Hederagenin improves psoriasis skin lesions by inhibiting macrophage mincle-mediated inflammation / 中国临床药理学与治疗学

AIM: To observe the effects and mechanisms of hederagenin (HDG) in improving psoriasis skin lesions and inflammation in mice. METHODS: A mouse model of psoriasis was established by depilation of the back and continuous application of imiquimod for 7 days in C57 mice. After modeling, HDG was administered orally (low dose: 25 mg · kg

Inhibitory effect of hederagenin on the proliferation of bladder cancer cells in vitro and in nude mice through the JNK/p38 MAPK signaling pathway / 中华内分泌外科杂志

Objective:To explore the effect of hederagenin (HE) on the proliferation of bladder cancer cell T24 in vitro and in nude mice.Methods:Human bladder cancer cells T24 were divided into control group and experimental group. The experimental group was cultured with DMEM medium containing 25μg/mL hederogenin, and CCK8 was used to detect cell proliferation ability. Nude mice were divided into a control group and an experimental group and injected with T24 cells. The cells of the experimental group were injected with ivy sapogenin at 30 mg/kg every other day. The protein of T24 cells and tumor mass was extracted to detect the expression of p-JNK/JNK and p-p38/p38.Results:After the bladder cancer cells T24 were treated with hederagenin, the CCK8 results showed that the cell proliferation ability of the experimental group was significantly decreased ( P<0.05) . The expression levels of p-JNK in experimental group and control group were 0.21±0.06 and 0.89±0.15, respectively, and the expression levels of p-p38 were 0.38±0.09 and 1.44±0.26, respectively. The expressions of p-JNK and p-p38 were up-regulated (all P<0.05) . In vivo, it was found that after treatment with ivisaponin, the volume of tumor mass were 1192.07±250.92μm 3 in the subcutaneous tumor experimental group and 2280.50±600.1μm 3 in the control group, and the mass were 0.65±0.29g and 1.62±0.38g, respectively. The mass and volume of the experimental group were decreased (all P<0.05) . We extracted mass proteins, and western blotting results showed that the expression levels of p-JNK in the experimental group and control group were 0.38±0.08 and 1.03±0.19, respectively, and the expression levels of p-p38 were 0.71±0.12 and 1.36±0.25, respectively. The expressions of p-JNK and p-p38 were up-regulated (all P<0.05) . Conclusion:Hederagenin inhibits the proliferation of bladder cancer in vitro and in nude mice through the JNK/p38 MAPK signaling pathway.

Systems pharmacology dissection of action mechanisms for herbs in osteoporosis treatment / 中草药·英文版

Objective: Osteoporosis has become the biggest cause of non-fatal health issue. Currently, the limitations of traditional anti-osteoporosis drugs such as long-term ill-effects and drug resistance, have raised concerns toward complementary and alternative therapies, particularly herbal medicines and their natural active compounds. Thus, this study aimed to provide an integrative analysis of active chemicals, drug targets and interacting pathways of the herbs for osteoporosis treatment. Methods: Here, we introduced a systematic pharmacology model, combining the absorption, distribution, metabolism, and excretion (ADME) screening model, drug targeting and network pharmacology, to probe into the therapeutic mechanisms of herbs in osteoporosis. Results: We obtained 86 natural compounds with favorable pharmacokinetic profiles and their 58 targets from seven osteoporosis-related herbs. Network analysis revealed that they probably synergistically work through multiple mechanisms, such as suppressing inflammatory response, maintaining bone metabolism or improving organism immunity, to benefit patients with osteoporosis. Furthermore, experimental results showed that all the five compounds (calycosin, asperosaponin VI, hederagenin, betulinic acid and luteolin) enhanced osteoblast proliferation and differentiation in vitro, which corroborated the validity of this system pharmacology approach. Notably, gentisin and aureusidin among the identified compounds were first predicted to be associated with osteoporosis. Conclusion: Herbs and their natural compounds, being characterized as the classical combination therapies, might be engaged in multiple mechanisms to coordinately improve the osteoporosis symptoms. This work may contribute to offer novel strategies and clues for the therapy and drug discovery of osteoporosis and other complex diseases.

