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Objective To screen the active components of total flavonoid extracts of Sarcandra glabra to promote megakaryocyte differentiation.Methods(1)A model of megakaryocyte differentiation disorder was established by co-culturing human megakaryocytic leukaemia cells(Dami)with human bone marrow stromal cells(HS-5)as an evaluation system,and the experimental groupings were as follows:the Dami group(Dami),the control group(Dami+HS-5),and the PMA group[Dami+HS-5+5 ng·mL-1 foprolol 12-tetradecanoate 13-acetate(PMA)],and model group[Dami+HS-5+1%rabbit anti-rat platelet serum(APS)+5 ng·mL-1 PMA]were cultured for 48 hours.The expressions of megakaryocyte differentiation and maturation surface marker molecules,CD41a and CD61 were detected by flow cytometry.(2)Forty-nine SD male rats were randomly divided into blank plasma group,15-minute group,30-minute group,60-minute group,90-minute group,120-minute group,and 240-minute group,with 7 rats in each group.The rats in each administration group were gavaged with 1.26 g·kg-1 of total flavonoids extracts of Sarcandra glabra,and blood was collected at six set time points(15,30,60,90,120,240 minutes)for the preparation of time-dependent serum-containing plasma of total flavonoids extracts of Sarcandra glabra.(3)Ultra-high performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry(UHPLC-Q-TOF/MS)was used to analyze the plasma of the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra,and the peak area was used to construct a matrix(X-matrix)of the amount of chemical composition change over time in the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra.The collected time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra at six different time points was used to intervene in the model of megakaryocyte differentiation and maturation disorder,and the expression of cell surface molecules CD41a and CD61 was detected by flow cytometry to construct the matrix of effect of time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra(Y-matrix).(4)After the data of X and Y matrices were standardized,partial least squares(PLS)was used to calculate and analyze the quantitative and qualitative effect relationship,and variable importance for projection(VIP)>1 was used as the threshold to screen the effect components related to the changes of cell surface molecules CD41a and CD61,and chemical composition identification,as the potential effector components in the total flavonoid extracts of Sarcandra glabra were used to promote the differentiation of megakaryocytes,and finally the regression evaluation system was used to verify the efficacy of its medicinal effect.Results(1)Compared with the Dami group,the expression level of CD41a on the surface of Dami cells in the control group was significantly increased(P＜0.05).Compared with the control group,the expression levels of CD41a and CD61 on the surface of Dami cells in the PMA group were significantly increased(P＜0.01).Compared with the PMA group,the expression levels of CD41a and CD61 on the surface of Dami cells in the model group were significantly reduced(P＜0.01).(2)Compared with the blank plasma group,the expression levels of the molecules CD41a and CD61 on the surface of Dami cells at each time point of 15,30,60,90,120,and 240 minutes were significantly increased(P＜0.01),and the expression levels of CD41a and CD61 were both highest in the 30-minute group.The potential effective components with VIP value greater than 1 were screened out in the positive and negative ion mode,and 540.3638@12.25 and 559.2991@11.53 were selected for pharmacodynamic verification.559.2991@11.53 was identified as daucosterol(Dau),540.3638@12.25 was identified as rosmarinic acid 4-O-β-D-glucoside(Ros).After Ros and Dau intervened in the megakaryocyte differentiation and maturation disorder model respectively,the expression levels of CD41a and CD61 on the surface of Dami cells in the low-,medium-and high-dose groups(40,60 and 80 μg·mL-1)of Ros and Dau were significantly increased compared with the model group(P＜0.05,P＜0.01).Conclusion Ros and Dau may be the active components of the total flavonoids extracts of Sarcandra glabra to promote the differentiation of megakaryocytes.
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Objective: To study the chemical constituents of Lonicerae Japonicae Caulis produced in Shandong Province. Methods: Compounds were isolated and purified by repeated column chromatographies including silica gel, MCI-gel resin, Sephadex LH-20, and so on. Their structures were elucidated by spectral data, including extensive 1D and 2D NMR techniques. Results: Four compounds were obtained and identified as 11,14,15-trihydroxy-12-octadecenoic acid methyl ester (1), ochnaflavone (2), β-daucosterol (3), and loganin (4). Conclusion: Compound 1 is a new organic acid named as loniceric acid ester, and compound 2 is isolated from Lonicerae Japonicae Caulis for the first time.
