[Chemical constituents and their α-glucosidase inhibitory activities of seeds of Moringa oleifera].

The chemical constituents of the seeds of Moringa oleifera were isolated and purified by using Sephadex LH-20, Toyo-pearl HW-40F, silica gel, ODS, and MCI column chromatography. The structures of compounds were identified by high-resolution mass spectrometry, ~1H-NMR, ~(13)C-NMR, HMQC, HMBC, and ~1H-~1H COSY, as well as physicochemical properties of compounds and literature data. Twelve compounds were isolated from 30% ethanol fraction of the seeds of M. oleifera and identified as ethyl-4-O-α-L-rhamnosyl-α-L-rhamnoside(1), ethyl-3-O-α-L-rhamnosyl-α-L-rhamnoside(2),(4-hydroxybenzyl)ethyl carbamate(3),(4-aminophenyl)acetic acid(4), ethyl-α-L-rhamnoside(5), methyl-α-L-rhamnoside(6), moringapyranosyl(7), 2-[4-(α-L-rhamnosyl)phenyl]methyl acetate(8), niaziridin(9), 5-hydroxymethyl furfural(10), 4-hydroxybenzeneacetamide(11), and 4-hydroxybenzoic acid(12). Among them, compounds 1 and 2 are two new compounds, compound 3 is a new natural product, and compounds 4-5 were yielded from Moringa plant for the first time. All compounds were evaluated for α-glucosidase inhibitory activity in vitro. Compound 10 showed excellent inhibitory activity with IC_(50) of 210 µg·mL~(-1).

Six New Phenolic Glycosides from the Seeds of Moringa oleifera Lam. and Their α-Glucosidase Inhibitory Activity.

Plant-derived phytochemicals have recently drawn interest in the prevention and treatment of diabetes mellitus (DM). The seeds of Moringa oleifera Lam. are widely used in food and herbal medicine for their health-promoting properties against various diseases, including DM, but many of their effective constituents are still unknown. In this study, 6 new phenolic glycosides, moringaside B-G (1-6), together with 10 known phenolic glycosides (7-16) were isolated from M. oleifera seeds. The structures were elucidated by 1D and 2D NMR spectroscopy and high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) data analysis. The absolute configurations of compounds 2 and 3 were determined by electronic circular dichroism (ECD) calculations. Compounds 2 and 3 especially are combined with a 1,3-dioxocyclopentane moiety at the rhamnose group, which are rarely reported in phenolic glycoside backbones. A biosynthetic pathway of 2 and 3 was assumed. Moreover, all the isolated compounds were evaluated for their inhibitory activities against α-glucosidase. Compounds 4 and 16 exhibited marked activities with IC50 values of 382.8 ± 1.42 and 301.4 ± 6.22 µM, and the acarbose was the positive control with an IC50 value of 324.1 ± 4.99 µM. Compound 16 revealed better activity than acarbose.

Antihyperuricemia and antigouty arthritis effects of Persicaria capitata herba in mice.