Research on molecular mechanism of Qiwei Tongbi Oral Liquid in treatment of rheumatoid arthritis via network pharmacology / 中草药

Objective: To investigate the major active components and potential molecular mechanism of Qiwei Tongbi Oral Liquid in treatment of rheumatoid arthritis. Methods: After protein targets related with rheumatoid arthritis (RA) were collected by mining literature, DrugBank, TTD and OMIM database, the potentional protein targets based on molecular docking were projected into KEGG databases to illustrate the molecular mechanism of Qiwei Tongbi Oral Liquid. Then RAW264.7 cells induced by LPS were employed to test the predict results. Results: Data analysis showed that 107 potential components acted on 116 RA related targets and 237 pathways. Herb-compound-target-pathway network analysis showed that triterpenoid, flavonoids, sterols and alkaloids isolated from Qiwei Tongbi Oral Liquid were the main active ingredients. These compounds could interact with a group of targets and pathways that might participate in anti-inflammatory, analgesic, immune regulation, proliferation, apoptosis, migration and invasion of synovial fibroblasts, osteoclast differentiation, migration and bone resorption. Some compounds against anti-inflammatory activity were verified by in vitro. Conclusion: The active components of Qiwei Tongbi Oral Liquid could regulate multiple biological pathways by acting with multiple target proteins, playing a role in reducing inflammation and swelling of joints, preventing and reducing the destruction of joint bones, and promoting the repair of damaged joint bones. The research results not only reveal its molecular mechanism and pharmacodynamic components, but also provide a theoretical basis for the subsequent experimental research of Qiwei Tongbi Oral Liquid.

Identification on triterpenoid saponins of Clematis species by UPLC-LTQ-Orbitrap combined with molecular network / 中草药

Objective: To construct molecular network and analyze rapidly the saponins of six species of Clematis plants. Methods: The mass spectral data, acquired with UHPLC-LTQ Orbitrap MS, were uploaded to the GNPS analysis platform to build molecular network, visualized by Cytoscape software. On the other hand, the triterpenoid saponins from six plants were identified on the basis of the fragmentation regularity of the standard and the reported literature. Results: Twenty-five triterpenoid saponins, including 16 hederagenin saponins and nine oleanolic acid-type saponins, were determined from six kinds of plants. The distribution of the triterpenoid saponins in the six kinds of plants were profiled by the pie chart of each node in molecular network. Twenty compounds were found in at least two species of Clematis. Clematichinenoside A and oleanolic acid 3-O-ribopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranoside were the common constituents in five species. Six saponins were distributed in in single species of Clematis. Conclusion: Compared with traditional phytochemical methods, the molecular network technology of UPLC-LTQ-Orbitrap MS can quickly and visually distinguish different triterpenoid saponins in six kinds of plants. There are similarities and differentiates among the triterpenoid saponins in the six kinds of plants, which provides the basis for the substitution of medicinal materials.

Chemical constituents from Clematis apiifolia / 中草药

Objective: To investigate the chemical constituents from the stem of the Clematis apiifolia. Methods: A variety of chromatographic Methods: were applied in isolation and purification including silica gel, ODS, Sephadex LH-20 gel, and HW-40C and so on. Their structures were identified by the nuclear magnetic resonance (NMR), mass spectrometry and so on. Results: These compounds were isolated and determined as hederagenin (1), α-hederin (2), sapindoside B (3), stigmast-22-en-3β, 6β, 9β-triol (4), pleuchiol (5), stigmasterol-3-O-β-D-glucopyranoside (6), stimasterol (7), siderin (8), shikimic acid (9), ethyl-α-D-glucopyranoside (10), ethyl-β-D-glucopyranoside (11), D-gulonic acid (12), 3,5-dihydroxyl-4-valerolactone (13), 2-formyl-5-hydroxymethylfuran (14), uracil (15), uridine (16), tricosanol (17), triacontanol (18), hexacosanoic acid (19), and monostearin (20). Conclusion: All compounds are obtained from C. apiifolia for the first time including firstly reported compound 3 from Clematis plants and compounds 4, 5, 9-12 from Ranunculaceae family.

