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Abstract This study investigated the changes in the ingredients in Fallopia multiflora Thunb. Haraldson (FMT) root after processing it with different methods such as soaking, stewing, and steaming or combined methods. The total polyphenol, 2,3,5,4'-tetrahydroxystilben-2-O-ß-D-glucoside (THSG), and physcion contents in FMT products after processing were determined using high-performance liquid chromatography (HPLC) and ultraviolet-visible (UV-VIS) methods. The results demonstrated that the processing method and time significantly affected the contents of polyphenol, THSG, and physcion. The physcion and total polyphenol content increased or decreased during processing depending upon the processing time, while the THSG content gradually decreased with an increase in the processing time. The content of physcion (a substance that can cause liver toxicity) was analysed, and the suitable conditions for processing of the FMT products were determined as initial soaking in rice swill for 24 h and subsequent stewing with black beans and water for 12 h
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Fallopia multiflora/genetics , Methods , Chromatography, High Pressure Liquid/methods , Polyphenols/agonists , Liver/abnormalitiesABSTRACT
Aim To investigate the effect of PI3K/AKT/NF-κB signaling pathway in the treatment of physicion-8-O-β-D-monoglu-coside(PMG) on acute liver injury induced by carbon tetracloride(CCl 4) in mice . Methods Mice were randomly assigned into control group, model group, PMG low-dose, medium-dose and high-dose groups and Bifendate groups. After the continuous intervention with PMG for three days, CCl 4oil solution was intraperitoneally injected to establish acute liver injury mouse models, and samples were collected sixteen hours later. HE staining was used to observe the pathological changes of liver tissue. Hoechst 33342 staining was used to detect the number of apoptotic hepatocytes. Western blot was employed to detect the expression levels of caspase-3, PI3K, p-PI3K, AKT, p-Akt, IκB, p-IκB total protein and the nuclear protein NF-κB p65 in mouse liver tissue. The proportion of Th17 cells in mouse liver tissue was detected by FACS. Results After three days of PMG treatment, the pathological injury of liver tissue was relieved, the apoptosis of liver cells and the protein levels of caspase-3(P<0.01) were induced compared with model group.PMG could significantly decrease the nuclear expression of NF-κB p65 in the liver of mice with acute liver injury(P<0.05 or P<0.01). The phosphorylation levels of PI3K, AKT and IκB significantly decreased by PMG(P<0.05 or P <0.01). Otherwise, the proportion of Th17 cells in liver tissue was significantly reduced after PMG treatment(P<0.01). Conclusion PMG can alleviate CCl4 - induced acute liver injury through PI3K/AKT/NF-κB signaling pathway.
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Objective:To investigate the mechanism of physcion affecting the cell cycle and proliferation of prostate cancer DU145 cell line by regulating the expression of miR-380-3p.Methods:Prostate cancer DU145 cells were treated with 50 μg/mL physcion as physcion group, and normal cultured DU145 cells without any treatment were used as control group. Flow cytometry was used to detect DU145 cell cycle changes. MTT proliferation test was used to detect the proliferation of DU145 cells. quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-380-3p in DU145 cells. The bioinformatics software RNAhybrid was used to predict the target genes of miR-380-3p. qRT-PCR and Western blotting methods were used to detect the expression of miR-380-3p target gene. Measurement data were expressed as mean ± standard deviation ( ± s), t-test was used for comparison between two groups. Results:Compared with the control group, DU145 cells in the physcion group were blocked in the G 0/G 1 phase ( P<0.01), and the proliferation ability of DU145 cells was significantly inhibited ( P<0.05). The expression of miR-380-3p in DU145 cells in the control group and physcion group was 8.36 ± 1.42 and 1.08 ± 0.39, respectively. Physcion could promote the expression of miR-380-3p ( t=4.96, P<0.01). The functional target gene of miR-380-3p may be UHRF1. The relative expression levels of UHRF1 mRNA in DU145 cells in the physcion group and control group were 0.23±0.06 and 1.04±0.15, respectively. Compared with the control group, the expression of UHRF1 gene in DU145 cells in the physcion group was decreased ( t=4.55, P<0.01). Conclusion:Physcion can inhibit the proliferation of prostate cancer DU145 cells and induce G 0/G 1 block in DU145 cells, which may be closely related to the regulation of miR-380-3p.
