A tannin compound from Sanguisorba officinalis blocks Wnt/ß-catenin signaling pathway and induces apoptosis of colorectal cancer cells.

BACKGROUND: Sanguisorba officinalis, a popular Chinese herb, called DiYu, has been shown to inhibit the growth of many human cancer cell lines, including colorectal cancer cells. The aims of this study were to discover the active compound and molecular mechanism of S. officinalis against Wnt/ß-catenin signaling pathway and develop Wnt inhibitors from natural products as anti-colorectal cancer agents. METHODS: 1,4,6-Tri-O-galloyl-ß-d-glucopyranose (TGG) was obtained by the preparative HPLC. The effect of DiYu on proliferation of NIH3T3 and HT29 was detected by MTT assay. Luciferase reporter assay was applied to investigate the activity of Wnt/ß-catenin signaling in NIH3T3. The expression levels of mRNA and protein were detected by RT-PCR and western blot. Immunofluorescence assay was used to measure the level of ß-catenin in cytoplasm and nucleus. Transcriptomic profiling study was performed to investigate the molecular mechanism of DiYu on the Wnt/ß-catenin signaling pathway. RESULTS: TGG significantly inhibited the Wnt/ß-catenin signaling pathway, down-regulated the expression of ß-catenin and Wnt target genes (Dkk1, c-Myc, FGF20, NKD1, Survivin), up-regulated the levels of cleaved caspase3, cleaved PARP and ratio of Bax/Bcl-2, which may explain the apoptosis of HT29. CONCLUSIONS: Our study enhanced the discovery of the materials and elucidation of mechanisms that account for the anti-Wnt activity of natural inhibitor (DiYu) and identified the potential of TGG to be developed as anti-colorectal cancer drugs.

Chemical constituents of Eleutherococcus sessiliflorus extract and its sedative-hypnotic effect.

The present study was designed to investigate the chemical constituents and bioactivities of the roots of Eleutherococcus sessiliflorus (E. sessiliflorus). The compounds were isolated through various chromatography techniques and elucidated by mass spectrometric (MS), 1D- and 2D-NMR analyses. The sedative-hypnotic effect of E. sessiliflorus ethanol extract and its fractions (acetic ether, n-butanol and water fraction) were also investigated. In the chemical constituent study, one new compound, 3-ethyl-5-hydroxy-3-(hydroxymethyl) pentyl-3-(3,4-dihydroxyphenyl) acrylate and fourteen known compounds were isolated and identified. The chromatographic fingerprint of E. sessiliflorus was established by UPLC-PDA-MS/MS analysis. In bioactivity study, it was found that the water fraction of E. sessiliflorus extract could prolong pentobarbital-induced sleeping time more than that of the other fractions of E. sessiliflorus extract in mice, which provided a basis for future study on sedative-hypnotic effect of the chemical constituents in E. sessiliflorus.

Sanguisorba officinalis L synergistically enhanced 5-fluorouracil cytotoxicity in colorectal cancer cells by promoting a reactive oxygen species-mediated, mitochondria-caspase-dependent apoptotic pathway.

Sanguisorba officinalis L. radix is a widely used herb called DiYu (DY) in China and has an extensive range of bioactivities, including anti-cancer, anti-inflammatory, and anti-oxidative activities. However, there is little evidence to support its anti-cancer effects against colorectal cancer (CRC). The first-line chemotherapeutic agent 5-fluorouracil (5-FU) is used to treat CRC, but its efficiency is hampered by acquired drug resistance. This study found that a water extract of DY exerted anti-proliferative effects against two CRC cell lines (HCT-116 and RKO), and it sensitized CRC cells to 5-FU therapy by activating a reactive oxygen species (ROS)-mediated, mitochondria-caspase-dependent apoptotic pathway. Co-treatment of DY and 5-FU significantly elevated ROS levels, up-regulated Bax/Bcl-2 ratio and triggered mitochondrial dysfunction, followed by a release of cytochrome c and up-regulation of proteins such as cleaved-caspase-9/3 and cleaved-PARP. Additionally, the induction of autophagy may be involved in mediating synergism of DY in HCT-116 cells. Gallic acid (GA), catechinic acid (CA) and ellagic acid (EA) were identified as the potential chief constituents responsible for the synergistic effects of DY. In conclusion, co-treatment of DY, specifically GA, CA and EA, with 5-FU may be a potential alternative therapeutic strategy for CRC by enhancing an intrinsic apoptotic pathway.

Simultaneous Determination of Eight Alkaloids in Rat Plasma by UHPLC-MS/MS after Oral Administration of Coptis deltoidea C. Y. Cheng et Hsiao and Coptis chinensis Franch.

A ultra-high performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method was successfully developed and validated for the identification and determination of eight alkaloids: tetrahydropalmatine (A); palmatine (B); magnoflorine (C); columbamine (D); berberine (E); worenine (F); berberrubine (G) and coptisine (H) in rat plasma, which are the active components in Coptis deltoidea C. Y. cheng et Hsiao (CCY) and Coptis chinensis Franch (CF). The chromatographic separation of analytes was successfully achieved on an Agilent SB-C18 column (1.8 µm, 150 mm × 2.1 mm) using a programme with a mobile phase consisting of acetonitrile and water containing 0.3% acetic acid at a flow rate of 0.25 mL/min. The analytes were detected with a triple quadrupole tandem MS in multiple reaction monitoring (MRM) mode and an electrospray ionization (ESI) source in positive mode. The validated method showed good linearity over a wide concentration range (r² > 0.991), and lower limits of quantification (LLOQ) less than 1.1 ng/mL for all analytes, and matrix effects ranged from 85.2% to 106.8%. The mean extraction recoveries were no less than 86.4%, and the precision and accuracy were within the acceptable limits. All analytes were proven to be stable during sample storage and analysis procedures. The method validation results demonstrated that the proposed method was sensitive, specific, and reliable, which could lay a foundation for the pharmacokinetic study of eight analytes after oral administration of CCY and CF in subsequent studies.

