This study aimed to investigate the chemical constituents in the water extract of the whole herb of Hedyotis scandens by silica gel, ODS, and MCI column chromatographies together with preparative high-performance liquid chromatography(HPLC). The structures of isolated constituents were identified by NMR, HR-ESI-MS, etc. Thirteen compounds were isolated and identified as methyl 4-benzoyloxy-3-methoxybenzeneacetate(1), 4-benzoyloxy-3-methoxybenzeneacetic acid(2), 3-(4-hydroxy-3-methoxyphenyl)-propanoic acid(3), salicylic acid(4), 3-hydroxy-4-methoxypyridine(5), syringic acid(6), hydroxycinnamic acid(7),(R)-6-methyl-4,6-bis(4-methylpent-3-enyl)cyclohexa-1,3-dienecarbaldehyde(8), 1,2-bis(4-hydroxy-3-methoxyphenyl)-1,3-propanediol(9), 1H-indole-3-carboxaldehyde(10), isoscopoletin(11), syringaresinol(12), and pinoresinol(13). Among them, compounds 1 and 2 were new phenolic acid compounds, compounds 3-5, 8-11, and 13 were isolated from this genus for the first time, and compounds 6, 7, and 12 were obtained from H. scandens for the first time. The activity test showed that compounds 1 and 10 had a certain inhibitory effect on Mycobacterium smegmatis, with MIC_(50) values of 58.5 and 33.3 μg·mL~(-1), respectively.
Hedyotis/chemistry* ; Drugs, Chinese Herbal/chemistry* ; Magnetic Resonance Spectroscopy ; Salicylic Acid

Hedyotis hedyotidea has been traditionally used for the treatment of arthritis, cold, cough, gastro-enteritis, headstroke, etc. But few studies have screened the active compounds from extracts of H. hedyotidea. In this study, the structure of the chemical constituents from stems of H. hedyotidea were determined and the immunosuppressive activity of the compounds was evaluated. The compounds were separated and purified with silica gel, gel column chromatographies and preparative HPLC, and their structures were identified by spectral methods such as MS and NMR. Eleven compounds were obtained and identified as(6S,9S) -vomifoliol (1), betulonic acid (2), betulinic acid (3), betulin(4), 3-epi-betulinic acid (5), ursolic acid (6), β-sitosterol (7), stigmast-4-en-3-one (8), 7β-hydroxysitosterol (9), (3β,7β) -7-methoxystigmast-5-en-3-ol (10) and morindacin (11). This is the first report of compounds 1, 2, 4, 8, 9, 10 and 11 from H. hedyotidea. Compounds 1, 2 and 8-11 were firstly isolated from the genus Hedyotis, and compounds 9 and 10 were isolated from the family Rubiaceae for the first time. The immunosuppressive activity of these compounds was tested using the lymphocyte transsormationtest. Compounds 4, 6 and 9 showed significant immunosuppressive activity.
Animals ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Hedyotis ; chemistry ; Immunosuppressive Agents ; chemistry ; isolation & purification ; pharmacology ; Lymphocytes ; drug effects ; immunology ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred C57BL ; Molecular Structure ; Plant Stems ; chemistry

OBJECTIVE: To explore the proliferation inhibition and apoptosis of polysaccharides extracts from polysaccharides extracts from Hedyotic diffusa (PEHD) on Human Nasopharyngeal Carcinoma (NPC)cell line CNE2 cells in vitro. METHOD: CNE2 cells treated with various concentrations of PEHD were detected by MTT assay at 24 h, 48 h, and 72 h. The apoptotic cells were analyzed by flow cytometry with Annexin V/PI staining. The expression levels of Bax, Bcl-2 and caspase-3 protein were examined by Western blotting method. RESULT: The growth of CNE2 cells were suppressed after treatment with PEHD (P < 0.05), MTT assay showed that the highest cell inhibition rate reached to 76.5%, the inhibition in the doses from 2 to 6 mg/ml showed dose-and-time-dependent. The percent of apoptosis in 4 and 6 mg/ml PEHD treatment groups for 48 h were 31.32%, 46.28%, respectively, and significantly higher than that in control groups, 4.86% (P < 0.01). After the cells being treated with PEHD for 48 h, the expression of Bax and caspase-3 protein increased, and the expression of Bcl-2 protein decreased gradually. CONCLUSION PEHD could inhibited the growth of CNE2 cells and was dose-and-time-dependent, the mechanism may involve induction of cell apoptosis, which was associated with the activation of Bax and caspase-3 protein and the down-regulation of Bcl-2 protein expression.
Apoptosis ; Carcinoma ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Down-Regulation ; Hedyotis ; chemistry ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; pathology ; Plant Extracts ; pharmacology ; Polysaccharides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the proliferation inhibition and apoptosis effects of polysaccharides extracts from Hedyotis diffusa (PEHD) on multiple myeloma (MM) cell line RPMI 8226 cells in vitro, so as to provide experimental theory for the clinical application in the treatment of MM.