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Objective: To establish HPLC characteristic chromatogram of standard pieces of Polygoni Multiflori Radix, and compare with the HPLC characteristic chromatograms of original pieces, raw materials and control materials of Polygoni Multiflori Radix. Method: HPLC analysis was carried out on a Waters BEH-C18 column (4.6 mm×250 mm, 5 μm) with the mobile phase of methanol(A)-0.1%phosphoric acid(B) for gradient elution (0-20 min, 15%-30%A; 20-35 min, 30%-40%A; 35-55 min, 40%-75%A; 55-75 min, 75%-100%A) at a flow rate of 0.8 mL·min-1. The detection wavelength was set at 270 nm and the column temperature was maintained at 30℃. The injection volume was 10 μL. Moreover, similarity and cluster analysis of HPLC characteristic chromatograms of four samples of Polygoni Multiflori Radix were performed. Result: HPLC characteristic chromatogram of standard pieces of Polygoni Multiflori Radix composed of seven peaks was established. The similarity was 0.999 between characteristic chromatograms of standard pieces and original pieces, which was better than similarities among characteristic chromatograms of raw materials, control materials and original pieces. There was no obvious difference on number of peaks between characteristic chromatograms of standard pieces and original pieces, while an obvious difference on number of peaks between chromatograms of control materials and original pieces was found. In addition, standard pieces, original pieces and control materials could generally gather into one class, while raw materials could generally gather into another class. Conclusion: Compared with the raw materials and control materials, the established HPLC characteristic chromatogram of standard pieces can better reflect the internal quality of original pieces, which can be used for the quality control of decoction pieces of Polygoni Multiflori Radix.
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Objective@#To study the effect of different proportions of piper chinaroot and rheum palmatum on the dissolution rate of five effective components (aloe emodin, emodin acid, emodin, emodin, emodin methyl ether).@*Methods@#The high performance liquid chromatography (HPLC) method was used to analyz the contents of five effective components of rheum palmatum in the extracts of different combination of piper chinaroot and rheum palmatum. The tests were carried out by Thermo C18 (250 mm×4.6 mm, 5 μm) by gradient elution with methanol and 0.1% phosphoric acid water solution as mobile phase at a flow rate of 1ml/min, the column temperature was 30 ℃ and the detection wavelength was 254 nm.@*Results@#The linear ranges of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 0.018 5-0.741 8, 0.017 9-0.717 8, 0.015 9-0.635 5, 0.054 2-2.167 2, 0.016 2-0.646 4 μg, respectively. The average recoveries of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 94.35%, 95.50%, 100.61%, 96.27%, 97.39%, and the RSDs were 1.81%, 1.99%, 2.84%, 2.71%, and 1.86%, respectively. Compared with rheum palmatumby single extract, the content of aloe emodin and emodin were increased by 5 proportions (3:1, 2:1, 1:1, 1:2, 1:3), while the content of emodin and emodin methyl ether were decreased.@*Conclusions@#The optimal compatibility proportion of piper chinaroot and rheum palmatum is 1:1.
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Objective: To establish a method for the determination of six components from Rumex chalepensis Mill.. Methods: The contents of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcion were simultaneously determined by HPLC. The mobile phase was methanol-0.1% formic acid, gradient elution, flow rate of 1 mL/min, column temperature of 25 ℃, injection volume of 5 μL, detected by Agilent Extend-C18 (250 mm × 4.6 mm, 5 μm) and diode array detector at 254 nm wavelength. Results: The content of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcionhad good linear relationship in the ranges of 208—3 120, 22.40—336.35, 178.9—2 908.8, 16.7—250.8, 104.4—1 566.0, 45.2—677.7 ng, respectively. The average recovery rates were 97.66%, 97.10%, 98.78%, 97.38%, 102.48%, and 95.51% (n = 6). The contents of chrysophanol-8-O-β-D-glucoside, emodin-8-β-D-glucoside, nepodin, emodin, chrysophanol, and physcion in 16 batches of R. chalepensis. were determined in the range of 0.6—7.1, 0—2.7, 1.0—6.5, 0.1—0.6, 0.7—4.3, and 0.1—0.4 mg/g, respectively. Sample contents of different growing years, harvesting dates, and plots were compared and analyzed. Two-year-old R. chalepensis. was collected in early spring or late summer and early autumn. The total content of six components was 12.2 mg/g, which was relatively high. Conclusion: The established method can be used for simultaneous determination of six components from R. chalepensis, and determine the harvesting time and season of R. chalepensis, which provides a scientific basis for the formulation of quality evaluation criteria of R. chalepensis.
