Water extract of Glycyrrhiza uralensis Fisch.enhanced the activation of RAW264.7 cells through TLR4 signaling pathway / 中华微生物学和免疫学杂志

Objective To investigate the effects of water extract of Glycyrrhiza uralensis Fisch.(GUWE) on the activation of RAW264.7 cells and the possible mechanism.Methods RAW264.7 cells were treated with GUWE containing different concentrations of polysaccharide (10, 50, 100, 500 μg/ml).Viability of these cells was analyzed by MTT assay.Phagocytic activity and surface molecules expressed on these cells were detected by flow cytometry.Levels of cytokines were analyzed by ELISA.Western blot assay was performed to analyze the activation of key molecules in TLR4 signaling pathway.Results GUWE at the concentration of 500 μg/ml significantly decreased the viability of RAW264.7 cells, but significantly increased the viability of RAW264.7 cells at concentrations of 50 μg/ml and 100 μg/ml.GUWE significantly enhanced the phagocytic activity of RAW264.7 cells as well as the expression of cytokines and costimulatory molecules in a dose-dependent manner.Further analysis indicated that the activation of RAW264.7 cells induced by GUWE was suppressed by TLR4 inhibitor.Moreover, GUWE enhanced the phosphorylation of NF-kB p65 and TLR4 downstream mitogen-activated protein kinases (MAPKs) including p38, extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK).Conclusion This study indicates that GUWE promotes the activation of RAW264.7 cells through TLR4 signaling pathway.

Contents Determination of Nine Components in Guizhi Decoction and Comparison of Different Decoction Methods / 中国药房

OBJECTIVE:To establish a method for the contents determination of 9 components in Guizhi decoction,and com-pare the effects of traditional decoction method and the extracting machine decoction method on these contents in Guizhi decoction. METHODS:HPLC was performed on the column of Agilent Zorbax SB-C18 with mobile phaseof acetonitrile- 0.1% phosphoric ac-id(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 230 nm,254 nm and 280 nm,the column tempera-ture was 25℃,and the injection volume was 10μl. RESULTS:The linear range was 0.410 2-210.0μg/ml for gallic acid(r=0.999 9), 0.994 0-254.5μg/ml for albiflorin(r=0.999 9),1.636 0-1 675.0μg/ml for paeoniflorin(r=0.999 9),0.988 3-506.0μg/ml for liquiri-tin(r=0.999 6),0.987 3-31.59 μg/ml for coumarin(r=0.999 5),0.486 8-124.6 μg/ml for cinnamic acid(r=0.999 5),2.458 0-314.6μg/ml for cinnamaldehyde(r=0.999 5),0.034 3-1.096 μg/ml for 2-methoxy cinnamaldehyde(r=0.999 8),and 1.711 0-219.0 μg/ml for glycyrrdhizic acid (r=0.999 7);RSDs of precision,stability and reproducibility tests were lower than 5%,recoveries were 93.56%-103.19%(RSD=4.00%,n=9)、101.51%-107.32%(RSD=2.21%,n=9)、95.08%-103.76%(RSD=2.87%,n=9)、100.82%-105.73%(RSD=1.85%,n=9)、85.08%-89.12%(RSD=1.40%,n=9)、92.31%-99.12%(RSD=2.71%,n=9)、99.17%-102.32%(RSD=1.24%,n=9)、100.15%-103.98%(RSD=1.18%,n=9)、99.93%-102.61%(RSD=1.03%,n=9). The content of total effective components from the extracting machine decoction method was 4 565μg/g,that from the traditional decoc-tion method was 2 742 μg/g.CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of 9 componentsin Guizhi decoction. The contents of gallic acid,albiflorin and 2-methoxy cinnamaldehyde are first re-ported. The total effective components from the extracting machine decoction method are higher than that from the traditional decoc-tion method.