UHPLC-MS/MS Determination, Pharmacokinetic, and Bioavailability Study of Taxifolin in Rat Plasma after Oral Administration of its Nanodispersion.

A rapid and sensitive LC-MS/MS method based on the Triple Quad system has been developed and validated for the determination and pharmacokinetics of taxifolin and its nanodispersion in rat plasma. Taxifolin plasma samples along with butylparaben (internal standard) were pre-treated by liquid-liquid extraction with ethyl acetate, and then separated on a SB-C18 RRHD column (150 mm × 2.1 mm × 1.8 µm) using isocratic elution with a run time of 3.0 min. The mobile phase was acetonitrile-water (90:10, v/v) containing 5 mM ammonium acetate at a flow rate of 0.4 mL/min. Quantification of taxifolin was performed by the electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode with negative atmospheric ionization at m/z 303.0→285.0 for taxifolin and 193.1→92.0 for I.S., respectively. The calibration curve of taxifolin showed good linearity over a concentration range of 5.0-4280 ng/mL with a correlation coefficient of 0.9995. The limit of quantification (LLOQ) was 5.0 ng/mL. Intra-day, inter-day precision and accuracy (percent relative to standard deviation) were all within 8% at three concentration levels. A total recovery of taxifolin and I.S. was beyond 75%. The present LC-MS/MS method was successfully applied to pharmacokinetic studies of taxifolin after intravenous administration of taxifolin, oral administration of its physical mixture and nanodispersion. The absolute bioavailability of taxifolin was calculated as 0.75% for taxifolin nanodispersion and 0.49% for taxifolin, respectively.

Triterpenoids and Flavonoids from the Leaves of Astragalus membranaceus and Their Inhibitory Effects on Nitric Oxide Production.

Four new cycloartane triterpenes, named huangqiyegenins V and VI and huangqiyenins K and L (1-4, resp.), together with nine known triterpenoids, 5-13, and eight flavonoids, 14-21, were isolated from a 70%-EtOH extract of Astragalus membranaceus leaves. The structures of the new compounds were elucidated by detailed spectroscopic analyses, and the compounds were identified as (9ß,11α,16ß,20R,24S)-11,16,25-trihydroxy-20,24-epoxy-9,19-cyclolanostane-3,6-dione (1), (9ß,16ß,24S)-16,24,25-trihydroxy-9,19-cyclolanostane-3,6-dione (2), (3ß,6α,9ß,16ß,20R,24R)-16,25-dihydroxy-3-(ß-D-xylopyranosyloxy)-20,24-epoxy-9,19-cyclolanostan-6-yl acetate (3), and (3ß,6α,9ß,16ß,24E)-26-(ß-D-glucopyranosyloxy)-16-hydroxy-3-(ß-D-xylopyranosyloxy)-9,19-cyclolanost-24-en-6-yl acetate (4). All isolated compounds were evaluated for their inhibitory activities against LPS-induced NO production in RAW264.7 macrophage cells. Compounds 1-3, 14, 15, and 18 exhibited strong inhibition on LPS-induced NO release by macrophages with IC50 values of 14.4-27.1 µM.

[Chemical constituents from Hydrangea macrophylla].

OBJECTIVE: To investigate chemical constituents from Chinese herbal medicine Hydrangea macrophylla. METHOD: The compounds were separated and purified by column chromatography over silica gel, ODS, and preparative HPLC. Their structures were identified by spectral methods including 1H, 13C-NMR and MS. RESULT: Eleven compounds were isolated and identified as zeorin, hopane-6, 22-diol (1), botulin (2), betulinic acid (3), 2-ethyl-3-methyl-maleimide-N-beta-D-glucopyranoside (4), uridine (5), thymidine (6), adenosine (7), nicotinamide (8), methyl pyroglutamate (9), hydrangenol (10) and hydrangenol-4'-O-beta-D-glucopyranoside (11), respectively. CONCLUSION: Compounds 14 and 7-9 were obtained from the genus Hydrangea for the first time.

Monoterpenoids from Acanthopanax sessiliflorus fruits.

Three new acyclic monoterpenoids named (2E)-3,7-dimethylocta-2,6-dienoate-6-O-α-L-arabinopyranosyl-(1→6)-ß-D-glucopyranoside (1), (3Z,6E)-3,7-dimethyl-3,6-octadiene-1,2,8-triol (2) and (6E)-7-methyl-3-methylene-6-octene-1,2,8-triol (3) were isolated from Acanthopanax sessiliflorus fruits, along with three known monoterpenoid compounds. The structures of the new compounds were determined by means of extensive spectroscopic analysis (1D, 2D NMR and HRESIMS) and chemical methods.

Two new cyanogenic glucosides from the leaves of Hydrangea macrophylla.

Chemical investigation of the ethanol extract of the aerial parts of Hydrangea macrophylla collected in the Sichuan Province of China resulted in the isolation of two new cyanogenic glucosides. Their structures were elucidated as [(2R)-2-ß-D-glucopyranosyloxy)-2-(3,4-dimethoxy-phenyl)] acetonitrile (1) and {(2R)-2-[α-D-glucopyranosyl(1-->6)ß-D-glucopyranosyloxy]-2-(3-hydroxy-4-methoxy-phenyl)}acetonitrile (2) on the basis of extensive spectroscopic analysis (1D, 2D NMR and HRESIMS) and chemical studies.