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ObjectiveThrough the correlation analysis between intestinal absorption profile and inhibition of macrophage foaming， the pharmacodynamic components of Zhuriheng dripping pills（ZRH） were explored to provide a basis for establishing its quality standard. MethodIntestinal absorption fluids with 0， 5， 10， 15， 20 times clinical equivalent doses were prepared by a rat everted gut sac（EGS）， and the oxidized low density lipoprotein（ox-LDL）-induced RAW264.7 macrophage foaming model was used to investigate the effect of intestinal absorption fluid with different doses on the accumulation of lipids in RAW264.7 cells by oil red O staining and cholesterol content determination， and to screen for the optimal dose. Ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used to analyze and identify intestinal absorption fractions of ZRH intestinal absorption fluids， and partial least squares-discriminant analysis（PLS-DA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were performed on different doses of ZRH intestinal absorption fluids using SIMCA 13.0 with peak area as the independent variable and the pharmacodynamic indicators as the dependent variables to screen the compounds with variable importance in the projection（VIP） value>1.0 as contributing components， and Pearson correlation analysis was used to determine the spectral effect relationship， determined the compounds and positive correlation with pharmacodynamic were as active ingredients. Molecular docking was used to verify the binding energy of peroxisome proliferator-activated receptor α（PPARα）， PPARγ， PPARβ， human retinoid X receptor α（RXRA） and nuclear transcription factor-κB（NF-κB） with the active ingredients in ZRH intestinal absorption fluids. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was performed to detect the mRNA levels of PPARγ， scavenger receptor A1（SRA1） and adenosine triphosphate-binding cassette transporter A1（ABCA1） in RAW264.7 cells， Westen blot was used to detect the expression level of PPARγ protein in RAW264.7 cells， and enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of interleukin（IL）-1β and NF-κB in RAW264.7 cells. ResultAccording to the results of oil red O staining and cholesterol content determination， the ZRH intestinal absorption fluids could significantly reduce macrophage foaming， and intestinal absorption fluids with 15， 20 times clinical equivalent doses had the best effect， the 15-fold ZRH intestinal absorption fluid was finally determined as the study subject. Spectral effect relationship showed that 52 corresponding peaks in the ZRH-containing intestinal fluid were positively correlated with the efficacy， including organic acids， phenylpropanoids， iridoids， flavonoids， bile acids， coumarins and chromones. Target validation results showed that 86.9%-96.2% of the total components processed good binding activities with the key targets of PPARα， PPARγ， PPARβ， RXRA and NF-κB， and the docking energy values were all less than -6.0 kcal·mol-1（1 cal≈4.19 J）. The results of validation showed that， compared with the normal group， the model group showed a significant increase in the levels of SRA1 and PPARγ mRNA expression， a significant decrease in ABCA1 mRNA expression， a significant increase in the level of PPARγ protein expression， and a significant increase in the levels of IL-1β and NF-κB（P<0.01）， compared with the model group， the 15-fold intestinal absorption fluid group showed a significant decrease in the levels of SRA1 and PPARγ mRNA expression（P<0.05， P<0.01）， ABCA1 mRNA expression level was significantly up-regulated， the levels of IL-1β and NF-κB were significantly reduced（P<0.01）， and PPARγ protein expression level was significantly reduced（P<0.05）. ConclusionThis study identifies 52 components and their metabolites in ZRH intestinal absorption fluid that are positively correlated with the inhibition of macrophage foaming， which may be related to the regulation of the PPARs pathway in cells and the reduction of the levels of inflammatory factors， and can provide a reference for the quality control and clinical application of ZRH.
