Astragalin induces autophagy of astrocytes in anterior cingulate cortex of inflammatory pain mice / 中国病理生理杂志

AIM:To investigate the effects of astragalin(AST)on activation status of astrocytes and the ex-pression level of autophagy-related proteins in the cortex of the anterior cingulate cortex of mice with a complete Freund's adjuvant(CFA)-induced inflammatory pain model.METHODS:Twenty-four 6-month-old male C57BL/6 mice were ran-domly divided into four groups:control group,saline group,CFA model group and CFA+60 mg/kg AST administration group,and six mice in each group.Mice in the AST administration group received 60 mg/kg AST by intraperitoneal injec-tion on a body weight basis for 21 d.The paw withdrawal threshold in each group of mice was evaluated by the von Frey test.The expression levels of autophagy-related factors LC3,p62,ATG12 and beclin-1,and astrocyte activation were de-tected by multiplex immunofluorescence staining in the anterior cingulate cortex of mice in each group.Western blot was used to measure the levels of autophagy-related proteins LC3,p62,ATG12 and beclin-1 in the anterior cingulate cortex of mice in each group.RESULTS:Behavioural tests showed that AST significantly increased mechanical pain thresholds in CFA mice(P＜0.05).The results from multiple immunofluorescent staining showed that AST significantly increased the fluorescence intensity of LC3(P＜0.01),ATG12(P＜0.01)and beclin-1(P＜0.05),attenuated the fluorescence intensi-ty of p62(P＜0.05),and inhibited the activation of astrocytes in the anterior cingulate cortex of CFA mice.Western blot results further confirmed that AST significantly increased the expressions of LC3(P＜0.01),ATG12(P＜0.01),beclin-1(P＜0.01),and decreased the expression of p62(P＜0.05)in the anterior cingulate cortex of CFA mice.CONCLU-SION:AST relieves CFA-induced inflammatory pain of mice,and its analgesic mechanism may be related to the inhibi-tion of activation of cortical astrocytes in the anterior cingulate cortex and the promotion of autophagy in CFA mice.

Astragalin Inhibits Apoptosis of Cerebral Cortex Neurons in APP/PS1 Mice / 中山大学学报(医学科学版)

ObjectiveTo investigate the effect of Astragalin （AST） on apoptosis of cerebral cortex neurons in APP/PS1 transgenic mice. MethodsEighteen six-month-old male APP/PS1 transgenic mice were randomly divided into APP/PS1 group， APP/PS1+ 40 mg/kg AST group and APP/PS1+ 20 mg/kg Donepezil （DNP） group， with six mice in each group. At the same time， six male C57BL/6 mice were selected as the normal control group. After intraperitoneal injection of AST once a day and continuous administration for one month， we used Tunel staining to detect the apoptosis of neurons in the cerebral cortex of APP/PS1 mice； immunofluorescent staining to examine the expression of apoptosis-related proteins Bax， Bcl-2， Caspase9 and Cleaved-Caspase3 in the cerebral cortex neurons of APP/PS1 mice； Western blot method to evaluate the changes of the expression of Bax， Bcl-2， Caspase9 and Caspase3. ResultsTunel staining showed that 40 mg/kg AST and 20 mg/kg DNP both reduced the apoptosis of neurons in the cerebral cortex of APP/PS1 mice， AST with more significant inhibition effect. Immunofluorescent staining revealed that 40 mg/kg AST and 20 mg/kg DNP both inhibited the expression of Bax， Caspase9， and Cleaved-Caspase3， and icreased the expression of Bcl-2 in the cerebral cortex neurons of APP/PS1 mice. Western blot results further confirmed that 40 mg/kg AST and 20 mg/kg DNP both down-regulated the expression of Bax （P < 0.05， P < 0.05）， Caspase9 （P < 0.005， P < 0.05） and Caspase3 （P < 0.0001， P < 0.0001） ， and up-regulated the expresstion of Bcl-2 （P < 0.05， P < 0.05） in the cerebral cortex neurons of APP/PS1 mice. ConclusionsAST can inhibit the apoptosis of cerebral cortex neurons in APP/PS1 mice.