METHODSMTT assay was used to examine the effects of PEHD on cell growth. The apoptotic cells were analyzed by flow cytometry with AnnexinⅤ/PI staining. Hoechst staining was used to observe the morphological changes of RPMI 8226 cell apoptosis. The expression levels of caspase-3,-8,-9, PARP, nucleoprotein NF-κB protein and other channel protein were assayed by Western blotting method.

RESULTSThe growth of RPMI 8226 cells were suppressed after treatment with PEHD, the highest inhibition rate reached to 92.3%, the results in the doses from 1 to 4 mg/ml showed a dose-and-time-dependent manner. The proportion of apoptotic cells in 1, 2 and 3 mg/ml PEHD treatment groups for 24 h were 22.52%, 62.31% and 69.94%, respectively, and significantly higher than that of control 8.93%. After treated with PEHD, apoptotic body appeared in RPMI 8226 cells nucleus and the number of apoptotic body increased in a dose-dependent manner. With the increasing of PEHD concentration, the expression of caspase-8,-9,-3 and PARP protein increased. The expression of Mcl-1, Bcl-xl, Bid and Bim protein decreased gradually, but the expression of Bax, Bak and Bad protein increased, and the expression of p-AKT protein (60 kDa) and NF-κB obviously decreased.

CONCLUSIONPEHD could inhibited the growth of RPMI 8226 cells and displayed a dose-and-time-dependent manner, its mechanism may involve cell apoptosis induction, which was associated with the activation of caspase-8, caspase-9, and caspase-3 protein and the down-regulation of p-AKT and NF-κB protein expression.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Hedyotis ; chemistry ; Humans ; Multiple Myeloma ; metabolism ; pathology ; NF-kappa B ; metabolism ; Polysaccharides ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism

OBJECTIVETo analyze p-coumaril acid, trans-6-O-p-coumaroyl scandoside methyl ester and its metabolites in rat plasma after intragastric administration of exocarpium Diffusa effective extracts.

METHODRat blood samples were collected 1.0 h after oral administration of 0.4 g x kg(-1) exocarpium Diffusa effective extracts, and then were analyzed by using HPLC-Q-TOF-MS.

RESULTIn rat plasma, only one metabolite was detected. The structure was identified by reference substance to be p-coumaril acid.

CONCLUSIONThe methyl glucoside metabolite in rat from the main component parts of trans-6-O-p-coumaroyl scandoside methyl ester droppings of vines in Diffusa effective extracts is p-coumaril acid. This experiment provides a theortical basis for studying chemical compositions and pharmacodynamic action of Diffusa.

Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; metabolism ; Hedyotis ; chemistry ; Male ; Mass Spectrometry ; methods ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the effects of three Chinese herbal antidotes, i.e. Herba Artemisiae annuae (A), Herba Hedyotis diffusae (H) and Rhizoma Cimicifugae (C), all were ingredients of Jiedu Quyu Ziyin Recipe, for adjusting the regulated on activation normal T cell expressed and secreated (RANTES), gene expression in serum and renal tissue of MRL/lpr mice.

METHODSFifty-four MRL/lpr mice were randomized into 9 groups, with 6 in each, and intragastrically infused with A, H, C, A+H, H+C, A+C, A+H+C (all in dosage-form of decoction), prednisone suspension and physiological saline, respectively for 12 weeks. RANTES expression in serum and renal tissue of animals were detected with ELISA and RT-PCR at the end of the study.