Triterpenoids isolated from flower buds of Lonicera macranthoides / 中草药

Objective To study the chemical constituents from the flower buds of Lonicera macranthoides and their antitumor activities. Methods The constituents were separated by chromatography of silica gel, ODS, Sephadex LH20, and semi-pre HPLC. Their structures were elucidated by spectral means. The in vitro cytotoxic activities of the isolated compounds were studied by MTT method. Results Seven compounds were isolated and identified as 3-O-β-D-glucopyranosyl-(1→4)-α-L-arabinopyranosyl- hederagenin 28-O-β-D-glucopyranosyl ester (1), 3-O-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl-oleanolic acid 28-O-α- L-rhamnopyranosyl-(1→4)-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (2), 3-O-α-L-rhamnopyranosyl-(1→2)-α-L- arabinopyranosyl-hederagenin 28-O-β-D-glucopyranosyl ester (3), 3-O-α-L-rhamnnopyranosyl-(1→2)-α-L-arabinopyranosyl- hederagenin 28-O-α-L-rhamnopyransyl-(1→4)-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (4), 3-O-α-L-arabinopyranosyl- hederagenin 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (5), 3-O-β-D-glucopyra- nosyl-(1→4)-α-L-arabinopyranosyl-hederagenin 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside ester (6), and 3-O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl-hederagenin 28-O-α-L-rhamno- pyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (7). Conclusion Compound 1 is a new compound named macranthoidin C, and compounds 2-7 are isolated from L. macranthoides for the first time. Compounds 1, 4, and 5 show cytotoxicities against HeLa cells with IC50 of 54.3, 43.9 and 61.2 μmol/L, respectively.

Chemical constituents of Caulophyllum robustum and its antitumor activities / 中草药

Objective To study the chemical constituents of Caulophyllum robustum and its antitumor activities. Methods The chemical constituents were isolated and purified with silica gel column chromatography, dextran gel Sephadex LH-20, and other chromatographic methods. Their structures were determined by physicochemical and spectral analysis. Furthermore, the cytotoxicity of these chemical components against the NSCLC A549 cell line was measured by CCK-8 method. Results Eleven compounds were isolated and identified as collinsogenin (1), hederagenin (2), eechinocystic acid 3-O-α-L-arabinopyranoside (3), magnolamide (4), (-)-magnocurarine (5), columbamine (6), 4,4’-diphenylmethane-bis (methyl) carbamate (7), (-)-8,8’-dimethoxy-L-O-(β-D- glucopyranosyl) secoisolariciresinol (8), asechipuroside A (9), 2-[4-(3-hydroxy-1-propenyl)-2-methoxyphenoxy]-1,3-propanediol (10), and 1-linoleoylglycerol (11). Conclusion Compounds 1-2 and 4-11 are isolated from Caulophyllum robustum genus for the first time. Compounds 1 and 2 show moderate inhibited effect on the proliferation of NSCLC A549 cell line.

Biosynthesis analysis of hederagenin pathway and its construction in yeast cells / 中国中药杂志

Hederagenin is an effective constituent of many medical plants, such as Clematidis Radix, and has a wide range of applications in anti-tumor, anti-inflammatory, antidepressant, hepatoprotective antibacterial, et al. In order to obtain the efficient production of yeast cells for hederagenin,we successfully cloned and screened out a P450 gene MdMA02 from Malus×domestica which can catalyze oleanolic acid C-23 oxidation with our developed plug and play platform. Its amino acid homology is only 32% as compared to characterized CYP72A68v2. By transforming MdMA02 to the oleanolic acid-producing strain BY-OA, a hederagenin-producing strain was constructed and hederagenin's titer could achieve 101 mg·L⁻¹ using high cell density fermentation, which was 337 times higher than in shake flasks culturing. This study provides a basis for further research on promoting the creation of oleanane-type pentacyclic triterpenoids biosynthetic pathway analysis and relative cell factories construction.

Synthesis and tumor resistance reverse activities ofα-hederagenin-type ring-A fused pyrazine derivatives / 中国药理学与毒理学杂志

α-Hederagenin (H), derived from Hedera nepalensis var.sinensis, is a pentacyclic oleane-type triterpenoid that exhibits clear cytotoxicity to different tumor cell lines.In this study,a series of novel C-28 derivatives of hederagenin (H) were designed and synthesized in attempt to develop potent tumor resistance reverse activities agents. Previous research showed that H6 displayed robust reverse activity for paclitaxel resistance in KBV cells. Importantly, Co-treatment of paclitaxel with H6 significantly reduced the tumor weight to 42%. Pleasingly, H6 enhanced the efficacy of paclitaxel against KBV cancer cell-derived xenograft tumors in nude mice.Mechanism studies had found that H6 activated permeability glycoprotein(P-gp)ATPase,reduced intracellular ATP levels and inhibited efflux of P-gp substrates,thus enhancing the antitumor activity of paclitaxel on KBV cells.Molecular docking analysis of homology P-gp and H6 then conducted using the Surflex-Dock module.H6 showed a high binding affinity docking score with a total score of 5.4148,much higher than that of H(0.1414).The nov-el C-28 derivatives of H was synthesized from H6 via three-step reaction. The reversal activity of all synthesized H derivatives were tested using the MTT assay.The results showed that the derivatives of nitrogen groups at C-28 displayed same even potent activity than parent compound H6.In addition,its underlying mechanism of action and in vivo activity are in explore.

