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Isoliquiritigenin (ISL) is an active chalcone compound isolated from licorice. It possesses anti-inflammatory and anti-oxidative activities. In our previous study, we uncovered a great potential of ISL in treatment of type 2 diabetes mellitus (T2DM). Therefore, this study aims to reveal the mechanism underlying the alleviatory effects of ISL on T2DM-induced glycolipid metabolism disorder. High-fat-high-sugar diet (HFD) combined with intraperitoneal injection of streptozotocin (STZ) were used to establish T2DM mice model. All animal experiments were carried out with approval of the Committee of Ethics at Beijing University of Chinese Medicine. HepG2 cells were used in in vitro experiments, and sodium palmitate (SP) was applied to establish insulin resistance (IR) model cells. The effects of ISL on body weight, fasting blood glucose levels, and pathological changes in the livers of mice were examined. Enzyme-linked immune sorbent assay (ELISA) and real-time quantitative PCR (RT-qPCR) were applied to detect the regulatory effects of ISL on key targets involved in glucolipid metabolism. Additionally, molecular docking and analytical dynamics simulation methods were used to analyze the interaction between ISL and key target protein. The results indicate that ISL significantly downregulates the transcriptional levels and inhibits the activities of key enzymes involved in gluconeogenesis, including pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), and fructose-1, 6-bisphosphatase (FBP). It also downregulates the transcriptional and protein levels of hepatocyte nuclear factor 4α (HNF4α) and cAMP response element binding protein (CREB), the two transcriptional factors involved in gluconeogenesis. Thus, ISL inhibits hepatic gluconeogenesis in T2DM mice. In addition, ISL reduces total cholesterol (TC) and triglyceride (TG) levels in the livers of T2DM mice. Moreover, ISL downregulates the mRNA levels of lipogenesis genes and upregulates those of genes involved in fatty acid oxidation, lipid uptake, and lipid export. In conclusion, ISL suppresses hepatic gluconeogenesis, promotes lipolysis, and restrains lipogenesis in T2DM mice, thereby improving the abnormal glycolipid metabolism caused by T2DM.
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OBJECTIVE To study the effects of isoliquiritigenin （ISL） regulating gut microbiota and repairing gut barrier function in model mice with non-alcoholic fatty liver disease （NAFLD）， and to clarify its mechanism for improving NAFLD. METHODS Thirty male C57BL/6J mice were randomly divided into the normal （ultrapure water）， model group （ultrapure water）， ISL group （100 mg/kg）， with 10 mice in each group. Model group and ISL group were fed with high-fat diet for 19 weeks to establish NAFLD model； at the same time， the mice were given relevant medicine/ultrapure water intragastrically. The changes of body weight in mice were recorded， and liver index， white fat index and brown fat index were calculated. The pathological changes of liver tissue and colon tissue as well as lipid accumulation were observed in mice. The levels of total cholesterol （TC）， triglyceride （TG）， aspartate aminotransferase （AST） and alanine aminotransferase （ALT） E-mail：xiangshj3@mail.sysu.edu.cn in serum or liver were measured； the serum levels of interleukin-6 （IL-6）， IL-1β and tumor necrosis factor-α （TNF-α） and the levels of IL-6， IL-1β and TNF-α mRNA expression in liver tissue were detected. Fecal samples underwent 16S rDNA sequencing analysis， and the effects of ISL on gut microbiota structure of mice were investigated. The expressions of gut mucosal barrier-related proteins （Claudin-4， Occludin and ZO-1） were determined in the colon tissue of mice. RESULTS Compared with model group， the body weight， liver index， the levels of TC in liver tissue and serum， the levels of AST and ALT in serum， the levels of IL-6， IL-1β and TNF-α in serum， and the mRNA expression of TNF-α in liver tissue were all decreased significantly in ISL group， while brown fat index was increased significantly. The inflammation and damage of liver tissue were significantly improved， and the NAFLD activity score and the proportion of lipid staining area were significantly reduced （P＜0.05）. ISL could significantly up-regulate the relative abundance of beneficial microbiota （norank_f_Muribaculaceae， Odoribacter， Ruminiclostridium， etc.） and the expressions of intestinal barrier function- related proteins， but could significantly down-regulate the relative abundance of harmful bacteria （Desulfovibrio， norank_f_Lachnospiraceae， unclassified_p_Firmicutes）， and could repair intestinal barrier. CONCLUSIONS ISL could significantly delay the progress of NAFLD， the mechanism of which may be associated with regulating gut microbiota and improving gut barrier function.