Effects of astragalin on the cell proliferation and cell cycle of prostate cancer cells through up-regulating miRNA-513 expression / 肿瘤研究与临床

Objective:To investigate the effects of astragalin on the cell proliferation and cell cycle of prostate cancer cell line C4-2B through up-regulating the expression of miRNA-513 (miR-513).Methods:Prostate cancer cell line C4-2B cells were taken and treated with 125 μg/L of astragalin for 48 h (astragalin group), and untreated C4-2B cells were set as the control group. The methyl thiazolyl tetrazolium (MTT) method was used to detect the proliferation ability of C4-2B cells in the two groups, and cell cycle was detected by using flow cytometry. The miRNAMap prediction software was used to predict that the targeted gene of miR-513 was the forkhead box protein R2 (FOXR2), and the dual luciferase gene reporter assay was used to verify it. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-513 and FOXR2 mRNA in the two groups of cells. Western blotting was used to detect the expressions of FOXR2, cyclin-dependent kinase 7 (CDK7), β-actin and cyclin H in the two groups of C4-2B cells.Results:Compared with the control group, the proliferation activity of C4-2B cells in the astragalin group was decreased from day 2 to day 5 (all P < 0.05). The proportions of S-phase cells in the control group and the astragalin group were (48.1±3.2)% and (36.0±2.1)%, respectively. The proportion of S-phase cells in the astragalin group was decreased ( t = 3.12, P = 0.021); the proportions of G 2-phase cells were (24.9±3.3)% and (11.8±2.4)%, respectively. The proportion of G 2-phase cells in the astragalin group was decreased ( t = 3.18, P = 0.019). The relative expression levels of miR-513 in C4-2B cells of the control group and the astragalin group were 1.01±0.22 and 6.55±0.61, respectively. The relative expression levels of miR-513 in C4-2B cells in the astragalin group was increased ( t = 7.70, P < 0.01). The dual luciferase reporter gene assay verified that FOXR2 was the targeted gene of miR-513. The relative expression level of FOXR2 mRNA in C4-2B cells of the control group and the astragalin group was 1.04±0.14 and 0.19±0.06, respectively, and the difference was statistically significant ( t = 5.53, P = 0.002), suggesting that after astragalin promoted the expression of miR-513, the FOXR2 mRNA expression was decreased. The relative expression levels of FOXR2, CDK7 and cyclin H protein in C4-2B cells in the astragalin group were all decreased compared with those in the control group. Conclusions:Astragalin inhibits the proliferation of prostate cancer C4-2B cells and induces cell cycle arrest by up-regulating the expression of miR-513.

Effect of astragalin on apoptosis of human colon cancer ISHIKAWA cells / 中华内分泌外科杂志

Objective:To investigate the apoptosis-inducing effect of baohuoside I (BI) on endometrial cancer Ishikawa cells and its related molecular mechanism.Methods:With 0 μ M and 0 h treatment were used as blank control group, and BI treatment was used as experimental group. The inhibitory effect of BI on the proliferation of Ishikawa cells was detected by CCK-8 assay. The apoptosis-inducing effect of BI on Ishikawa cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The expressions of apoptosis-related proteins and signaling pathway-related proteins were detected by Western blot.Results:CCK-8 experiment showed that BI could be expressed in concentration gradient (3, 10, 20, 30, 40 μM). It could effectively inhibit the proliferation of Ishikawa cells (the survival rates were 89.56±0.96, 74.69±1.21, 60.28±1.09 and 43.51±2.17 respectively). Its toxic and side effects on normal cells were lower than that of 5-FU. The results of flow cytometry showed that BI could effectively induce the apoptosis of Ishikawa cells by reducing the level of mitochondrial membrane potential. The proportion of apoptotic cells in each group was (9.92±0.77) %, (14.01±0.83) %, (17.05±1.41) %, (28.21±1.73) % and (44.55±3.11) %. Western blot showed that BI could up-regulate the level of p-p38 and reduce the level of p-STAT3.Conclusions:BI can effectively inhibit the proliferation of Ishikawa cells, and induce apoptosis by reducing the mitochondrial membrane potential and activating the mitochondria-dependent pathway. Its regulatory mechanism is achieved by activating the p38 signaling pathway and inhibiting the STAT3 pathway.