BACKGROUND: Hyperuricemia (HUA) is an important risk factor for gout, renal dysfunction and cardiovascular diseases. The whole plant of Persicaria capitata (Buch.-Ham. ex D. Don) H. Gross, namely Persicaria capitata herba, is a well-known ethnic herb with potent therapeutic effects on urinary tract infections and urinary calculus, yet previous reports have only focused on its effect on urinary tract infections. PURPOSE: To evaluate the therapeutic potential of P. capitata herba against gout by investigating its antihyperuricemia and antigouty arthritis effects and possible mechanisms. METHODS: The ethanol extract (EP) and water extract (WP) of P. capitata herba were prepared by extracting dried and ground whole plants of P. capitata with 75% ethanol and water, respectively, followed by removal of solvents and characterization by UHPLC-Q-TOF/MS. The antihyperuricemia and antigouty arthritis effects of the two extracts were evaluated in a potassium oxonate- and hypoxanthine-induced hyperuricemia mouse model and a monosodium urate crystal (MSUC)-induced acute gouty arthritis mouse model, respectively. The mechanisms were investigated by testing their effects on the expression of correlated proteins (by Western blot) and mRNAs (by RT-PCR). RESULTS: UHPLC-HRMS fingerprinting and two chemical markers (i.e., quercetin and quercitrin) determination were used for the characterization of the WP and EP extracts. Both WP and EP extracts showed pronounced antihyperuricemia activities, with a remarkable decline in serum uric acid and a marked increase in urine uric acid in hyperuricemic mice. Unlike the clinical xanthine oxidase (XOD) inhibitor allopurinol, WP and EP did not show any distinct renal toxicities. The underlying antihyperuricemia mechanism involves the inhibition of the activity and expression of XOD and the downregulation of the mRNA and protein expression of glucose transporter 9 (GLUT9) and urate transporter 1 (URAT1). The extracts of P. capitata herba also demonstrated remarkable anti-inflammatory activity in MSUC-induced acute gouty arthritis mice. The mechanism might involve inhibitory effects on the expression of proinflammatory factors. CONCLUSIONS: The extracts of P. capitata herba possessed pronounced antihyperuricemia and antigouty arthritis effects and were, therefore, promising natural medicines for hyperuricemia-related disorders and gouty arthritis. The use of P. capitata herba for the treatment of urinary calculus may be, at least to some degree, related to its potential as an antihyperuricemia and antigouty arthritis drug.

[A new macrocyclic phenolic glycoside from Sorghum vulgare root].

Eleven compounds were isolated and purified from Sorghum vulgare root extract, through column chromatography over silica gel, MCI gel, and preparative HPLC. Their structures were established by MS, 1 D NMR and 2 D NMR data as sorgholide A(1), ß-sitosterol(2), stigmastero(3), daucosterol(4), 4-methoxycinnamic acid(5), taxiphyllin(6), chlorogenic acid(7), p-hydroxybenzaldehyde(8), succini acid(9), trans-p-hydroxycinnamic acid(10), obtusalin(11). Compounds 4,5 and 9-11 were reported from this species for the first time, and compound 1 is the first 24 ring dimeric double lactonol glycoside formed by reverse polymerization of p-hydroxyphenylacetate glucoside, named sorgholide A.

[Chemical constituents from medical and edible plants of Rosa roxburghii].

Rosa roxburghii, a kind of the medical and edible plants belonging to the Rosaceae family, is widely distributed in the southwest districts of China, especially Guizhou province. Now, by reason of the extensive bioactivities, the plant is widely used in the field of food, health product, drug, and so on. In the course of our continuing search for the bioactive constituents, thirteen compounds were isolated from R. roxburghii, and their structures were determined on the basis of physicochemical property, spectroscopic data and comparison with the literatures, as 2-oxo pomolic acid(1), 1ß-hydroxyeuscaphic acid(2), euscaphic acid(3), arjunic acid(4), tormentic acid(5), kaiiichigeside F1(6), rosamultin(7), arjunetin(8), 2ɑ, 3ɑ, 19ɑ-trihydroxy-olean-12-en-28-oic acid 28-O-ß-D-glucopyranoside(9), 2α, 3α, 19α, 24-tetrahydroxyolean-12-en-28-oic-acid 28-O-ß-D-glucopyranosyl ester(10), pyrogallic acid (11), daucosterol(12), and 1, 2-decanediol(13). Compounds 9 and 10 were firstly obtained from Rosaceae family, and compounds 1,4,5,9-11,13 were isolated from this plant for the first time.

Combating against methicillin-resistant Staphylococcus aureus - two fatty acids from Purslane (Portulaca oleracea†L.) exhibit synergistic effects with erythromycin.