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Isoliquiritigenin (ISL) is a flavonoid compound isolated from licorice. It possesses excellent antioxidant and anti-diabetic activities. This study aims to investigate the molecular mechanism underlying the alleviatory effect of ISL on energy metabolism imbalance caused by type 2 diabetes mellitus (T2DM). 8-week-old male C57BL/6J mice were used in in vivo experiments. The high-fat-high-glucose diet combined with intraperitoneal injection of streptozotocin was applied to establish T2DM animal model. All animal experiments were performed in accordance with the Institutional Guidelines of Laboratory Animal Administration issued by the Committee of Ethics at Beijing University of Chinese Medicine. HepG2 cells were used in in vitro experiments. Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to examine the protein and mRNA levels of mitochondrial function-related targets. The levels of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) in HepG2 cells were measured by the flow cytometry. Additionally, the molecular docking of ISL and key target proteins was analyzed. It was found that ISL significantly inhibited the activity of mitochondrial respiratory chain complex I and increased the protein levels of uncoupling protein 2 (UCP2) in the livers of mice and HepG2 cells. It also obviously decreased the ROS levels and increased the MMP levels in cultured HepG2 cells. In addition, ISL promoted mitochondrial biogenesis by activating proliferator-activated receptor gamma co-activator 1α (PGC-1α) and enhanced mitophagy by upregulating Parkin. It also improved mitochondrial fusion by increasing the mRNA and protein levels of mitofusin 2 (MFN2). In conclusion, ISL alleviates energy metabolism imbalance caused by T2DM through suppression of excessive mitochondrial oxidative phosphorylation and promotion of mitochondrial biogenesis, mitophagy, and fusion.
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【Objective】 To investigate the effect of isoliquiritigenin on inflammatory response of vascular endothelial cells and whether the regulatory effect of isoliquiritigenin on inflammation is mediated by histone deacetylase 3 (HDAC3). 【Methods】 Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and treated with LPS, different concentrations of isoliquiritigenin and HDAC3 specific inhibitor, respectively. Real-time PCR and Western blotting were used to detect the mRNA and protein expressions of inflammatory cytokines and HDAC3. Male C57BL/6J mice were randomly divided into vehicle group and isoliquiritigenin treatment group. The vascular inflammation model of C57BL/6J mice was established by ligation of the left carotid arteries. The mRNA expressions of inflammatory cytokines and HDAC3 in the carotid arteries of mice were detected by Real-time PCR. A molecular docking study was performed to investigate the interaction between isoliquiritigenin and HDAC3. 【Results】 Compared with the vehicle group, isoliquiritigenin reduced the mRNA expressions of inflammatory cytokines NLRP3, IL-1β, IL-18, MCP-1 and ICAM-1 and decreased the expression of HDAC3 mRNA and protein in HUVECs stimulated with LPS. In addition, isoliquiritigenin also decreased the mRNA expressions of NLRP3, IL-1β and HDAC3 in carotid arteries of ligated C57BL/6J mice. The docking of isoliquiritigenin in the active site of HDAC3 showed that isoliquiritigenin might act through HDAC3. Furthermore, HDAC3 specific inhibitor RGFP966 further promoted the inhibitory effect of isoliquiritigenin on the expression of inflammatory cytokines in vascular endothelial cells. 【Conclusion】 These results suggest that isoliquiritigenin suppresses the inflammatory response of vascular endothelial cells via HDAC3.