Research on relationship between commercial specifications of Scutellariae Radix and chemical composition and drug quality / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the chemical differences in 4 commercial specifications of Scutellaria Radix, research the affection of decayed central xylem part on the crude drug's chemical composition and provide scientific data for production, processing, sale and clinical applications of Scutellariae Radix.</p><p><b>METHOD</b>Macroscopical identification method was used for observation of different specifications of Scutellariae Radix, including Qinwang, Tiaoqin both in 1st class and 2nd class and inferior samples. HPLC fingerprint method was used to analyze chemically the decayed central xylem part and non-decayed part as well as complete sample, and the results were described by the relative peak area.</p><p><b>RESULT</b>The morphological characteristics of 4 specifications are greatly different from one another mainly in root diameters, root lengths and the proportions of decayed central xylem part in the root, and so the authors classified Qinwang and Tiaoqin in 1st class as Kuqin for all samples of them which have decayed central xylem; and classified Tiaoqin in 2nd class and the inferior samples as Ziqin, for having little decayed central xylem. The 4 specifications collected from the same producing area have similar HPLC fingerprint profile to one another, while they are different in relative peak area. The peak area ratios of aglycone to their glucuronide (baicalein/baicalin, wogonin/wogonoside, oroxylin A/oroxylin A-7-O-glucuronide) from Kuqin were significantly higher than those of Ziqin. The total area of the peaks in HPLC fingerprint chromatographs of decayed central xylem part were quite lower than that of non-decayed part, whereas peak areas of the characteristic peaks and the 3 peak area ratios of decayed central xylem were significantly higher than those of non-decayed part which could be used as characteristic parameters to distinguish Kuqin and Ziqin.</p><p><b>CONCLUSION</b>Four commercial specifications of Scutellariae Radix can be classified as Kuqin and Ziqin respectively according to morphological characteristics and the proportions of decayed central xylem part in the root. The chemical characteristics of Kuqin and Ziqin are different from each other, so it's worth clarifying the similarities and differences of Kuqin and Ziqin in future. The result in this research can be used as references for identification and quality control of Scutellariae Radix specifications, and investigation on effective components of Kuqin and Ziqin.</p>

Determination of harpagide and harpagoside in Scrophulariae Radix by HPLC-UV / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a method for the determination of harpagide and harpagoside in Scrophulariae Radix (Xuanshen) by HPLC-UV under double wavelength, and to study the changes of these two constituents during processing, and to set the limitation of harpagide and harpagoside contents in crude drug and sliced pieces of Xuanshen.</p><p><b>METHOD</b>The analyses were performed on an Agilent Technologies ZORBAX SB-C18 (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-water (containing 0.03% phosphoric acid) in gradient model. The flow rate was 1.0 mL x min(-1) . The column temperature was 25 degrees C. The UV detector wavelength was set at 210 nm before 13 min and then changed to 280 nm.</p><p><b>RESULT</b>Harpagide and harpagoside were separated well. The linear calibration curves were obtained over of 0.0549 - 1.46 microg for harpagide (r = 0.9999, n =7) ,0.0225 - 0.900 microg for harpagoside (r = 0.9998, n = 9). The recoveries ( +/- RSD)% were 98.1 (+/- 2.4)% for harpagide and 98.8 (+/- 4.3)% for harpagoside. The contents of harpagide were 0. 277% - 0.620%, harpagoside were 0.078% - 0.362% in Xuanshen, and harpagide were 0.276% - 1.059%, harpagoside were 0. 059% - 0.183% in sliced Xuanshen, respectively. After the processing of Scrophulariae Radix, the content of harpagide increases 13.7% - 96.0%, while harpagoside decreases 11.0%-73.9%.</p><p><b>CONCLUSION</b>This method is simple, accurate, and can be used for the quality control of Scrophulariae Radix. We propose that the total content of harpagide and harpagoside in either crude drug or sliced pieces of Scrophulariae Radix should not be less than 0.45%.</p>