Simultaneous Determination of 7 Active Components in Mori Australis Cortex and Mori Cortex from Different Sources by HPLC / 中国药房

OBJECTIVE：To establish a meth od for the simultaneous determination of 7 active components in Mori Australis Cortex and Mori Cortex from different sources in Chongqing area ，so as to provide reference for improving the quality control standards of Mori Australis Cortex and Mori Cortex and comparing the equivalence of their quality. METHODS ：HPLC method was used to determine the contents of neochlorogenic acid ，mulberroside A ，chlorogenic acid ，astragalin，kaempferol，morusin and isoquercetin in 58 batches of Mori Australis Cortex and Mori Cortex. The chromatographic column was Diamonsil C 18 with mobile phase consisted of 0.1％ formic acid solution-acetonitrile （gradient elution ） at the flow rate of 1.0 mL/min. The detection wavelength was 280 nm，column temperature was 30 ℃，and the injection volume was 10 μL. Using SPSS 22.0 software， independent sample t-test，principal component analysis and cluster analysis were used to analyze the content difference of the above-mentioned 7 active components in Mori Australis Cortex and Mori Cortex. RESULTS ：There was a good linear relationship between the peak area and the concentration of the above 7 active components （r≥0.999 0）. The RSDs of precision ，stability（24 h），repeatability，durability and recovery were less than 3％. The average contents of neochlorogenic acid ，mulberroside A ， chlorogenic acid ， astragalin， kaempferol， morusin and 023-58576130。E-mail：1025473978@qq.com isoquercetin in Mori Australis Cortex were 0.304，22.462， 1.730，1.308，1.593，2.842 and 0.657 mg/g，respectively. Those of Mori Cortex were 0.305，22.995，2.486，2.438， 2.916，4.158 and 1.264 mg/g，respectively. The results of independent sample t-test showed that only the content of kaempferol in the above 7 active components of Mori Australis Cortex and Mori Cortex had significant difference （P＜0.05）. The results of principal component analysis and cluster analysis showed that there was no significant difference in the contents of above 7 active components between Mori Australis Cortex and Mori Cortex. CONCLUSIONS：The established HPLC method is simple ，sensitive and accurate ，which can provide a reference for improving the quality control standard of Mori Australis Cortex and Mori Cortex. Mori Australis Cortex and Mori Cortex have certain quality equivalence in main active components ，and the Mori Australis Cortex from M. australis and M. cathayana can be used as a substitute for the Mori Cortex.

Astragalin inhibits airway inflammation in asthma through NOX2/ROS/NF-κB signaling pathway / 中国药理学通报

Aim To investigate the effect of astragalin (AG) on airway inflammation in asthmatic mice and its mechanism. Methods Fifty SPF male mice were randomly divided into normal group, asthma model group, and astragalin low (AG25), medium (AG50), and high (AG100) dose groups. A mouse model of asthma was prepared by egg albumin inhalation, the number of cells was counted by Diff-Quik staining after collecting bronchoalveolar lavage fluid (BALF), and IL4, 5, and 13 levels in BALF were measured by ELISA. The inflammatory changes in mouse lung tissues were observed using HE staining. Reactive oxygen species (ROS) levels were measured using a DCFH-DA probe, and superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured by colorimetry. IL-4, IL-5, IL-13, NOX2, p47phox, p-NF-κBp65, NF-κBp65, IκBα, p-IκBα and β-actin expressions in lung tissues were detected by Western blot. Results AG significantly reduced the number of inflammatory cells and total cells in BALF, decreased IL-4, IL-5, and IL-13 contents in BALF and lung tissues, and reduced inflammatory cell infiltration in lung tissues. AG inhibited nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) and p47phox expression, decreased ROS levels and MDA levels, increased SOD activity, and inhibited IκBα and NF-κBp65 phosphorylation. Conclusion AG attenuates airway inflammation in asthma by modulating the oxidative stress response through the NOX2/ROS/NF-κB signaling pathway.