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Abstract As one of the most promising formulations for poorly water-soluble drugs, nanocrystals have been attracting increasing attention in recent years. Isoliquiritigenin (ISL) is a flavonoid with a chalcone structure, and possesses many biological activities. However, its clinical application is significantly limited mainly due to its low oral bioavailability caused by poor hydrophilicity. To address this, ISL nanocrystals were developed in this study to improve its oral bioavailability. Three types of nanocrystals with differing particle size; R1, R2, and R3, were prepared by anti- solvent precipitation or anti-solvent precipitation combined with sonication, which was optimized by single-factor experiments. These nanocrystals were characterized based on their physical properties, in vitro release, and in vivo absorption performance. The mean particle size of R1, R2, and R3 was 555.7, 271.0, and 46.2, respectively. The dissolution ratio of ISL in the nanocrystals was significantly improved, with the quickest rate recorded in R2. Peak concentration and area under the concentration-time curve of R2 after oral administration in rats was 5.83- and 2.72-fold higher than that of the ISL solution, respectively. These findings indicate that the dissolution and absorption of ISL can be significantly enhanced by nanocrystals, and the dissolution behavior and pharmacokinetic properties of nanocrystals is significantly influenced by particle size.
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Objective: To systematically study the main chemical components of Fufang Shangtong Capsule and explore the main mechanism of its effect, and provide some reference for the research of its pharmacodynamic substance. Methods: In this study, UHPLC-Q-Orbitrap HRMS was used to comprehensively analyze the main chemical components of Fufang Shangtong Capsule. The chromatographic column was Waters Acquity UPLC® BEH C18 chromatographic column (50 mm ×2.1 mm, 1.7 μm) and the mobile phase was acetonitrile (A)-0.1% formic acid water (B). According to the MS/MS spectrometry information of compounds, and the comparison with standards or references, the chemical information of drugs can be quickly and accurately identified. On this basis, the network pharmacology method was used to analyze the chemical composition target of the drug, enrich its function, preliminarily select the main effective substances of the drug, and simultaneously explore its mechanism of action. Results: A total of 36 chemicals were identified in this study from Fufang Shangtong Capsule. The target function of enrichment analysis showed that the drug mainly played its therapeutic effect on regulating vascular endothelial, vascular smooth muscle pain, affecting platelet function, promoting energy supply, reducing inflammation and relieving pain, so as to exert its efficacy in promoting blood circulation and removing stasis, invigorating qi and relieving pain. Conclusion: In this study, UHPLC-Q-Orbitrap HRMS combined with network pharmacology was used, wich provided scientific theoretical basis and important reference for the identification of effective ingredients, screening of quality markers and the study of potential mechanism of action of Fufang Shangtong Capsule.
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Objective: To investigate the effects of five kinds of flavonoids (calycosin, formononetin, ononin, isoliquiritigenin, and medicarpin) from Hedysari Radix on promoting osteogenic differentiation of rat bone marrow stromal cells (rBMSCs) and rat calvarial osteoblasts (ROBs). Methods: rBMSCs were isolated according to plastic adherence. ROBs were isolated by enzyme digestion method. The proliferation of rBMSCs and ROBs were detected by MTT assay. ALP activity and calcium content of rBMSCs and ROBs cells were detected by alkaline phosphatase kit and calcium kit. Mineralized nodule formation was detected by alizarin red staining. Results: The five components could promote proliferation, increase ALP activity, increase calcium content, and increase the area and number of calcified nodules of rBMSCs and ROBs (P < 0.05). Among them, calycosin had the best effect on promoting the osteogenic differentiation of rBMSCs, and medicarpin promoted the osteogenic differentiation of ROBs with the best effect, followed by calycosin. Conclusion: Five flavonoids promoted the improvement of osteogenic function, while calycosin has better osteogenic activity on rBMSCs and ROBs and can be used as an excellent osteoinductive factor.
