Benzofurans from Eupatorium chinense enhance insulin-stimulated glucose uptake in C2C12 myotubes and suppress inflammatory response in RAW264.7 macrophages.

Fourteen acetylbenzofuran derivatives, including three undescribed carbon skeletons with a newly formed hexane or benzene ring on the other side of the benzofuran ring, (±)-eupatonin A (1), (±)-eupatonin B (2), and eupatonin C (3), two new benzofurans (-)-12ß-hydroxygynunone (4) and (+)-12-hydroxyl-13-noreuparin (5), as well as 9 known ones (6-14), were isolated from 95% ethanol extract of the roots of Eupatorium chinense. Their structures were determined by spectroscopic methods and quantum chemical DFT and TDDFT calculations of the NMR chemical shifts and ECD spectra, which helped in the determination of the relative configurations of 1 and 2 and the absolute configurations of 4 and 5, respectively. 1 and 2 were further identified to be racemic mixtures by chiral HPLC analysis. All compounds were evaluated for insulin-stimulated glucose uptake in differentiated C2C12 myotubes. Compounds 1, 3, 4, 5, 11, 12, and 13 markedly enhanced insulin-mediated glucose uptake. (±)-Eupatonin A (1) activated the IRS-1/Akt/GSK-3ß signaling pathway and enhanced insulin stimulated GLUT4 membrane translocation in C2C12 myotubes. On LPS stimulated RAW264.7 macrophages, several compounds exhibited significant inhibitory effect on NO production with IC50 values ranging from 4.94 to 9.70 µΜ. (±)-Eupatonin A (1) again dose-dependently suppressed LPS-induced NO production and decreased the expression of inducible NO synthase (iNOS), through inhibiting NF-κB activity.

[Chemical constituents from roots of Viburnum setigerum].

This experiment was to study the constituents of the roots of Viburnum setigerum through various column chromatographic techniques. Thirteen compounds were obtained and their structures were identified using chemical and spectroscopic methods as (7αH, 8'αH)-4, 4', 8α-trihydroxy-3, 3', 9-trimethoxy-7, 9'-epoxylignan (1), (7αH, 8'αH)-4, 4', 8α, 9-tetrahydroxy-3, 3'-dimethoxy-7, 9'-epoxylignan (2), alashinol G (3), alashinol F (4), (-)-secoisolariciresinol (5), (7R, 7'R, 8R, 8'S)-3, 3'-dimethoxy-7, 7'-epoxylignane -4, 4', 9, 9'-tetraol (6), (7αH, 8αH, 8'ßH)-4, 4', 7'α, 9-tetrahydroxy-3, 3'-dimethoxy-7, 9'-epoxylignan (7), loganin (8), dihydroquercetin (9), protocatechuic acid (10), 4-hydroxy-3-methoxy-benzoic acid (11), adoxoside (12), and catechin (13). Compound 1 was a new compound. Compounds 3-7 and 11 were reported from the genus Viburnum for the first time. All compounds were separated from this plant for the first time.

Terpenoids from Curcuma wenyujin increased glucose consumption on HepG2 cells.

Thirty four terpenoids, including two new cadinane-type sesquiterpenoids containing conjugated aromatic-ketone moieties, curcujinone A (1) and curcujinone B (2), were isolated from 95% ethanol extract of the root tubers of Curcuma wenyujin. Their structures were determined by spectroscopic methods, especially 2D NMR and HRMS techniques. The relative and absolute configurations of 1 and 2 were identified by quantum chemical DFT and TDDFT calculations of the 13C NMR chemical shifts, ECD spectra, and specific optical rotations. All compounds and extracts were evaluated for their anti-diabetic activities with a glucose consumption model on HepG2 Cells. The petroleum fraction CWP (10µg/mL) and compounds curcumenol (4), 7α,11α-epoxy-5ß-hydroxy-9-guaiaen-8-one (5), curdione (17), (1S, 4S, 5S 10S)-germacrone (18), zederone (20), a mixture of curcumanolide A (25) and curcumanolide B (26), gajutsulactone B (27), and wenyujinin C (30) showed promising activities with over 45% increasing of glucose consumption at 10µM.