Screening and determination of estrogen-like quality markers of Cuscuta chinensis / 中草药

Objective: To preliminarily screen out the estrogen-like quality markers of small grain Cuscuta chinensis from Heilongjiang Province, so as to provide reference for its subsequent experimental research and quality control. Methods: UPLC- Q-TOFMS/MS was used to qualitatively analyze the extract of C. chinensis and the fingerprints of different polar fractions were established. The estrogenic activity of different polar fractions was evaluated with uterine coefficient, endometrial thickness and serum estrogen level of mice. The bivariate correlation analysis and gray relational analysis were used to construct the composition-activity relationship between chemicals and the effects of estrogen for screening quality markers. And the content determination methods of the five quality markers were established. Results: A total of 10 quality markers related to the estrogenic effect from C. chinensis were found in the positive and negative ion scanning modes. They were hyperoside, astragalin, stigmasterol, neocuscutoside C, apigenin, kaempferol, 6-O-(E)-p-coumaroyl-β-D-fructofuranosyl-(2→1)-α-D-glucopyranoside, quercetin, isorhamnetin, and 2,6-octadecadiynoic acid. The contents of the five quality markers were determined as follows: hyperoside (2.753 ± 0.097) mg/g, quercetin (1.139 ± 0.107) mg/g, apigenin (1.104 ± 0.047) mg/g, kaempferol (1.144 ± 0.079) mg/g and isorhamnetin (0.697 ± 0.074) mg/g. Conclusion: The quality markers of small grain C. chinensis from Heilongjiang Province can be screened out according to the composition-activity relationship, and the method for the detect the concentrations of the quality markers is accurate and stable.

Chemical constitutents from flowers of Stellera chamaejasme and their antioxidant activity / 中草药

Objective: To investigate the flavonoids and lignans from the flowers of Stellera chamaejasme and their structure-activity relationship (SAR) of antioxidant activity. Methods: The compounds were isolated by column chromatography and HPLC packed with macroporous resin, silica gel, and Sephadex LH-20. Their structures were elucidated by spectroscopic analysis. Their anti-oxidant activities in vitro were evaluated by DPPH, ABTS, and FRAP assays. Results: Twelve compounds were isolated from the flowers of S. chamaejasme, and identified as artemisetin (1), quercetin (2), isoscutellarein-8-O-β-D-glucuronopyranoside (3), quercetin-3-O-β-D- glucopyranoside (4), astragalin (5), hypolaetin-8-O-β-D-glucuronopyranoside (6), kaempferol 3-O-β-D-glucopyranosyl-(1→2)-O-α- L-xylopyranoside (7), rel-(3R,3'S,4R,4'S)-3,3',4,4'-tetrahydro-6,6'-dimethoxy [3,3'-bi-2H-benzopyran]-4,4'-diol (8), matairesinol (9), uralenol (10), cycloastragenol (11), and (+)-pinoresinol (12). Conclusion: Compounds 1, 3, 5-7, and 10 are isolated from this plant for the first time, and compounds 2, 4, 5, and 10 showed significant antioxidant activity, and the SAR analysis suggested that the glycosylation at the C-8 or C-3 position of flavonoids could weaken their antioxidant activity.

Comparative study on multiple chemical pattern recognition combined with fingerprint of Tetrastigma hemsleyanum from different habitats / 中草药

Objective: To compare the identification effects of different pattern recognition methods combined with chemical fingerprints on Tetrastigma hemsleyanum from different habitats, and propose a new identification method for the habitats of T. hemsleyanum. Methods: A total of 72 batches of T. hemsleyanum samples were collected from Zhejiang, Yunnan and Guizhou. HPLC fingerprints were collected, 18 common peaks were marked, and the difference of principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA) and random forest (RF) in processing complex data of samples from different habitats was compared. Results: A total of 72 batches of T. hemsleyanum samples can only be divided into two categories by PCA, the results of OPLS were better than PCA, and the RF can completely separate the samples from three habitats. The RF combined with fingerprint can effectively identify and distinguish T. hemsleyanum from different habitats. Conclusion: This study can be used as an effective method for the quality control of T. hemsleyanum from different habitats and provide an effective reference for multi-index complex fingerprint identification from different habitats.