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Objective To study the effects of Chinese herb ingredients with different properties on transporters (URAT1 and OAT4) involved in renal urate reabsorption and serum uric acid level in acute hyperuricemia mice. Methods The OAT4, URAT1- overexpressed monoclonal cell line (MDCK-hOAT4, HEK293-hURAT1) was constructed. The inhibition effect and the half maximal inhibitory concentration (IC50) of different ingredients to transport activity of OAT4 and URAT1 mediating 14C-uric acid were determined. The effects of protocatechuic, liquiritigenin and isoliquiritigenin on serum uric acid levels in acute hyperuricemia mice were studied by the acute hyperuricemia mice induced by potassium oxonate and xanthine. Results The results indicated that nobiletin,liquiritigenin, isoliquiritigenin, licochalcone A with bitter flavor showed strong inhibition to OAT4. The IC50 of nobiletin, liquiritigenin, isoliquiritigenin, and licochalcone A on OAT4 were 0.556 μmol/L, 18.40 μmol/L, 6.831 μmol/L, and 6.825 μmol/L, respectively. Protocatechuic acid and liquiritigenin showed strong inhibition to URAT1 with IC50 of 7.709 μmol/L and 14.54 μmol/L, respectively. Liquiritigenin can significantly reduce the level of serum uric acid of acute hyperuricemia mice, increase the excretion of uric acid, and reduce the level of serum creatinine and blood urea nitrogen. Conclusion Nobiletin, liquiritigenin, isoliquiritigenin and licochalcone A can inhibit the transport activity of OAT4, while protocatechuic acid and liquiritigenin can inhibit the transport activity of URAT1. Liquiritigenin can significantly reduce the level of serum uric acid in acute hyperuricemia mice and protect kidney, the mechanism of which may be associated with the decreasing reabsorption of uric acid by inhibiting the activity of URAT1 and OAT4.
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Objective: To investigate the chemical compounds from the twigs of Morus multicaulis. Methods: The chemical constituents were isolated by various chromatographic methods from 90% ethanol aqueous extract of M. multicaulis and the structures were elucidated by a combined analysis of physicochemical properties, as well as spectroscopic methods including UV, IR, MS ECD and NMR. Results: Six flavonoids were isolated and the structures were identified as moritwigsone A (1), carpachromene (2), isobavachalcone (3), isoliquiritigenin (4), apigenin (5), and quercitrin (6). The absolute configuration of compound 1 was determined by electronic circular dichroism (ECD) calculation. Conclusion: Compound 1 is a new compound, named moritwigsone A, and compounds 2-4 are isolated from this plant for the first time.
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Isoquiritigenin,one of the active constituents in the Chinese herb liquorice,is found to have moderate inhibitory activity against rat monoamine oxidase B(MAO-B,IC5047. 2 μmol·L-1). However,the structure-activity relationship(SAR) remains unclear until now. In an attempt to reveal the SAR of inhibition by isoquiritigenin,and to identify more potent and selective inhibitors of MAOB,a series of 13 derivatives based on the scaffold of isoquiritigenin were prepared,and their purities and structures were confirmed by UPLC,1 H-NMR,13 C-NMR and HRMS. These compounds were then evaluated for their ability to inhibit the enzymatic activity of human MAO-B. The SAR of inhibition was summarized and a potent compound C8 with high inhibitory activity(IC501. 4 μmol·L-1) and selectivity(>57 folds over MAO-A) was identified. Enzyme kinetics studies suggested that C8 acted as a competitive inhibitor. In addition,C8 showed little cytotoxicity to glial cells in vitro,which could be a promising lead compound for further study.
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Animals , Humans , Rats , Drugs, Chinese Herbal , Monoamine Oxidase , Monoamine Oxidase Inhibitors , Plant Extracts , Structure-Activity RelationshipABSTRACT
Objective: To investigate the anticancer effect of isoliquiritigenin (ISL) on human clear cell renal cell carcinoma 786-O cells, and explore its possible molecular mechanism. Method: Thiazolyl blue tetrazolium bromide (MTT) assay was used to detect effect of ISL (0, 10, 25,50, 75, 100 μmol·L-1) on proliferation of 786-O cells. The effect of ISL on migration and invasion of 786-O cells was detected by cell scratch test and Transwell assay. The autophagy was observed under the fluorescence microscope through acridine orange staining and Ad-GFP-LC3 transfection experiment. Western blot was used to detect the expression of autophagy related protein and analyze the changes of phosphatidylinositol-3-kinase (PI3K)/protein kinase B(Akt)/mammalian target of rapamycin (mTOR) signaling pathway to explore the possible mechanism. Result: MTT results showed that ISL could significantly inhibit the proliferation of 786-O cells in a time-dose dependent manner (PPPPPPPPConclusion: ISL can inhibit the proliferation, migration and invasion of clear cell renal carcinoma 786-O cells, and induce autophagy by inhibiting the PI3K/Akt/mTOR signaling pathway.