OBJECTIVES: The aims of this study were to identify the active ingredients from Portulaca oleracea L. (PO) that could provide synergism with antibiotics against methicillin-resistant Staphylococcus aureus (MRSA) and their possible mechanisms of resistance inhibition. METHODS: High-speed counter-current chromatography (HSCCC) coupled with gas chromatography-mass spectrometry and a panel of laboratory MRSA strains were used for checkerboard and efflux inhibitory assays. KEY FINDINGS: Linoleic and oleic acids were identified from HSCCC fraction 18 of PO with synergistic antibacterial activity when combined with erythromycin against RN4220/pUL5054. Ethidium bromide efflux inhibitory studies revealed that linoleic and oleic acids may interfere the activity of MsrA pump. By comparing among a panel of linoleic and oleic acids analogues, unsaturated fatty acids in salt form with cis configuration and an increase in number of double bonds were found to further increase the antibacterial activity when used alone or in combination with antibiotics. CONCLUSION: This study reported for the first time that two active ingredients, namely linoleic and oleic acids, were identified from PO with synergistic antibacterial activity when combined with erythromycin against MRSA RN4220/pUL5054 and possibly act by inhibiting the efflux pumps of the bacteria cells.

[Efficacy evaluation of acupoints massage on asthenopia of video display terminal under different exposure dose].

OBJECTIVE: To observe the efficacy differences of acupoints massage for asthenopia of video display terminal (VDT) under different exposure dose. METHODS: One hundred and two cases (204 eyes) were divided into a low exposure group and a high exposure group, fifty-one cases in each group. The same intervention of acupoints massage on Cuanzhu (BL 2), Jingming (BL 1), Sizhukong (TE 23), Sibai (ST 2) and Taiyang(EX-HN 5) were given to the two groups, one acupoint for 5 min and once everyday, one month of which made a course. The symptom score, tear film break-up time (BUT) and Schirmer I test(SIT) were observed before and after treatment. RESULTS: (1) The correlation coefficient of cubic curve model of the exposure dose was the biggest with symptom improvement index (P = 0.000), which indicated that the lower VDT exposure index was, the more obvious the symptom improved. The symptom improvement indices of low exposure group and high exposure group, which were (52.31 +/- 16.65)% and (28.93 +/- 13.35)% respectively, were statistical significant difference (P = 0.000). (2) Compared to before treatment, the levels of BUT and SIT in the two groups were both significantly higher (P < 0.05). Compared with the high exposure group, the levels of BUT and SIT in the low exposure group were increased by 0.826 s (P = 0.022) and 1.029 mm (P = 0.033), respectively, after the impact of BUT and SIT was corrected before the research. CONCLUSION: The acupoints massage can improve the symptoms and ocular physiology for patients with VDT asthenpia, and it is more effective for the low exposure cases.

[Anticancer effect of 5-fluorouracil combined with extract of Rosa roxburghii Tratt on human endometrial adenocarcinoma].

OBJECTIVE: To investigate anticancer effects of 5-fluorouracil (5-FU) combined with CL, extract of Rosa roxburghii Tratt on human endometrial adenocarcinoma cell line (JEC). METHODS: JEC cells cultured in vitro in the logarithmic growth phase were seeded in the culture plate and divided into the control group (RPMI 1640), the positive group (10(-4) mol/L 5-FU), the CL groups (at the dose of 0.01, 0.1, 1, 10, and 100 microg/mL), and the CL (0.01, 0.1, 1, 10, and 100 microg/mL) combined with 5-FU groups. Effects of 5-FU combined with CL on JEC cell growth were drawn and measured by MTT and growth curves. Effects of CL combined with 5-FU on the JEC cell differentiation was analyzed by detecting the reduction capability of nitrobenzene thiocyanate (NBT) and lactate dehydrogenase (LDH) contents in the cultured medium. Effects of CL combined with 5-FU on the JEC cell apoptosis and cell proliferation cycle were detected by acridine orange (AO)/ethidium bromide (EB) fluorescent staining and flow cytometry (FCM). RESULTS: The proliferation inhibitory effect of CL combined with 5-FU on JEC cells was enhanced when compared with that of CL or 5-FU alone (P<0.05). The percentages of NBT positive JEC cells and apoptotic JEC cells increased in the 5-FU combined with CL groups when compared with 5-FU group or the CL group alone (P<0.05). The LDH concentration of the JEC cell culture supernate decreased in 5-FU combined with CL groups (P<0.05). Furthermore, the percentage of G0-G1 phase JEC cells treated by 5-FU combined with CL was higher than that of 5-FU or CL alone (P<0.05). CONCLUSION: CL could enhance anticancer effects of 5-FU. Its mechanisms might be correlated with reinforcing the cytotoxicity of 5-FU, inducing cell differentiation and apoptosis, and inhibiting cell proliferation and division.