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Objective: To establish the UPLC fingerprint of triterpenoids in Guizhi Fuling Capsule (GFC) and give a new method for quality control. Methods: UPLC was used on an Agilent Zorbox Eclipse Plus HD C18 (100 mm × 2.1 mm, 1.8 μm) column with the gradient elution solvent system composed of acetonitrile-0.1% H3PO4 water solution as mobile phase, the flow rate was 0.2 mL/min, the column temperature was 30℃, and the detection wavelength was 210 nm. The common peaks were identified by Q-TOF/MS. Results: The UPLC fingerprints of triterpenoids in GFC were established. Totally 20 common peaks were selected as the fingerprint peaks of GFC, of which a total of 13 mutual peaks (1-7, 10, 12, and 14-17) from Poria, the peaks of number 8 and 11 were from Cinnamomi Ramulus, the peak of number 9 was from Paeoniae Radix Alba and Moutan Cortex, and the peak of number 13 was from Persicae Semen, Paeoniae Radix Alba, Moutan Cortex, and Cinnamomi Ramulus. The similarity among the fingerprint of 10 batches of GFC samples were 0.90. Fifteen chemical components were identified by UPLC-Q-TOF/MS, which were 1-hederagenin, 2-dehydrotumulosic acid, 3-tumulosil acid, 4-polyporenic acid C, 6-3-epidehydrotumulosic acid, 7-poricoic acid D, 9-α-linolenic acid, 10-dehydropachymic acid, 11-oleanolic acid, 12-pachymic acid, 13-linolic acid, 15-methylcis-9-hexadecenoate, 16-palmitic acid, 17-palmitic acid ethyl ester, and 18-daucosterol. Conclusion: The established UPLC fingerprint has desirable precision, reproducibility, and stability, and could be applied to the quality control of GFC.
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Objective: To study the chemical constituents from the inflorescence bracts of Arctii Fructus. Methods: The compounds were isolated and purified by the methods of silica gel column chromatography, HPLC, and recrystallization, and the structures were elucidated by the means of spectral analysis. Results: Twelve compounds were isolated and identified as daucosterol (1), isofouquierol (2), (22E)-5α, 8-epidioxyergosta-6, 22-dien-3β-ol (3), 3β-hydroxy-21, 22-epoxyursa-20(30)-en (4), 3β, 21β-dihydroxy-20(30)-en-taraxastane (5), oleanolic acid (6), arctigenin (7), carthamogenin (8), caffeic acid (9), 4(14)-eudesmene-8α, 11-diol (10), monogynol A (11), and lupeol (12). Conclusion; Compounds 2-3, 5, 6, 10-11 are obtained from the plants of Arctium L. for the first time, and compound 12 is isolated from the inflorescence bracts of Arctii Fructus for the first time.
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Objective: To research the chemical constituents of Rabdosia pseudoirrorata. Methods: The chemical constituents were isolated and purified by silica gel column chromatography and their structures were determined by physicochemical properties and spectral analyses. Results: Seven compounds were isolated from the ethyl acetate extract fraction in 95% ethanol extract of R. pseudoirrorata and identified as: pseurata I (1), pseurata H (2), pseurata F (3), pseurata B (4), β-sitosterol (5), ursolic acid (6), and β-daucosterol (7). Conclusion: Compound 1 is a new compound, compounds 5-7 are first obtained from R. pseudoirrorata.
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Objective: To study the chemical constituents with anti-oxidative activity from the fruits of Illicium micranthum. Methods: DPPH Radical scavenging assay was used for screening the active fractions. The chemical constituents were isolated and purified by various column chromatographic methods such as silica gel and Sephadex LH-20. The structures were identified by physicochemical properties and spectral analyses. Results: The ethyl acetate layer of methanol extract from the fruits of I. micranthum showed anti-oxidative activity, from which 14 compounds were separated. Their structures were identified as gynurenol (1), β-elemol (2), epi-carrisone (3), 6-hydroxy-eudesm-4(14)-ene (4), β-eudesmol (5), trans-eudesmane-4, 11-diol (6), 1R, 2R, 4R-trihydroxy-p-menthane (7), 2-isopropenyl-5-methyl-cyclohexanol (8), (-)-isopulegol (9), 4-isopropenyl-1-methyl-cyclohexanol (10), palmitic acid (11), triacontanoic acid (12), p-hydroxy-benzoic acid (13), and β-daucosterol (14). Conclusion: Except for compound 14, all the other compounds are isolated from the fruits of I. micranthum for the first time, while compounds 1-4, 7, and 8 are isolated from the plants in Illiciaceae family for the first time.