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OBJECTIVE To study the intervention effect and metabolic mechanism of Mongolian medicine Echinops sphaerocephalus extract on D-galactose-induced osteoporosis. METHODS Thirty-six 12-week-old male Wistar rats were selected and randomly divided into blank group， model group， Gushukang group， E. sphaerocephalus high-dose， medium-dose and low- dose groups， with 6 rats in each group. Except for blank group， other groups were intraperitoneally injected with D-galactose at 120 mg/kg per day. After 8 weeks of continuous injection， E. sphaerocephalus high-dose， medium-dose and low-dose groups were given drugs intragastrically at dose of 878， 439， 219.5 mg/kg， respectively. Gushukang group was given Gushukang 105.1 mg/kg intragastrically， once a day， for consecutive 8 weeks. After last administration， blood was collected from the abdominal aorta. Enzyme-linked immunosorbent assay was used to measure the contents of bone metabolism indexes [hydroxyproline （HYP）， alkaline phosphatase （ALP）] and oxidative stress indexes [total antioxidant capacity （TAOC）， superoxide dismutase （SOD）， malondialdehyde （MDA）] in serum of rats. Positron emission tomography/computedtomography （PET/CT） was used to analyze the changes of bone microstructure in right tibia bone. Meanwhile， metabolomic technology was used to study the regulation effect of E. sphaerocephalus on osteoporosis model rats. RESULTS Compared with blank group， HYP， ALP， MDA， ratio of bone surface to bone volume （BS/BV）， and trabecular separation （Tb·Sp） in model group were significantly increased （P＜0.05）， while TAOC， SOD， bone mineral density （BMD）， bone volume fraction （BVF）， trabecular E-mail：Xpfdc153@163.com thickness （Tb·Th） and trabecular number （Tb·N） were significantly decreased （P＜0.05）. Compared with model group， above indexes of administration groups were all reversed to different extents. The results of metabonomics study showed that after intervened with the extract of E. sphaerocephalus， 18 metabolites such as arachidonic acid， phenylalanine， tyrosine， tryptophan， isoleucine and uric acid in the serum of rats changed significantly， involving 15 metabolic pathways such as arachidonic acid， phenylalanine and tyrosine， of which arachidonic acid metabolism， phenylalanine metabolism and tyrosine metabolism were the main influencing pathways. CONCLUSIONS E. sphaerocephalus extract can effectively improve D-galactose-induced oxidative stress and the deterioration of bone microstructure， which interferes with metabolic pathways such as arachidonic acid metabolism and amino acid metabolism.
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OBJECTIVE To optimize the extraction technology of modified Tabusen- 2（MT-2），and to investigate inhibitory effects of the extract obtained by the optimal technology on osteoclast differentiation. METHODS The index components of MT- 2 process optimization were selected by using network pharmacology. Based on single factor tests ，the extraction technology of MT- 2 was optimized by Box-Behnken design-response surface methodology according to the comprehensive score of contents of above index components ，and then validated. RAW 264.7 cells were induced by receptor activator of nuclear factor-κB ligand（100 ng/mL） to prepare osteoclast differentiation model. Inhibitory effects of MT- 2 extract（18.6，37.2，74.4 ng/mL）obtained by the optimal technology on osteoclast differentiation were investigated. RESULTS The index components screened by network pharmacology included chlorogenic acid ，terpineol diglucoside ，isochlorogenic acid A ，1，5-dicaffeoylquinic acid ，hydroxysafflower yellow A ， ginsenoside Rg 1 and ginsenoside Rb 1. The optimal extraction technology of MT- 2 was ethanol volume fraction of 60％ ，the solid-liquid ratio of 1 ∶ 14（g/mL），extraction time of 94 min and extraction times of twice. The average comprehensive score obtained by the three validation experiments was 95.50，and the relative error with the predicted value （95.75）was －0.26％. Compared with osteoclastic differentiation model cells ，the cells treated with MT- 2 extract prepared by the optimal technology were mostly mononuclear round cells ，and the number of osteoclasts decreased significantly （P＜0.05），its inhibitory effects tended to strengthen with the increase of drug concentration. CONCLUSIONS The optimal extraction technology of MT- 2 is stable and feasible. Obtained extract can inhibit osteoclast differentiation.
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OBJECTIVE:To establish a method for simultaneous determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.METHODS:HPLC method was adopted.The determination was performed on Thermo scientific ODS-2 Hypersil column with mobile phase consisted of acetonitrile-0.5％ phosphoric acid (gradient elution) with flow rate of 1.0 mL/min.The detection wavelength was set at 237 nm,and column temperature was 25 ℃.The sample size was 20 μL.RESULTS:The linear ranges of liquiritin and glycyrrhizic acid were 48-480 μg(r=0.999 9) and 90-900 μg(r=0.999 9),respectively.The limits of quantitation were 0.55,2.10 μg/mL,and the limits of detection were 0.16,0.62 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 95.08％-97.58％ (RSD=0.93％,n=6)and 95.86％-99.89％ (RSD=1.67％,n=6),respectively.CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the content determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.
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OBJECTIVE:To establish a method for simultaneous determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.METHODS:HPLC method was adopted.The determination was performed on Thermo scientific ODS-2 Hypersil column with mobile phase consisted of acetonitrile-0.5％ phosphoric acid (gradient elution) with flow rate of 1.0 mL/min.The detection wavelength was set at 237 nm,and column temperature was 25 ℃.The sample size was 20 μL.RESULTS:The linear ranges of liquiritin and glycyrrhizic acid were 48-480 μg(r=0.999 9) and 90-900 μg(r=0.999 9),respectively.The limits of quantitation were 0.55,2.10 μg/mL,and the limits of detection were 0.16,0.62 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 95.08％-97.58％ (RSD=0.93％,n=6)and 95.86％-99.89％ (RSD=1.67％,n=6),respectively.CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the content determination of liquiritin and glycyrrhizic acid in Wuwei shaji granules.