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Objective To study the chemical constituents of Lianhua Qingwen Capsule (LQC). Methods The compounds were isolated and purified by column chromatography over silica gel and sephadex LH-20, and preparative RP-HPLC. Their structures were elucidated by physicochemical properties and spectral analyses. Results Twenty compounds were isolated and identified as aloe-emodin (1), phillygenol (2), cepharanone B (3), pinoresinol (4), epipinoresinol (5), pinoresinol monomethyl ether (6), piperolactam A (7), (E)-ethyl 3-(4-hydroxyphenyl) acrylate (8), formononetin (9), isoliquiritigenin (10), naringenin (11), kaempferol (12), citreorosein (13), rhein (14), physcion-8-O-β-D-glucopyranoside (15), 4,5-dioxodehydroasimilobine (16), kaempferol-3-O-α- L-ramnopyranosyl-4″-O-E-(4-hydroxy)-cinnamoyl (17), chrysophanol-1-O-β-D-glucopyranoside (18), chrysophanol-8-O-β-D- glucopyranoside (19), and emodin-8- O-β-D-glucopyranoside (20). Conclusion Compounds 2, 3, 5-17, 19, and 20 are isolated from LQC for the first time. This study establishes the foundation for explaining the efficient substance of LQC.
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Objective:To investigate the influence of isoliquiritigenin on the radiosensitivity of glioma stem cells and demonstrate the potential underlying mechanism. Methods:Glioma stem cells were isolated from SHG44 human glioma cells by serum-free medium. Cell proliferation abilities were detected after isoliquiritigenin treatment and radiotherapy by using Cell Counting Kit-8. The formation of glioma stem cell spheres was observed using an inverted microscope. The protein expression levels of Notch1 signal pathway, NF-κB, and caspase-3 were examined by Western blot analysis. Results:Isoliquiritigenin (10μM) inhibited the formation of tumorspheres at 8 Gy radiation (P<0.05). Isoliquiritigenin (20μM) exerted evident growth inhibitory effect on glioma stem cells. Isoliquiritigenin pre-treatment combined with 4 or 8 Gy radiation reduced the cell radioresistance significantly (P<0.05). The protein expression levels of Notch1 in the isoliquiritigenin and DAPT groups were lower than those of the control at 48 h after isoliquiritigenin treatment (P<0.05). The protein expression levels of P-NF-κB began to increase at 6 and 24 h after 4 Gy radiation with isoliquiritigenin pretreatment (P<0.05). Isoliquiritigenin pretreatment combined with 4 Gy radiation increased the protein expression level of cleaved caspase-3 at 24 h after radiation compared with that of the isoliquiritigenin treatment alone (P<0.05). Conclusion:Isoliquiritigenin may downregulate Notch1 signal pathway and affect different aspects of cell stress and death, including NF-κB- and caspase-3-associated processes, thereby promoting the radiosensitivity of glioma stem cells.
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Bone destruction induced by the metastasis of breast cancer cells is a frequent complication that is caused by the interaction between cancer cells and bone cells. Receptor activator of nuclear factor kappa-B ligand (RANKL) and the endogenous soluble RANKL inhibitor, osteoprotegerin (OPG), directly play critical roles in the differentiation, activity, and survival of osteoclasts. In patients with bone metastases, osteoclastic bone resorption promotes the majority of skeletal-related events and propagates bone metastases. Therefore, blocking osteoclast activity and differentiation via RANKL inhibition can be a promising therapeutic approach for cancer-associated bone diseases. We investigated the potential of isoliquiritigenin (ISL), which has anti-proliferative, anti-angiogenic, and anti-invasive effects, as a preventive and therapeutic agent for breast cancer cell-induced bone destruction. ISL at non-toxicity concentrations significantly inhibited the RANKL/OPG ratio by reducing the production of RANKL and restoring OPG production to control levels in hFOB1.19 cells stimulated with conditioned medium (CM) of MDA-MB-231 cells. In addition, ISL reduced the expression of cyclooxygenase-2 in hFOB1.19 cells stimulated by CM of MDA-MB-231 cells. Therefore, ISL may have inhibitory potential on breast cancer-induced bone destruction.