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Objective: To study the chemical constituents from the whole plant of Actinostemma lobatum. Methods: The compounds were isolated and purified by various column chromatographies. Their structures were elucidated by means of chemical evidences and spectral analyses (MS, 1H-NMR, and 13C-NMR). Results: Sixteen compounds were isolated and identified as actinostemmoside A (1), actinostemmoside C (2), actinostemmoside D (3), actinostemmoside E (4), cucurbitacin E (5), β-sitosterol (6), daucosterol (7), α-spinasterol (8), α-spinasterol-3-O-β-D-glucopyranoside (9), 22, 23-dehydroxy-α-chondrillasterone (10), quercetin (11), kaempferol (12), rutin (13), quercitrin (14), kaempferol-3-O-α-L-rhamnopyranosyl (1→6)-β-D-glucoypyranoside (15), and isoquercitrin (16). Conclusion: Compounds 5-10 are obtained from this genus for the first time.
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The extraction of the bark of Parkinsonia aculeata, family Leguminosae was carried out using benzene, chloroform, ethanol and distilled water in succession. The presence of alkaloids, flavonoids, tannins, steroids, and reducing sugars was confirmed during preliminary phytochemical screening. The extracts were evaluated for antidiabetic activity. The chloroform and ethanol extracts showed significant antidiabetic activity.
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A espécie Bromelia antiacantha Bertol. (Bromeliaceae), conhecida como caraguatá ou gravatá, foi avaliada quanto à sua propriedade hemolítica frente a hemácias de sangue de carneiro. O extrato aquoso dos frutos apresentou atividade hemolítica a partir da diluição de 0,85%, enquanto que o extrato aquoso das folhas apresentou hemólise a partir da diluição de 0,90%. Tal atividade pode estar relacionada à presença de saponinas, uma vez que o estudo químico do extrato metanólico das folhas resultou no isolamento da saponina daucosterol, um fitoesterol glicosilado
The specie Bromelia antiacantha Bertol. (Bromeliaceae), popularly known as caraguatá or gravatá, was evaluated as for its hemolytic property against sheep red blood cells. The aqueous extract of the fruits presented hemolytic activity starting from the dilution of 0,85%, while the aqueous extract of the leaves presented hemolysis starting from the dilution of 0,90%. The activity should be related to the presence of saponins, once the chemical study of the leaves methanolic extract resulted in the isolation of the saponin daucosterol, a glycosylated phytosterol.
Subject(s)
Bromeliaceae , EryngiumABSTRACT
Objective:To study the chemical constituents of the peels collected from Citrus changshan-huyou Y.B.Chang,and further screen the bioactive components as the lead structures.Methods: These compounds were isolated by repeated flash column chromatography on silica gel and Sephadex LH-20.The structures of isolated compounds were elucidated by using IR,EIMS,and NMR analyses.Results: One novel compound along with nine known compounds were obtained and identified as huyoujiasu(Ⅰ),3-hydroxy-4-methoxybenzoic acid(Ⅱ),3,4-dihydroxybenzoic acid(Ⅲ),fatty acid(Ⅳ),gly-ceride(Ⅴ),6,7-dimethoxycoumarin(Ⅵ),6′,7′-dihydroxybergamottin(Ⅶ),daucossterol A(Ⅷ),daucossterol B(Ⅸ) and huyou-triterpenoid(Ⅹ).Conclusion: One new compound,Huyoujiasu,was obtained from these peels by repeated column chromatography,and the other known compounds were isolated from this fruit for the first time as well.
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Objective To study the chemical constituents in Sparganium stoloniferum. Methods The compounds were isolated by repeated silica gel and Sephadex LH-20 chromatographies. The structures of isolated compounds were identified by analysis of their spectral data and chemical reactions. Results Nine compounds were isolated and their structures were identified as daucosterol palmitate (Ⅰ), ?-sitosterol palmitate (Ⅱ), 24-methylenecycloartanol (Ⅲ), 6, 7, 10-trihydroxy-8-octadecenoic acid (Ⅳ), vanillic acid (Ⅴ), p-hydroxybenzaldehyde (Ⅵ), 2,3-dihydroxypropyl palmitate (Ⅶ), 5-hydroxymethyl-2-furaldehyde (Ⅷ), ?-sitosterol (Ⅸ). Conclusion Compounds Ⅰ-Ⅲ and Ⅴ-Ⅷ are obtained from the plants of Sparganium L. for the first time.
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Objective To investigate the chemical constituents in the roots of Boehmeria nivea.Methods The constituents were isolated by repeated column chromatography and their structures were elucidated by chemical properties and spectroscopic analyses.Results Seven compounds were isolated and their structures were identified to be daucosterol-10,13-eicosdienoate(Ⅰ),daucosterol(Ⅱ),?-sitosterol(Ⅲ),olein(Ⅳ),betulinic acid(Ⅴ),oleanolic acid(Ⅵ),19?-hydroxyursolic acid(Ⅶ).Conclusion Compound Ⅰ is a new compound named niveain A,compound Ⅳ is obtained from the plants of Boehmeria Jacq.for the first time.