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Echinops latifolius Tausch is a plant of compositae echinops, whose dried roots are used as traditional Chinese medicine Yuzhou Loulu, and dried inflorescence is used as Mongolian medicine LanCitou. The studies on Chinese medicine Yuzhou Loulu and Mongolian medicine LanCitou in recent years were systemized and compared with each other in the paper, including chemical composi-tion, process research, quality control, pharmacological effects and so on. The results can provide scientific basis for the further studies and utilization of Echinops latifolius Tausch.
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<p><b>OBJECTIVE</b>To investigate the chemical constituents of the flowers of Chrysanthemum indicum.</p><p><b>METHOD</b>The chemical constituents were isolated by various column chromatographic methods. The structures were identified by spectral data.</p><p><b>RESULT</b>Twelve compounds were isolated and identified as acacetin (1), tricin (2), 2',4'-dihydroxychalcone(3), 5-hydroxy-4',7-dimethoxyflavon(4),7hydroxyflavonone (5), isorhamnetin (6),5,6,7-trihydroxy- 3',4', 5'-trimethoxyflanon (7 ), quercetin (8) , (3 beta, 5 alpha, 6 beta, 7 beta, 14 beta)-eudesmen-3,5,6,11-tetrol (9), syringaresinol (10), liriodendrin (11), and genkwanin (12).</p><p><b>CONCLUSION</b>Compounds 3-7, 10-12 were isolated from this species for the first time, and compounds 3, 5, 7, 10, 11 were obtained from genus Chrysanthemum for the first time.</p>
Subject(s)
Chalcones , Chrysanthemum , Chemistry , Flavones , Flavonoids , Flavonols , Flowers , Chemistry , Furans , Glucosides , Lignans , QuercetinABSTRACT
<p><b>OBJECTIVE</b>To investigate the chemical constituents in the leaves and branches of Myricaria alopecuroides.</p><p><b>METHOD</b>Solvent extraction method was employed to extract and partition. The chemical constituents were isolated by column chromatography on silica gel, Sephadex LH-20, highly porous resin HP-20. The structures of the compounds were elucidated on the basis of physiochemical properties and spectral analysis.</p><p><b>RESULT</b>Eleven compounds were isolated from this plant and identified as ellagic acid 3,3',4-trimethylether (1), ellagic acid 3,3'-dimethylether (2), isorhamnetin (3), kaempferol (4), 3, 5-dihydroxy-4-methoxybenzoic acid (5), daucosterol (6), 6,7,10-trihydroxy-8-octadecenoic acid (7), quercetin (8), gallic acid (9), palmitic acid (10), hexadecanoic acid, 2,3-dihydroxypropyl ester (11).</p><p><b>CONCLUSION</b>Except 8 and 9, all compounds were isolated from M. alopecuroides for the first time. Compound 1, 2, 5, 7, 10, 11 were obtained from the genus Myricaria for the frist time.</p>
Subject(s)
Organic Chemicals , Plant Leaves , Chemistry , Plant Stems , Chemistry , Tamaricaceae , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the chemical constituents of the leaves of Aquilaria sinensis, and provide a certain of basis for the comprehensive uses of the plant of A. sinensis.</p><p><b>METHOD</b>The chemical constituents were isolated by various column chromatographic method. The structures were identified by spectral analyses of NMR, MS, et al.</p><p><b>RESULT</b>Thirteen compounds were isolated and identified as 7-hyroxy-5, 4'-dimethoxy flovone (1), 5-hydroxy-7, 4'-dimethoxy flavone (2), luteolin-7-3',4'-trimethyl (3), isocorydine (4), 4-hydroxybenzoic acid (5), triacontenoic (6), hentriacontane (7), alpha-stigmasterol (8), epifriedelanol (9), friedelan (10), friedelin (11), genkwanin (12), 5, 4'-dihyroxy-7, 3'-dimethoxy flovone (13).</p><p><b>CONCLUSION</b>Compound 4 was obtained from this genus for the first time, compounds 1, 6-11, 13 were obtained from this species for the first time.</p>
Subject(s)
Magnetic Resonance Spectroscopy , Organic Chemicals , Chemistry , Plant Leaves , Chemistry , Thymelaeaceae , ChemistryABSTRACT
Objective: To study the chemical constituents of Potentilla multifida L. Methods: Chemical constituents were isolated by the repeated silica gel chromatography and Sephadex LH 20, and their structures were identified by the spectral analysis. Results: Five compounds were obtained as follows: 3?,24 dihydroxyl urs 12 ene (1), ursolic acid (2), euscaphic acid (3), tormentic acid (4), and epihedaragenin (5). Conclusion: Five compounds were isolated from this plant for the first time. Compounds 1 and 5 were isolated from this genus for the first time. Compound 1 was a new natural product.