Chemical constituents from Tibetan medicine Swertia chirayita / 中草药

Objective: To investigate the chemical constituents of Swertia chirayita. Methods: Column chromatography, such as silica gel, MCI, Sephadex LH-20 were used to isolate and purify the compounds. Physicochemical properties and spectroscopic methods were used to elucidate their structures. Results: Twelve compounds, including 2 xanthones, 4 triterpenoids, 3 secoiridoids, and 3 other compounds, the chemical constituents were isolated from the ethyl acetate fraction from 85% ethanol extract of S. chirayita, and identified as bellidifolin (1), norbellidifolin (2), oleanolic acid (3), 4-epi-hederagenin (4), 2-epi-corosolic acid (5), ursolic acid (6), amarogentin (7), swerimilegenin I (8), erythrocentaurin (9), pyrocatechol (10), syringic acid (11), and 4-hydroxy-3-methoxybenzoic acid (12). Conclusion: Compounds 4, 5, and 11 are isolated from genus Swertia for the first time, compounds 8 and 9 are found from S. chirayita for the first time.

Saponin constituents from roots and rhizomes of Caulophyllum robustum / 中草药

Objective: To separate the saponins from the roots and rhizomes of Caulophyllum robustum and determine their chemical structures. Methods: The chemical constituents were isolated by repeated silica gel chromatography, medium pressure column chromatography, and semi-preparative liquid chromatography. Their structures were elucidated by the data of NMR and MS. Results: Ten compounds were isolated from the roots and rhizomes of C. robustum and the structures of compounds 1-10 were identified as echinocystic acid-3-O-β-D-glucopyranosyl-(1→2)-α-L-arabinopyranoside (1), 3-O-α-L-arabinopyranosylhederagenin-28-O-β-D- glucopyranosyl-(1→6)-β-D-glucopyranoside (2), HN-saponin H (3), ciwujianosides A1 (4), glycoside L-K1 (5), 3-O-β-D- glucopyranosyl-(1→3)-α-L-arabinopyranosyl-hederagenin-28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-gluco-pyranoside (6), leonticin F (7), 3-O-β-D-glucopyranosyl-(1→3) [β-D-glucopyranosyl-(1→2)] α-L-arabinopyranosyl-echinocystic acid-28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (8), leonticin A (9), and morroniside (10). Conclusion: Compound 10 is a iridoid. Compounds 1 and 10 are firstly isolated from the plants of Caulophyllum Maxim., compouds 2-9 are isolated from this plant for the first time.

Identification of saponins in rhizomes of Anemone davidii by UPLC/Q-TOF-MS/MS / 中草药

Objective: To identify the saponins in the rhizomes of Anemone davidii by ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UFLC/Q-TOF-MS/MS). Methods: The separation was performed on UPLC Welch C18 column (100 mm × 2.1 mm, 1.7 μm), with a mobile phase using 0.1% acetonitrile (A) and water containing 0.1% formic acid (B) for gradient elution. Q-TOF/MS and electrospray ion (ESI) source were applied for the analysis under the negative ion mode, and the running time was 40.25 min. Using target compound screening method, the structures of monitored chemical constituents were identified by retention time, exact relative molecular mass, and cleavage fragments of MS/MS. Results: Fifty-two triterpenoids were separated and identified from the methanol extract of A. davidii, 47 triterpenoids were identified for the first time from A. davidii among which nine pairs of structural isomers were included. Conclusion: UPLC/Q-TOF-MS/MS method can identify the main chemical constituents from A. davidii rapidly and accurately, which develops a new strategy for identification of the chemical constituents in A. davidii.