Rapid screening of different types of antitumor compound groups from traditional chinese medicine by hollow fiber cell fishing with high performance liquid chromatography.

A novel hollow fiber cell fishing with high performance liquid chromatography (HFCF-HPLC) was extended and used to screen flavonoid and anthraquinone active compound groups simultaneously from traditional Chinese medicines (TCMs). In this study, three cells (MCF-7, SGC7901, and MADB-106) were seeded on the inner wall of the hollow fiber employed to screen bioactive components from TCM water decoction. The variables influencing HFCFHPLC, such as cell seeding time, screening stirring rate and time, and active compound concentration, were investigated and optimized. The surface property of the hollow fiber seeded with cells, the cell survival rate under different conditions, the nonspecific binding between active centers in the fiber and the target compounds, and the repeatability and recovery of HFCF-HPLC were analyzed and validated. Certain structures of the compounds fished by HFCF-HPLC were identified after comparing the retention times of the reference substances. To verify preliminarily the binding site between the bioactive components and cells, we separated the cell membrane and cell organelle from live MCF-7 cells. We then employed the cell membrane, cell organelle, and the whole cells to screen simultaneously the active compounds. The cell fishing factor of the active compound was calculated and discussed as the index of cell-drug binding ability in HFCFHPLC. Tamoxifen as a positive control and indomethacin as a negative control were screened by HFCF-HPLC to verify the method. The results indicate that HFCF-HPLC is an effective and reliable method for the screening and analysis of bioactive components. Moreover, this method can be applied to predict bioactive candidates in TCMs.

A new ionic liquid-water-organic solvent three phase microextraction for simultaneous preconcentration flavonoids and anthraquinones from traditional Chinese prescription.

A novel technique, ionic liquid-water-organic solvent three phase microextraction (ILWOS-3p-ME) was developed and introduced for simultaneous preconcentration and determination of flavonoids and anthraquinones in Chinese herbal formula and its preparations. This technique was performed in one step by using a syringe. High performance liquid chromatography with an UV-detector (HPLC/UV) was subsequently conducted. Two solvents with different densities (organic solvent and ionic liquid with densities less than and higher than water, respectively) were separately placed in a syringe, which was used as an extraction device. A cloudy emulsion was formed by manually shaking the syringe. The mixture was allowed to stand for several minutes; afterward, the emulsion readily separated into three phases: an upper organic solvent extraction phase; a middle aqueous sample phase; and a lower ionic liquid extraction phase. Both the upper and lower layers were transferred to a small Eppendorf (EP) tube. Conducting ILWOS-3P-ME with HPLC/UV, we simultaneously determined the bioactive components of flavonoids and anthraquinones in traditional Chinese medicine. ILWOS-3P-ME is a simple, rapid, practical, and effective method to extract and preconcentrate of different types of trace bioactive components from traditional Chinese medicine simultaneously.

[Effect of gensenoside Rg3 on apoptosis of Hep-2 and expression of HIF-1alha in human laryngeal cancer cell line under anoxic conditions].

OBJECTIVE: To study the mechanism and effect of gensenoside Rg3 on Hep-2 Cell Line during the normoxia and hypoxia. METHODS: Hep-2 Human Laryngeal Cancer Cell Line was cultured under anoxic conditions, and set the normal control group and positive control group (DDP). MTT was used to observe the growth inhibition rates of Hep-2 Human Laryngeal Cancer Cell by Rg3; The cell cycle and cell apoptosis analysis were detected by FCM. Then the expression of HIF-1alpha and VEGF protein was detected by immunohistochemistry and FCM; The expression of HIF-1alpha and VEGF mRNA were detected by transcription-polymerase chain reaction (RT-PCR). RESULTS: Rg3 could significantly inhibit the growth of Hep-2 cells and arrest the cells in G0/G1 phase during normoxia and hypoxia The mRNA and protein expression of HIF-1alpha were dolon-regulated. CONCLUSION: Rg3 can inhibit Hep-2 cells growth by delaying the progress of cell cycle and inhibit the expression of HIF-1alpha during hypoxia, this may be the mechanism of its anti-tumor effect.