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Objective: To study the chemical constituents from the rhizome of Tripterygium hypoglaucum. Methods: The concrete exacted by 95% ethanol from the rhizome of T. hypoglaucum was isolated and purified by chromatography on silica gel column chromatography, gel column chromatography, MPLC, etc. The structures of the chemical constituents were elucidated by means of NMR, MS, etc. Results: Twelve compounds were isolated and identified as 3β-hydroxy-4-hydroxymethyl-abieta-8,11,13-trien-7-one (1), 3β-acetoxy oleanolic acid (2), physcion (3), canophyllal (4), chrysophenol (5), quinone 21 (6), tripterifordin (7), triptobenzene B (8), 3,4-dimethoxyphenol (9), integracin A (10), celastrol (11), and β-sitosterol (12). Conclusion: Compound 1 is a new diterpene named tripterone, compounds 8-10 are isolated from this plant for the first time.
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To evaluate the hepatotoxicity risks of physcion on the basis of the bilirubin metabolism mediated by glucuronidation of UDP-glucuronosyltransferases 1A1(UGT1A1 enzyme). The monomers were added into the rat liver microsomes to test the hepatotoxicity by using bilirubin as UGT1A1 enzyme substrate, with apparent inhibition constant K_i as the evaluation index. Liver microsome incubation in vitro was adopted to initiate phase Ⅱ metabolic reaction and investigate the inhibitory effect of physcion. Then the phase Ⅰ and Ⅱ metabolic reactions were initiated to investigate the comprehensive inhibition of metabolites and prototype components. The results showed that when only the phase Ⅱ reaction was initiated, physcion directly acted on the UGT1A1 enzyme in a prototype form, exhibited weak inhibition and the inhibition type was mixed inhibition; When the phase Ⅰ and Ⅱ reactions were initiated simultaneously, the inhibitory effects of physcion on UGT1A1 enzyme became strong and the inhibition type was mixed inhibition, suggesting that physcion had phase Ⅰ and Ⅱ metabolic processes, and the metabolites had strong inhibitory effect on UGT1A1 enzyme. This experiment preliminarily proved that the metabolites of physcion may be the main components to induce hepatotoxicity.
Subject(s)
Animals , Rats , Chemical and Drug Induced Liver Injury , Emodin , Toxicity , Glucuronosyltransferase , Metabolism , Kinetics , Microsomes, LiverABSTRACT
OBJECTIVE:To develop a method for simultaneous determination of geniposide,baicalin,aloe-emodin,rhein, emodin,chrysophanol and physcion in Zhizi jinhua dispersible tablets. METHODS:HPLC method was adopted. The determination was performed on Dimonsil C18 column with mobile phase consisted of methanol-0.05%phosphoric acid(gradient elution)at the flow rate of 0.8 mL/min. The detection wavelength was set at 254 nm,and the column temperature was 25℃. The sample size was 20 μL. RESULTS:The linear ranges of geniposide,baicalin,aloe-emodin,rhein,emodin,chrysophanol and physcion were 0.0323-0.323 μg (r=0.9998),0.1374-1.374μg(r=0.9999),0.00372-0.0372μg(r=0.9997),0.0069-0.069μg(r=0.9995),0.00332-0.0332μg (r=0.9997),0.00864-0.0864 μg(r=0.9997) and 0.00122-0.0122 μg(r=0.9995),respectively. The limits of quantitation were 0.0321,0.1374,0.00372,0.0067,0.00330,0.00864,0.00122 μg,respectively. The limits of detection were 0.0095, 0.0041,0.0012,0.0020,0.0010,0.0026,0.0003 μg,respectively. RSDs of precision,stability and reproducibility tests were all lower than 3%. The average recoveries were 96.54%-99.52%(RSD=1.17%,n=6),97.23%-101.23%(RSD=1.36%,n=6), 97.22%-101.25%(RSD=1.83%,n=6),97.32%-100.23%(RSD=1.09%,n=6),97.99%-102.71%(RSD=1.73%,n=6), 96.99%-100.23%(RSD=1.21%,n=6),96.99%-103.01%(RSD=2.31%,n=6),respectively. CONCLUSIONS:The methods is simple and reproducible. It can be used for the content determination of 7 components in Zhizi jinhua dispersible tablets.
