caspase 3; Caspase 7; Cell Line, Tumor; Cell Survival; Chalcone; Chalcones; Cyclin-Dependent Kinase Inhibitor p21; Enzyme Activation; Enzyme Inhibitors; Flavanones; Glucosides; Glycyrrhiza uralensis; Humans; L-Lactate Dehydrogenase; Lung Neoplasms; M Phase Cell Cycle Checkpoints; Plant Extracts; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; Tumor Suppressor Protein p53

Shakuyakukanzoto (SKT) composed of Glycyrrhizae radix (G. radix) and Paeoniae radix (P. radix) has been traditionally used in Japan, Korea and China as an antispasmodic drug for the treatment of skeletal muscle cramps and intestinal cramps.. To evaluate the antispasmodic activity of SKT and its two components, as well as to identify the key constituents of the components which mediate this effect in skeletal muscles in vivo.. An experimental cramp model was constructed to evaluate the effects of peripherally-acting muscle relaxants on electrically-induced cramps under physiological conditions. This was accomplished by surgically isolating the motor supply to the gastrocnemius muscle in an anesthetized rat and delivering electrical stimuli to an isolated tibial nerve to induce tetanic contractions. We first tested dantrolene, a well-known peripherally-acting relaxant, to determine the sensitivity and reliability of our experimental model. We then evaluated the effects of SKT, P. radix, G. radix, and the eight active constituents of G. radix against tetanic contractions.. We found that dantrolene (10 and 30 mg/kg, i.d.) rapidly and significantly inhibited tetanic contractions (P<0.01) irrespective of dose. SKT (0.5, 1.0, and 2.0 g/kg, i.d.) and G. radix (0.5 and 1.0 g/kg, i.d.) also significantly inhibited tetanic contractions (P<0.01) but in a dose-dependent manner owing to the actions of six of the eight active constituents in G. radix (liquiritin apioside, liquiritigenin, isoliquiritin apioside, isoliquiritigenin, glycycoumarin, and glycyrrhetinic acid, 20 μmol/kg, i.v.). These constituents, which include flavonoids, a triterpenoid, and a courmarin derivative, demonstrated temporal variations in their inhibitory activity. In contrast, P. radix (0.5 and 1.0 g/kg, i.d.) did not show a statistically significant antispasmodic effect in our study; however, we previously found that it had a significant antinociceptive effect.. Our findings show that SKT inhibits tetanic contractions in vivo and that G. radix is the main antispasmodic component due to the actions of its active constituents, thus supporting the traditional use of SKT. We further propose that SKT containing the antispasmodic G. radix and antinociceptive P. radix is a pharmaceutically elegant option for muscle cramps as treatment requires a two-pronged approach, i.e., inhibition of hyperexcitable skeletal tissues and modulation of the pain accompanying cramps.

Topics: Animals; Animals, Outbred Strains; Chalcone; Chalcones; Coumarins; Dantrolene; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Combinations; Drugs, Chinese Herbal; Electric Stimulation; Flavanones; Glucosides; Glycyrrhetinic Acid; Glycyrrhiza; Male; Mice; Muscle Contraction; Muscle Cramp; Muscle, Skeletal; Paeonia; Parasympatholytics; Phytotherapy; Plant Roots; Rats; Rats, Wistar; Rotarod Performance Test; Tibial Nerve

An ionic liquid-based ultrasonic-assisted extraction (ILUAE) method had been used for the effective extraction of isoliquiritigenin (IQ), liquiritin (LQ) and glycyrrhizic acid (GA) from licorice. The ionic liquids with different cations and anions were investigated in this work and 0.5 M 1-butyl-3-methylimidazolium bromide solution was selected as solvent. In addition, the technical parameters including soaking time, solid-liquid ratio, ultrasonic power and time were optimized. Compared with the conventional solvent extraction, the proposed approach exhibited higher efficiency, which indicated the ILUAE was an efficient, rapid and simple sample preparation technique. There was no degradation of the target analytes had been observed at the optimum conditions which was evidenced by the stability studies performed with standard of IQ, LQ and GA. The proposed method also showed high reproducibility and was environmental friendly.

Topics: Chalcones; Chemical Fractionation; Chromatography, High Pressure Liquid; Flavanones; Glucosides; Glycyrrhiza; Glycyrrhizic Acid; Ionic Liquids; Plant Extracts; Ultrasonics

A high performance liquid chromatography coupled with photodiode-array detection method was developed for simultaneous determination of glycyrrhizin, paeoniflorin, benzoic acid, quercetin, ferulic acid, formononetin, liquiritin and isoliquiritigenin in the Chinese proprietary medicine "Xiao Yao Wan" (XYW). The analysis was performed by reverse phase gradient elution, using an aqueous mobile phase (containing 0.1% phosphoric acid) modified by acetonitrile and detection made simultaneously at four wavelengths. The method was validated for accuracy, precision and limits of detection and quantification. Ten batches of XYW obtained from different pharmaceutical companies were analyzed and found to contain different amounts of the eight bioactive markers. This method could be used for quality assessment of this herbal medicine.

Topics: Benzoates; Bridged-Ring Compounds; Chalcones; Chromatography, High Pressure Liquid; Coumaric Acids; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhizic Acid; Isoflavones; Medicine, Chinese Traditional; Molecular Structure; Monoterpenes; Quercetin; Reference Standards; Reproducibility of Results; Sensitivity and Specificity