RESULTSLevels of RANTES expression was significantly reduced in the prednisone treated group after treatment. Excepting no significant change being observed in the groups treated with A and C, the changes in the other groups were all milder than those in the group treated with A+H+C.

CONCLUSIONChinese herbal antidotes A, H and C in combination can significantly inhibit the RANTES expression in serum and renal tissue of MRL/lpr mice.

Animals ; Artemisia annua ; chemistry ; Chemokine CCL5 ; blood ; metabolism ; Cimicifuga ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hedyotis ; chemistry ; Kidney ; metabolism ; Mice ; Mice, Inbred MRL lpr ; T-Lymphocytes ; immunology ; metabolism

OBJECTIVETo investigate the chemical constituents from Hedyotis diffusa.

METHODThe compounds were isolated and purified by various chromatographic techniques and identified by their physicochemical properties and spectral data.

RESULTEight compounds had been reported in last paper, and this time eight more compounds were isolated and identified as 6-hydroxystigmasta-4,22-dien-3-one (1), 3-hydroxystigmasta-4,22-dien-7-one (2), 2-hydroxy-3-methylanthraquinone (3), 2,6-dihydroxy-3-methyl-4-methoxyanthraquinone (4), iso-scutellarein (5), isoetin (6), aesculetin (7), gypsogenic acid (8).

CONCLUSIONCompounds 1-3, 5-8 were obtained from the genus Hedyotis for the first time.

Hedyotis ; chemistry ; Organic Chemicals ; analysis ; isolation & purification

OBJECTIVETo develop an HPLC method for determination of scandoside methyl ester in Hedyotis chrysotricha.

METHODThe determination was carried out on an HC-C18 column elnted with acetonitrile-water (7:93) as mobile phase, and the detection wavelength was set at 238 nm.

RESULTThere is a good linearity in the range 22.08-552 mg L(-1) of scandoside methyl ester and the average recovery is 97.7% (n = 6), RSD 0.72%.

CONCLUSIONThis method is convenient, quick, and is applicable to the quality control of H. chrysotricha.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Esters ; analysis ; Hedyotis ; chemistry

OBJECTIVEA new method for simultaneous determination of four methyl-3, 4-dihydroxybenzoate ( I ), P-coumaric acid (II), ferulaic acid (III) and E-6-O-P-coumaroyl scandoside methyl ester (IV) in Hedyotis diffusa oral solution by reversed-phase HPLC was developed.

METHODThe separation was performed on a Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) with gradient elution. A-acetonitrile, B-methonal-water-glacial acetic acid (5: 95: 0.25), 0-20 min, 1% -16% A; 2042 min, 16% A; 42-46 min, 16%-20%A; 46-65 min, 20% A. The UV detection was set at 265 nm; flow rate 1.0 mL x min(-1).

RESULTThere was good linearity between the peak area and concentration at the ranges of 2.1-105 (r = 0.999 8), 3.5-175 (r = 0.999 8), 1.72-86 (r = 0.999 9), 4.0-200 mg x L(-1) (r = 1.000 0) for I, II, III and IV respectively. The average recoveries of I, II, III and IV were 99.9%, 97.9%, 98.6% and 98.1%.

CONCLUSIONThe method is rapid, simple and accurate, and it can be used for the evaluation of H. diffusa oral solution.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Hedyotis ; chemistry ; Reproducibility of Results

OBJECTIVETo investigate the chemical constituents from Hedyotis diffusa.

METHODThe compounds were isolated and purified by various chromatographic techniques and identified by their physicochemical properties and spectral data.

RESULTEight compounds were isolated and identified as octadecyl (E)-p-coumarate (1), p-E-methoxy-cinnamic acid (2), ferulic acid (3), scopoletin (4), succinic acid (5), aurantiamide acetate (6), rubiadin (7), robustaquinone D (8).

CONCLUSIONCompounds 1-8 were obtained from genus Hedyotis for the first time.

Anthraquinones ; chemistry ; isolation & purification ; Coumaric Acids ; chemistry ; isolation & purification ; Dipeptides ; chemistry ; isolation & purification ; Hedyotis ; chemistry ; Plant Extracts ; chemistry ; isolation & purification ; Scopoletin ; chemistry ; isolation & purification ; Succinic Acid ; chemistry ; isolation & purification