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Objective To develop an HPLC method to determine the contents of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B simultaneously in the aerial part of Rheum tanguticum Maxim.ex Balf. Methods The analysis was achieved with an Agilent ZORBAX SB-C18 analytic column (250 mm × 4.6 mm, 5 μm) by gradient elution of methanol-0.1％ phosphoric acid in water gradient elution system (0 min, 70％ B; 5 min, 65％ B;8–16 min, 60％ B; 18–23 min, 55％ B; 25–30 min, 45％ B; 32–39 min, 35％ B; 49–57 min, 24％ B; 65–72 min, 20％ B). The flow rate was 1.0 mL/min; detection wavelength was 254 nm; the column temperature was room temperature; the quantification used external standard method. Results The peak areas and concentrations of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B showed good linear relationship within a certain concentration range (r≥0.9995); the RSD of precision was 0.75％–1.17％; the RSD of repeatability was 0.99％–2.06％; the RSD of stability was 0.97％–1.76％; the average recoveries were 99.7％–100.4％. The results showed that there were differences in content between the root and aerial part of Rheum tanguticum Maxim.ex Balf. Conclusion HPLC method for simultaneous determination of contents of 6 contents of the aerial part of Rheum tanguticum Maxim.ex Balf can be used as the references for quality control.
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Objective:To establish the quality standard for Huayu pills. Methods:TLC was adopted to identify salvia miltiorrhi-za, rhubarb charcoal and panax notoginseng; the contents of tanshinoneⅡA , chrysophanol and physcion in Huayu pills were deter-mined by RP-HPLC with Wondasil C18 (250 mm × 4. 6 mm,5μm) as the analytical column, the mobile phase was composed of metha-nol-0. 1% phosphoric acid solution (85 ∶15), the flow rate was 1. 0 ml·min-1,the detection wavelength was 254 nm, the column temperature was 35℃, and injection volume was 10 μl. Results:The TLC spots were clear and well-separated without any negative in-terference; the calibration curve of tanshinoneⅡA , chrysophanol and physcion was linear over the range of 0. 212-2. 12 μg ( r =0. 999 9),0. 159-1. 59 μg(r=0. 999 9) and 0. 072 6-0. 726 μg(r=0. 999 9), the average recovery was 99. 55 %, 99. 56% and 100. 60%, and RSD was 1. 57%,2. 26% and 2. 28%(n=6), respectively. Conclusion: The method is fast and accurate with good specificity, and can be used for the quality control of Huayu pills.
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Objective: To establish a method for the simultaneous determination of geniposide, liquiritin, aloe-emodin, rhein, emodin, chrysophanol and physcion in Yinzhi Quhuang capsules by an HPLC wavelength switching method. Methods:A Waters Sun-fire C18(250 mm ×4.6 mm,5μm) column was used with the mobile phase of methanol-0.05% phosphoric acid solution (gradient elu-tion) at a flow rate of 1. 0 ml·min-1 , the column temperature was 25℃, the detection wavelength was 238 nm in 0-15 min for genipo-side and liquiritin, and 254 nm in 15-50 min for aloe-emodin,rhein, emodin, chrysophanol and physcion. The injection volume was 10μl. Results: The linear range was 0.13-3.18 μg·ml-1 (r =0.9998) for geniposide, 0.19-4.84 μg·ml-1(r =0.9997) for liquiritin, 0. 28-7. 02μg·ml-1(r=0. 9999) for aloe-emodin, 0. 13-3. 16μg·ml-1(r=0. 9999) for rhein, 0. 61-15. 27μg·ml-1 (r=0.9999) for emodin, 0.32-8.03 μg·ml-1(r=0.9999) for chrysophanol and 0.39-9.81 μg·ml-1(r=0.9999) for phy-scion. The average recovery was 98. 84%(RSD=0. 74%), 99. 34%(RSD=0. 86%), 99. 54%(RSD=0. 30%), 99. 56% (RSD=0. 80%), 99. 85% (RSD=0. 41%), 99. 57% (RSD=0. 70%) and 99. 64% (RSD=0. 30%) (n=9). Conclusion: The deter-mination method has the advantages of simple operation, high specificity, good reproducibility and accurate results, which can be used for the quality control of Yinzhi Quhuang capsules.