[Studies on the chemical constituents of Laportea bulbifera].

OBJECTIVE: To study the chemical constituents of Laportea bulbifera. METHODS: 70% EtOH was used for constituent extraction, silica gel column chromatography for constituent seperation, physical and chemical properties together with spectroscopic methods for chemical structure identification. RESULTS: six compounds were obtained from root of L. bulbifera. Their chemical structures were elucidated as p3-sitosterol(1) , P-daucosterol (2), 2,2'-oxy-bis(1-phenylethanol (3), 1-(2-phenylcarbonyloxy acetyl) benzene (4) , methyl linoleate(5),1,4-diphenyl-1,4-butanedione(6). CONCLUSION: All compounds are isolated from L. bulbifera and among compounds 3 and 4 are reported as natural products for the first time.

[Study on the alkaloids in Stephania hernandifolia].

OBJECTIVE: To study the alkaloids in Stephania hernandifolia. METHODS: The dried herbs of S. hernandifolia were extracted with 95% ethanol. After removal of the solvent, the residue was first partitioned between acid water and petroleum ether, then the aqueous layer was basified and extracted with chloroform to obtain crude alkaloids. Column chromatography on silica gel, Rp-18, MCI CHP 20P, Sephadex LH-20 were applied for the isolation and purification of the crude alkaloids fraction. The structures were elucidated by their physicochemical properties and spectral data. RESULTS: Six alkaloids were obtained and identified as Cepharamine (I), l-tetrahadropalmatine (II), Plmatine (III), Stephamiersine (IV), Telitoxine (V), Daurioxoisoporphine D (VI). CONCLUSIONS: Compounds I - VI are isolated from this plant for the first time, and compounds V, VI are isolated from the plants of Stephania for the first time.

Alkaloids from Daphniphyllum oldhami.

Four new Daphniphyllum alkaloids, daphnioldhanines H-K ( 1- 4), along with 34 known alkaloids, were isolated from Daphniphyllum oldhami. The known alkaloid dehydrodaphnigraciline ( 5) is now reported as a natural product. Their structures were elucidated by spectroscopic methods, especially 2D NMR techniques. The effects against platelet aggregation of compounds 1, 3, and 5 were evaluated, and 3 showed stronger activity against platelet aggregation induced by PAF. This is the first report of quinolizidine alkaloids from the genus Daphniphyllum.

[Anticancer effect of CL extract of Rosa roxburghii].

OBJECTIVE: To investigate anticancer effects and potential mechanisms of CL, extract of Rosa roxburghii. METHOD: In vitro anticancer effect was observed in Ehrlich's ascites carcinoma (EAC) mice model. Cell toxicity of CL on human endometrial adenocarcinoma cell line JEC (JEC) cells was measured by MTT reduction test and growth curves drawing by trypan blue dye exclusion method. Lactate dehydrogenase (LDH) concentration of cultured medium was detected by auto-biochemistry-meter. Cell differentiation was showed by detection of NBT reduction ability. Apoptosis was showed by AO/EB fluorescent staining and flow cytometer detection. Cell proliferation cycle was detected by flow cytometer. RESULT: Comparing with the negative group, life span of EAC mice treated with CL was prolonged (P <0.05), and thymus index and spleen index of them were raised (P <0.05). The inhibitory effect of CL on JEC cells was in concentration-and time-dependent manner. IC50 of CL on JEC cells was 0.05 microg mL(-1) in 96 hours. Growth curves showed right-shift with CL concentration increasing. The number of NBT positive JEC cells increased and the LDH concentration of cultured medium declined with CL increasing. Apoptosis of JEC cells with CL treated was induced in concentration-dependent manner, apoptotic percentage of CL 10 microg mL(-1) on JEC cells was 25.59% in 24 hours. CL arrested JEC cells in G2-M phase (P <0.05). CONCLUSION: CL has certainly anticancer effects in vivo and in vitro. Anticancer effect of CL in vivo was in relation to enhancing immune function of EAC mice; anticancer mechanisms of CL on JEC cells may be its direct cytotoxic effect, inducing cell apoptosis and inhibiting cell segmentation.