Saponins, tannins and flavonols found in hydroethanolic extract from Periandra dulcis roots

Periandra dulcis Mart. ex Benth. Fabaceae (Syn.: P. mediterranea (Vell.) Taub.) is native to the northern and middle parts of Brazil. In Brazilian ethnomedicine, their roots are used as anti inflammatory, expectorant, diuretic and laxative. An HPLC-ESI-MS/MS system was employed to provide a rapid method to make a tentative characterization of the compounds found in the hydroethanolic extract from P. dulcis roots. The structures of sixteen compounds found in this hydroethanolic extract were suggested mainly by MS data conjugated with the UVDAD spectra, reference compounds and available mass spectra data in literature. Saponin derivatives of hederagenin and soyasapogenol E, such as hederagenin-3-O-rhamnosyl glucosyl glucuronide, soyasapogenol E-3-O-rhamnosyl glucosyl glucuronide and periandrin isomers were found as the main constituents, with a minor content of flavonols quercetin and myricetin glycosides derivatives and hydrolysable tannins, such as dihexahydroxydiphenoyl galloyl glucoside and trisgalloyl hexahydroxydiphenoyl glucose.To the best of our knowledge, with exception of periandrins found in the roots, nothing has been published about the chemical composition of P. dulcis..

Identification of main chemical constituents in extracts of Akebiae Fructus by HPLC-ESI-MS / 中草药

Objective: To establish an HPLC-ESI-MS method for quickly identifying the chemical constituents in the extracts of Akebiae Fructus. Methods: The main saponin components in the extracts of Akebiae Fructus were detected with the HPLC-ESI-MS in negative ion mode. These components were further analyzed by MS2 and MS3 spectra, and by comparing with the corresponding reference substances and literature data. Results: Seventeen saponins in the extracts of Akebiae Fructus were well separated in one run. Conclusion: The new method is accurate and rapid. It could be used to identify the main chemical constituents in the extracts of Akebiae Fructus and be suitable for the quality control of Akebiae Fructus.

Chemical constituents of Quercus pannosa / 中草药

Objective: To study the chemical constituents of Quercus pannosa. Methods: The compounds were isolated and purified by chromatography on silica gel and RP C18 columns, and HPLC, and their structures were assigned on the basic of spectroscopic data. Results: From the ethyl acetate fraction of 95% ethanol extract from Q. pannosa, eight compounds were isolated. Among them, four were lignans and other four were triterpenoids. They were identified as rel-(7α, 8β)-3-methoxy-4', 7-epoxy-8, 3'-oxyneolignan-4, 9, 8'-triol (1), rel-(7α, 8β)-3-methoxy-4', 7-epoxy-8, 3'-oxyneolignan-4, 9, 9'-triol (2), (+)-syringaresinol (3), (8R, 8'R)-4, 4'-dihydroxy-3, 3'-dimethoxylianane-7, 7'-dione (4), ursolic acid (5), oleanolic acid (6), maslinic acid (7), and hederagenin (8). Conclusion: Compound 1 is a new lignan, named quercus pannosa-triol. The other compounds are isolated from this plant for the first time.

Chemical constituents in Akebiae Fructus based on UFLC-Q-TOF/MS / 中草药

Objective: To identify the chemical constituents in Akebiae Fructus by UFLC-Q-TOF/MS method. Methods: The separation was performed on an Acquity UPLC BEH C18 Column (100 mm × 2.1 mm, 1.7 μm), with a mobile phase using 0.1% formic acid-acetonitrile and water containing 0.1% formic acid (B) for gradient elution. The flow rate was 0.3 mL/min, the temperature of column was 40°C with injection volume of 1 μL. TOF/MS and electrospray ion (ESI) source were applied for the qualitative analysis under the negative ion mode, and the full mass scan range was m/z 100-1500. Results: According to MS principle, twenty-five triterpenoids were identified from the methanol extract of Akebiae Fructus and the chemical structures of other nine unknown compounds were deduced. Conclusion: UFLC-Q-TOF/MS method could identify the main chemical constituents in Akebiae Fructus rapidly and accurately, which lays a foundation for the quality control of Akebiae Fructus.

Chemical constituents in roots of Ilex cornuta / 中草药

Objective: To study the chemical constituents of the roots of Ilex cornuta. Methods: The compounds were isolated and purified by silica gel, Sephadex LH-20, medium pressure column chromatography, and semi-preparative liquid chromatography, and their structures were elucidated by chemical properties and spectroscop analyses. Results: Eight compounds were isolated and their structures were identified to be β-sitosterol (1), lupeol (2), betulonic acid (3), hede-ragenin (4), 3β-acetoxy-28-hydroxyurs-12-ene (5), ursolic acid (6), 19α-hydroxy ursolic acid (7), 3β-acetoxy-ursolic acid (8), 23-hydroxyl-methyl ursolate (9), heptanoic acid (10), β-daucosterol (11). Conclusion: Compounds 4,5,8, and 9 are obtained from this genus for the first time, and compound 3 is obtained from this plant for the first time.