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OBJECTIVE:To study the effect and mechanism of physcion 8-O-β-glucopyranoside(PG)on the apoptosis of skin melanoma A375 cells. METHODS:After A375 cells were treated by PG with 0,10,20,50 μg/mL for 24,48,72 h,CCK-8 method was adopted to determine the survival rate of cells. After A375 cells were treated by PG with 0(control),20,50 μg/mL for 48 h,flow cytometry was used to detect the apoptosis rate of cells with membrane protein Ⅴ/propidium iodide (PI) double staining. Immunoblotting was used to detect the protein expressions of Caspase-3 and polyadenyl adenine diphosphate ribose poly-merase (PARP) and protein expressions of cytochrome C inside and outside mitochondria. After A375 cells were treated by PG with 0 (control),5,10 μmol/L for 48 h,enzyme substrate method was used to determine the activities of Caspase-8 and Cas-pase-9. RESULTS:PG can effectively decrease the survival rate of A375 cells. Compared with control,apoptosis rate of cells was obviously increased after treated by PG with 20,50 μg/mL(P<0.01);protein expressions of Caspase-3,PARP in cells and cyto-chrome C in cell matrix were obviously enhanced(P<0.05 or P<0.01);and protein expression of cytochrome C in mitochondria was obviously weakened(P<0.05 or P<0.01). Caspase-9 activity in cells was obviously enhanced after treated by PG with 5,10μmol/L(P<0.05 or P<0.01);and Caspase-8 activity had no obvious changes. CONCLUSIONS:PG can inhibit activity of A375 cells and promote its apoptosis,and its pro-apoptotic effects is achieved by destructing mitochondrial membrane potential and pro-moting cytochrome C outflow.
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OBJECTIVE:To study the effect and mechanism of physcion 8-O-β-glucopyranoside(PG)on the apoptosis of skin melanoma A375 cells. METHODS:After A375 cells were treated by PG with 0,10,20,50 μg/mL for 24,48,72 h,CCK-8 method was adopted to determine the survival rate of cells. After A375 cells were treated by PG with 0(control),20,50 μg/mL for 48 h,flow cytometry was used to detect the apoptosis rate of cells with membrane protein Ⅴ/propidium iodide (PI) double staining. Immunoblotting was used to detect the protein expressions of Caspase-3 and polyadenyl adenine diphosphate ribose poly-merase (PARP) and protein expressions of cytochrome C inside and outside mitochondria. After A375 cells were treated by PG with 0 (control),5,10 μmol/L for 48 h,enzyme substrate method was used to determine the activities of Caspase-8 and Cas-pase-9. RESULTS:PG can effectively decrease the survival rate of A375 cells. Compared with control,apoptosis rate of cells was obviously increased after treated by PG with 20,50 μg/mL(P<0.01);protein expressions of Caspase-3,PARP in cells and cyto-chrome C in cell matrix were obviously enhanced(P<0.05 or P<0.01);and protein expression of cytochrome C in mitochondria was obviously weakened(P<0.05 or P<0.01). Caspase-9 activity in cells was obviously enhanced after treated by PG with 5,10μmol/L(P<0.05 or P<0.01);and Caspase-8 activity had no obvious changes. CONCLUSIONS:PG can inhibit activity of A375 cells and promote its apoptosis,and its pro-apoptotic effects is achieved by destructing mitochondrial membrane potential and pro-moting cytochrome C outflow.