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Humans , Bone Diseases , Bone Resorption , Breast Neoplasms , Breast , Culture Media, Conditioned , Cyclooxygenase 2 , Neoplasm Metastasis , Osteoblasts , Osteoclasts , Osteoprotegerin , RANK LigandABSTRACT
Objective To study the inhibitory mechanism of soliquiritigenin on tyrosinase.Methods The inhibition effect and inhibitory kinetics of tyrosinase induced by oliquiritigenin were investigated.The effect of isoliquiritigenin on A375 melanoma cell were preliminarily indicated.Results The inhibitory effect of isoliquiritigenin on monophenolase and diphenolase of tyrosinase was good, IC50 was (11.22 ±0.92) μM and (48.53 ±4.75) μM, isoliquiritigenin was a competitive inhibitor, the value of inhibition constant ( Ki ) was ( 12.14 ±0.54 ) μM.Isoliquiritigenin could inhibit the prolifevation of A375 cell significantly, and the tyrosinase activity and melanin synthesis decreased.Conclusion Isoliquiritigenin as a tyrosinase inhibitor, plays an important role in the regulation of melanin, which provides the theoretical basis for the clinical anti skin cancer.
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Aim To investigate the effects of isoliquiri-tigenin(ISL)on C6 glioma cell proliferation and differ-entiation.Methods C6 glioma cells’viability and proliferation were respectively measured by SRB test. Colony formation of C6 glioma cells from different groups was assayed.After culturing the cells from each group,giemsa staining was used to observe cell mor-phology.RT-PCR was applied to detect mRNA expres-sion of GFAP.Western blot was applied to detect the expression of GFAP.Results ISL effectively inhibited the viability of C6 glioma cells when compared with the control group in a concentration-dependent manner (P<0.01).The morphological observation under light mi-croscope showed that:in the control group,most of the undifferentiated C6 cells showed long fusiform and po-lygonal shape.Compared to the control group,the C6 cells treated with ISL revealed alteration in morphology such as astrocytes with smaller smooth,round body and much finer longer,tapering processes.The cloning for-mation rate detection revealed that:the colonies in the control group semerged earlier and were larger than those experimental ones,the cloning formation rate was higher,while almost no effective cells colony emerged in ISL treated groups(P <0.01 ).Western blot and RT-PCR analysis showed that GFAP expression in the ex-perimental groups increased(P <0.01).Conclusion ISL may inhibit the proliferation of C6 glioma cells and induce their differentiation.
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Objective To investigate the effects of isoliquiritigenin on the invasive ability of human gastric carcinoma SGC7901 cells, and its molecular mechanisms thereof. Methods The logarithmic phase human gastric carcinoma SGC7901 cells were divided into control group (normal cell culture fluid) and isoliquiritigenin group (isoliquiritigenin solu?ble in cell culture fluid, the concentrations were 10, 25, 50 and 100 μmol/L respectively). Each group had four repeated holes. The proliferation of SGC7901 cells were detected with MTT assay after 24 h, 48 h and 72 h of culture. The experimen?tal drug concentration and action time were researched for the subsequent experiments. The in vitro invasion abilities of SGC7901 cells were assessed with Transwell test. The expression levels of MMP9, Akt and P-Akt were detected by Western blot assay. Results The proliferation of SGC7901 cells were inhibited by 10μmol/L isoliquiritigenin, which can be signifi?cantly inhibited by 25, 50 and 100μmol/L isoliquiritigenin in a concentration-dependent and time-dependent manner. The half inhibitory concentrations (IC50) of 24, 48 and 72 h were 52.48, 44.49 and 32.50μmol/L, respectively. Therefore, the 25, 50 and 100μmol/L isoliquiritigenin were selected as the subsequent experimental drug concentration, and 24 h was used as the action time. Compared with the control group (209.75±9.29), the membrane cell number of 25μmol/L (138.50±10.15), 50μmol/L (89.50 ± 16.56) and 100μmol/L (45.00 ± 8.08) decreased gradually (F=267.948,P<0.05). There was no signifi?cant difference in the expression level of Akt protein between four groups (F=1.492). The expression levels of P-Akt and MMP9 were gradually decreased with the increase of the isoliquirigenin concentration (F=359.219 and 431.324,P<0.05). Conclusion Isoliquiritigenin can obviously inhibit invasion ability of SGC7901 cells, which may be related to the down reg?ulation of the signal transduction pathway protein PI3K/Akt and the down steam protein MMP9.