[Studies on the chemical constituents of Vaccinium iteophyllum].

7 compounds were isolated from the ethyl-acetate extract of Vaccinium iteophyllum Hance by using repeated silical gel column chromatography. These 7 compounds were identified by means of physico-chemical propertic and spectroscopic analysis as beta-sitosterol (I), ursolic acid (II), taraxerol (III), taraxerone (IV), friedelin (V), friedelinol (VI), 19,24-dihydroxyurs-12-en-3-one-28-oic acid (VII). The chemical constituents from this plant were reported here for the first time.

[Chemical constituents from Viscum nudum and their accelerating PC12 cell differentiation].

OBJECTIVE: To investigate chemical constituents of the stem of Viscum nudum and their bioacyivity. METHOD: The major chemical constituents were isolated from the AcOEt-solved part of ethanol-extract of the plant by column chromatography and the active screening test in vitro were taken out for looking for compounds to acccelerate PC12 cell differentiation. RESULT: 5 compounds were identified as eriodictyol (1), 5, 7-dihydroxy-3', 4'-dimethoxy flavanone (2), oleanolic (3), 5, 7-dihydroxychromone (4) and homeriodictyol (5) by spectral evidences, in which homeriodictyol (5) had acceleration differentiation to PC12 cell. CONCLUSION: All compounds were obtained from this plant for the first time, and bioactive constituent was observed in the AcOEt-solved part.

[Two new flavones from Fordia cauliflora of Yunnan].

AIM: To study the chemical constituents of the stem of Fordia cauliflora of Yunnan province. METHODS: The constituents were separated and purified by repeated silica column chromatography. The structures were elucidated by physical-chemical properties and spectroscopic data. RESULTS: Six compounds were isolated from the ethanol extract of the stem of Fordia cauliflora. They were identified as: 6-hydroxy-3-methoxy-6",6"-dimethylchromeno-(2", 3" : 7, 8)-flavone (1), 3-methoxy-6-(3-methyl-but-2-enyloxy)-6", 6"-dimethylchromeno-( 2", 3" : 7, 8)-flavone (2), 3, 6-dimethoxy-6", 6"-dimethylchromeno-( 2", 3" : 7, 8)-flavone (3), 7-hydroxy-4'-methoxyisoflavone (4), 7, 4'-dihydroxyisoflavone (5) and karanjin (6). CONCLUSION: Compounds 1 and 2 are new compounds. Compounds 3 -5 were isolated from the plant for the first time.

[Study on chemical constituents from Incarvillea arguta and their accelerating PC-12 cell differentiation].

OBJECTIVE: To study chemical constituents of Incarvillea arguta and their accelerating PC-12 cell differentiation. METHOD: The constituents were isolated and repeatedly purified on silica gel column chromatography, and were identified on the basis of physicochemical and spectroscopic analysis. The neurotrophic activity of different portion and all purified compounds from I. arguta was determined on the model of PC-12 cell. RESULT: Five compounds were isolated from BuOH portion of alcohol extraction of I. arguta. Their structures were identified as plantarenaloside (I), 5-hydroxy-4', 6 7-trimethoxy-flavone (II), 4', 5-dihydroxy-6, 7-dimethoxyflavone (III), 4', 5-dihydroxy-7-methoxyflavone (IV), 5-dydroxy-4', 7-dimethoxyflavone (V). CONCLUSION: Compound I is isolated from the plant for the first time and it has neurotrophic activity for PC-12 cell. Compounds II approximately V are isolated from the genus Incarvillea for the first time.