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AIM To establish an HPLC method for determining the contents of three constituents in Polygoni multiflori Radix Praeparata in plasma of atherosclerosis rats.METHODS After the rats were intragastrically administrated with Polygoni multiflori Radix Praeparata CMC-Na solution,the plasma was collected.The HPLC analysis was carried out on a 30 ℃ thermostatic Hypersil C1s column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.03％ phosphoric acid flowing at 0.9 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.DAS2.0 software was applied to drawing concentration-time curves and calculating pharmacokinetic parameters.RESULTS Stilbene glucoside,emodin and physcion showed good linear relationships within the ranges of 61.25-6 125 μg/L (r =0.999 8),12.6-3 150 μg/L (r =0.999 3) and 24.1-6 030 μg/L (r =0.999 5),respectively.The method recoveries were 99.5％-105.8％ with the RSDs of 1.3％-3.3％,while the extraction recoveries were 87.2％-96.3％ with the RSDs of 3.2％-5.9％.The pharmacokinetic behaviors of three constituents all accorded with two-compartment model,but their contents in plasma were much lower than those in medicinal material.CONCLUSION The bioavailabilities of stilbene glucoside,emodin and physcion are relatively low in plasma of atherosclerosis rats,which may be related to constituents' intestinal absorption after intragastric administration with Polygoni multiflori Radix Praeparata.
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Objective: To analyze the main components in different parts of Rheum palmatum. Methods: The anthraquinones, soluble polysaccharides, cellulose, and mineral elements in the taproots, root heads, fibrous roots, root barks, petioles and leaves were detected by HPLC, UV, Weende, and ICP-AES. Results: The contents of aloe-emodin, rhein, emodin, chrysophanol, and physcion in taproots, root heads, fibrous roots, and root barks were 3.22-4.33, 1.33-2.32, 3.21-3.68, 3.22-3.76 mg/g, 0.77-1.36, 2.46-2.52, 1.16-1.46, 1.02-1.21 mg/g, 0.27-0.39, 0.28-0.34, 0.30-0.42, 0.31-0.67 mg/g, 2.85-3.70, 2.78-3.01, 4.02-4.81, 4.05-4.72 mg/g and 1.88-2.44, 1.82-2.01, 2.48-3.02, 3.61-4.46 mg/g, respectively. The contents of aloe-emodin, rhein, and emodin in leaves were 0.56-1.07, 0.45-0.69, 1.41-1.91 mg/g. The soluble polysaccharides in the taproots, petioles and leaves were 9.76%-10.42%, 5.76%-7.63%, and 3.50%-5.72%. The cellulose contents in petioles and leaves were 15.54% and 10.20%. Ca was the most abundant with 88.53 mg/g in leaves, followed by K with 32.42 mg/g, Mg with 12.93 mg/g, Al with 1.22 mg/g, and Fe with 1.17 mg/g. In the petioles, Ca with 80.60 mg/g and K with 28.73 mg/g were higher than those in roots with 21.08 and 14.09 mg/g. Na with 2.66 mg/g was also higher than that in roots with 0.26 mg/g and in leaves with 0.57 mg/g. Conclusion: The types and contents of anthraquinones in roots are higher than those in petioles and leaves, with the understanding of traditional medicinal parts. Emodin in leaves is five times as those in roots, petioles, and leaves, and also contains a certain amount of cellulose and soluble polysaccharide component, a wide variety of elements. From above analysis, the petioles and leaves could be deeper utilized.
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Objective: To study the chemcial constituents from Melicope ptelefolia. Methods: The constituents were isolated and purified by chromatography with silica gel, Sephadex LH-20, and recrystallization. The structures were elucidated by sepectroscopic anaylsis (NMR and ESI-MS). Results: Twelve compounds were isolated from petroleum ether and ethyl acetate soluble fraction and identified as: physcion (1), stigmata-3,5-dien-7-one (2), β-sitostenone (3), 24(R)-24-ethyl-5α-cholestane-3β,5α,6β-triol (4), dihydroxanthyletin (5), methyl icosanoate (6), emodin (7), (+)-marmesin (8), rudicoumarin C (9), 3-(2',3'-dihydroxy) isopentyl-7- hydroxycoumarin (10), and 4-methoxy-2(1H)-quinolinone (11). Conclusion: Compounds 1-11 are isolated from M. ptelefolia for the first time. Compound 10 is a new compound named pteleifolosin D.