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Aim To evaluate the mechanism of apopto-sis induced by the isoliquiritigenin in A375 human ma-lignant melanoma cells. Methods Sulforhodamine B ( SRB) method was used to determine the A375 cell viability;acridine orange/ethidium bromide ( AO/EB) and Hoechst 33258 staining were used to observe the morphological changes of apoptotic cells; flow cytome-try was used to detect A375 cell apoptotic rate;DCFH-DA was applied to determine the changes of total intra-cellular ROS in A375 cells;JC-1 method was used to measure the changes of mitochondrial membrane poten-tial;the kits methods were used to determine the con-tent of ATP, lactic acid and glucose in A375 cell which was treated with different concentrations of isoliquiritigenin. Results Isoliquiritigenin could in-hibit A375 cell proliferation in a concentration-depend-ent manner; A375 cells showed obvious apoptosis charateristics after treatment by isoliquiritigenin, and the apoptosis rate increased with increasing concentra-tion of isoliquiritigenin. The level of total intracellular ROS in A375 cells increased obviously after dealing with different concentrations of isoliquiritigenin;in ad-dition, the mitochondrial membrane potential, the lev-els of intracellular ATP,lactic acid and the level of glu-cose uptake all declined. Conclusions These find-ings demonstrate that isoliquiritigenin can induce apop-tosis of A375 cells. The mechanism may be related to elevation of ROS level and reduction of aerobic glycoly-sis level.
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OBJECTIVE: To evaluate hepatoprotection in Nrf2/ARE oxidation/chemical stress defense pathwaycaused by isoliquiritigenin (Iso) based on analysis of bile acids using profile of bile acids. METHODS: High performance liquid chromatography coupled with quadrupole mass spectrometry (HPLC-MS/MS) was applied to determine the contents of all kinds of endogenous bile acids including free bile acids, taurine conjugates and glycine conjugates. By the luciferase reporter gene assay for screening Nrf2-ARE signal targeting molecular experiments in liquorice extract, isoliquiritigenin was capable of markedly activating Nrf2-driven gene expression, therefore it was used to evaluate hepatoprotection in Nrf2/ARE oxidation/chemical stress defense pathway. RESULTS: Based on the analysis using principle components analysis (PCA), partial least square-discriminant analysis (PLS-DA), Nrf2+/+, Nrf2-/- and Iso Nrf2+/+ groups could be distinguished from their Iso Nrf2-/- group, which suggested that the variance of the contents of bile acids could evaluate hepatoprotection caused by Iso. Bile acids of ursodeoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), cholic acid(CA), sodium taurodeoxycholate hydrate (TDCA), deoxycholic acid (DCA), taurocholic acid(TCA) proved to be important corresponds to Iso induced liver protection according to analysis of partial least square-discriminant analysis (PLS-DA) and the statistical analysis showed that there were significant differences among the four groups. It indicated that UDCA, CDCA, CA, TDCA, DCA and TCA could be considered as sensitive biomarkers of Iso induced liver protection. CONCLUSION: This work can provide the base for the further research on the evaluation and mechanism of hepatoprotection caused by liquorice.
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Isoliquiritigenin (ILTG) is a chalcone compound and shows various pharmacological properties, including antioxidant and anti-inflammatory activities. In recent study, we have reported a novel role of ILTG in sleep through a positive allosteric modulation of gamma-aminobutyric acid type A (GABA(A))-benzodiazepine (BZD) receptors. However, the effect of ILTG in GABA(A)R-mediated synaptic response in brain has not been tested yet. Here we report that ILTG significantly prolonged the decay of spontaneous inhibitory postsynaptic currents (sIPSCs) mediated by GABA(A)R in mouse hippocampal CA1 pyramidal neurons without affecting amplitude and frequency of sIPSCs. This enhancement was fully inhibited by flumazenil (FLU), a specific GABA(A)-BZD receptor antagonist. These results suggest a potential role of ILTG as a modulator of GABAergic synaptic transmission.