The synergistic activity of antibiotics combined with eight traditional Chinese medicines against two different strains of Staphylococcus aureus.

The ethanolic extracts of eight traditional Chinese medicines and four antibiotics were investigated for their combined effects on the resistance of Staphylococcus aureus (S. aureus) in vitro. Methicillin resistant S. aureus, which was isolated from patient and a standard strain, were used. Our results showed that there are differences in the effects of many combinations used on the standard strain and resistant strain of S. aureus. The ethanolic extracts of Isatis tinctoria, Scutellaria baicalensis and Rheum palmatum can improve the antimicrobial activity of four antibiotics we used.

[Studies on chemical constituents from the root of Mirablis jalapa].

OBJECTIVE: To investigate the chemical constituents from the root of Mirabilis jalapa. METHOD: Compounds were isolated from 75% ethanolic extract of the titled herb by silica gel column chromatography, and their structures were elucidated by physical and chemical evidences and spectroscopic analysis. RESULT: Four compounds were obtained and identified as (2, 5-dioxoimidazolidin-4-yl)-urea (1), glycerin monoeicosate (2), boeravinone (3) and beta-sitosterol (4). CONCLUSION: Compound (2) is a new compound, and compound (1) was obtained from this plant for the first time.

Antiplatelet aggregation activity of diterpene alkaloids from Spiraea japonica.

Six diterpene alkaloids with an atisine-type C(20)-skeleton isolated from the Chinese herbal medicines Spiraea japonica var. acuta and S. japonica var. ovalifolia, as well as eight derivatives of spiramine C and spiradine F were evaluated for the ability to inhibit aggregation of rabbit platelets induced by arachidonic acid, ADP, and platelet-activating factor (PAF) in vitro. The results showed that 12 of the 14 atisine-type diterpene alkaloids significantly inhibited PAF-induced platelet aggregation in a concentration-dependent manner, but had no effect on ADP- or arachidonic acid-induced aggregation, exhibiting a selective inhibition. It is the first report that C(20)-diterpene alkaloids inhibit PAF-induced platelet aggregation. However, spiramine C1 concentration-dependently inhibited platelet aggregation induced by PAF, ADP and arachidonic acid with IC(50) values of 30.5+/-2.7, 56.8+/-8.4 and 29.9+/-9.9 microM, respectively, suggesting a non-selective antiplatelet aggregation action. The inhibitory effect of spiramine C1 on arachidonic acid was as potent as that of aspirin. Primary studies of the structure-activity relationships for inhibition of PAF-induced aggregation showed that the oxygen substitution at the C-15 position and the presence of an oxazolidine ring in spiramine alkaloids were essential to their antiplatelet aggregation effects. These results suggest that the atisine-type alkaloids isolated from S. japonica are a class of novel antiplatelet aggregation agents.

Effects of spiramine T on antioxidant enzymatic activities and nitric oxide production in cerebral ischemia-reperfusion gerbils.

Spiramine T, an atisine-type diterpene alkaloid isolated from the Chinese herbal medicine Spiraea japonica var. acuta (Rosaceae), was shown to have neuroprotective effects on cerebral ischemia-reperfusion injury. In this study, the effects of spiramine T on antioxidant enzymes and nitric oxide production were evaluated in gerbils subjected to global forebrain ischemia (10 min) and reperfusion (5 days). Spiramine T (1.0 and 2.0 mg kg(-1) i.p.) markedly reduced the content of lipid peroxide (LPO), increased the glutathione peroxidase (GSH-PX) activity, and inhibited the increase of nitric oxidase (NOS) activity and nitric oxide production in the cortex during ischemia-reperfusion in gerbils. These results suggested that the neuroprotective effects of spiramine T were related to modulation of endogenous antioxidant enzymatic activities and reduction of the formation of nitric oxide.