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Objective To analyze the main chemical constituents and their contents in aqueous extract of Polygoni Multiflori Radix (PMR, root of Polygonum multiflorum), and to elucidate the effects of aqueous extract of PMR and its main constituents on the expression of the mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells. Methods The main chemical constituents and their content in aqueous extract of PMR were determined by HPLC. The cytotoxicity of aqueous extract of PMR and its main constituents on L02 cells was determined by MTT assay. The mRNA expression of CYP1A2, CYP2C9, and CYP2E1 in L02 cells were detected by quantitative real-time PCR. Results There were four main well-separated chromatographic peaks standing for tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside, emodin and physcion in aqueous extract of PMR. The content of thesecomponents in aqueous extract of PMR was (1.14 ± 0.03)%, (0.106 9 ± 0.001 6)%, (0.010 8 ± 0.000 9)%, (0.003 55 ± 0.000 19)%, respectively. The cytotoxicity of aqueous extract of PMR and emodin on L02 cells at 24 h was dose-dependent, and the concentration of 50% inhibition was 7.290 mg/mL and 0.082 mmol/L respectively. Tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside and physcion did not show significant cytotoxicity on L02 cells in the experimental concentrations. Aqueous extract of PMR and emodin significantly inhibited the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells. Emodin-8-O-β-D-glucoside inhibited the expression of mRNA of CYP1A2 and CYP2C9. Tetrahydroxy stilbene glucoside inhibited the expression of mRNA of CYP1A2 but activated the expression of mRNA of CYP2C9. Physcion inhibited the expression of mRNA of CYP1A2 and CYP2C9 in a dose-dependent manner, but inhibited the expression of mRNA of CYP2E1 in low concentration and activitated the expression of mRNA of CYP2E1 in high concentration. Conclusion The inhibition of aqueous extract of PMR on the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells is the combined effect of all components in it. The main four components all inhibit the expression of mRNA of CYP1A2. The anthraquinone is the main component inhibiting the expression of mRNA of CYP2C9. The free anthraquinone is the main component inhibiting the expression of mRNA of CYP2E1.
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Objective To develop a method of quantitative analysis of multi-components by single marker(QAMS) for determination of effective components of naphthopyrone glycosides and anthraquinone aglycone in Cassia tora L.Methods The separation was performed on a Sunfire C18 column (4.6 mm ×250 mm,5 μm), and column temperature 30 ℃.Aetonitrile-0.1% H3PO4 was used as the mobile phase with gradient elution.The detection wavelength was at 278 nm and 284 nm.Relative correction factor by determination of Between aurantio-obtusin and rubrofusarin-gentiobioside or chrysophanol or physcion.the method was evaluated for reproducibility, and the difference between calculated and measured values was compared. Results Rubrofusarin-gentiobioside, aurantio-obtusin, chrysophanol and physcion respectively 0.0503-1.5078, 0.3197-9.5899, 0.5070-15.2108, 0.4027-12.0814μg showed a good linear relationship, and the regression equation is Y =4.95X +2.01(R2 =0.9998),Y =1.03X +0.03(R2 =0.9999),Y=3.98X-0.12(R2 =0.9993),Y =4.81X +0.26(R2 =0.9996).The quantitative results of six batches of Cassia tora L by QAMS was basically consistent with that by external standard method.Conclusion The QAMS method was reliable and accurate, which might be used for the quality control of Cassia tora L.
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OBJECTIVE: To study the chemical constituents of the fruits of Embelia laeta. METHODS: The compounds were isolated and purified by MCI, medium pressure TLC on silica gel column, ODS column chromatography, semi-preparative HPLC, and so on. The structures were elucidated on the basis of various modern spectroscopic techniques using physical, chemical properties and spectral data. RESULTS: Fifteen compounds were isolated and identified as nantenine(1), oxonantenine(2), physcion(3), syringic acid(4), vanillic acid(5), stigmast-4-ene-3,6-dione(6), (+)-lyoniresinol(7), 75,85-threo-4,7,9,9'-tetrahydroxy-3,3'-dimethoxy-8-O-4'-neolignan(8), 1,3-dihydroxylpropyl-(9Z,12Z)-octadeca-9,12-dienate(9),(22E)-5a,8a-epidioxyergosta-6,22-dien-3b-ol(10), dihydroxyisoechinulin A(ll), hydroxybenzoic acid(12), stigmasterol(13), sitosterol(14), and daucosterol(15). CONCLUSION: Compounds 1,2, and 5-13 are isolated from the genus Embelia for the first time, and compounds 1-15 are isolated from this plant for the first time.