Protective effect of Rhododendron weyrichii flower extract against UVB-induced proinflammatory cytokine production in human keratinocytes

Ultraviolet B (UVB) radiation is harmful to the skin and induces cytokine release from keratinocytes leading toinflammatory skin disorders. Previous studies have shown that chronic exposure to UVB radiation increases tumornecrosis factor (TNF)-α and interleukin (IL)-6 secretion through various signaling pathways, resulting in skininflammation and increased risk of skin cancer. The present study was undertaken to investigate the protective effectsof Rhododendron weyrichii flower (RWF) extracts against UVB damage of immortalized human keratinocytes(HaCaT). To determine the anti-inflammatory effects of RWF, we examined UVB-induced proinflammatory cytokineproduction in HaCaT cells in the presence or absence of RWF extract, using enzyme-linked immunosorbent assay(ELISA). The results indicated that the RWF extract inhibited the production of proinflammatory molecules suchas IL-6 and TNF-α, but not IL-8, in UVB-irradiated HaCaT cells. These results demonstrate that RWF potentiallyprotects against UVB-induced skin inflammation. In addition, using high-performance liquid chromatography (HPLC)fingerprinting, kaempferol (0.335 ppm) and astragalin (2.569 ppm) were identified and quantified as RWF extractconstituents. Moreover, we tested the potential application of RWF extracts as a cosmetic treatment by performinghuman skin primary irritation tests. In these tests, the RWF extracts did not induce adverse reactions. Based on theseresults, we suggest that RWF extracts be considered anti-inflammatory candidates for pharmaceutical and/or cosmeticapplications.

Determination of 25 characteristic components in Danhe Granules by LC-MS and consistency analysis of preparations / 中草药

Objective: A high performance liquid chromatography coupled to triple quadrupole mass spectrometry method (HPLC- MS/MS) was established to simultaneously determine the content of 25 characteristic components (gallic acid, tanshinol, catechin, chlorogenic acid, caffeic acid, epicatechin, rutin, polydatin, hyperin, astragalin, naringin, hesperidin, rosmarinic acid, resveratrol, salvianolic acid B, quercetin, emodin-8-O-β-D-glucoside, isorhamnetin, emodin, coclaurine, nuciferine, cryptotanshinone, tanshinone I, dehydronuciferine, tanshinone IIA) in Danhe Granules (DG), and the consistency between different batches was investigated. Methods The analysis was conducted on Agilent Rapid Resolution HD C18 column (50 mm × 2.1 mm, 1.8 μm) with the mobile phase of 0.1% formic acid water-0.1% formic acid acetonitrile. The dynamic multi-response detection (dMRM) scanning mode was used in the mass spectrometry. Results: Based on the established HPLC-MS/MS method, the simultaneous quantitative analysis of 25 characteristic components could be completed within 10 min, with the quantitative limits of isorhamnetin, coclaurine, and dehydronuciferine of 0.025 ng/mL; emodin and tanshinone I of 0.050 ng/mL; emodin-8-O-β-D-glucoside of 0.200 ng/ml; gallic acid, chlorogenic acid, caffeic acid, epicatechin, rutin, hyperin, astragalin, resveratrol, quercetin, nuciferine, tanshinone IIA of 0.250 ng/ml; catechin, rosmarinic acid, and cryptotanshinone of 0.500 ng/mL; tanshinol, hesperidin, and salvianolic acid B of 1.000 ng/mL; polydatin of 2.000 ng/mL; naringin of 5.000 ng/mL, respectively. The linear relationships of the 25 constituents within their respective mass concentrations were good, with the average recovery of 85.16%-113.46% and the RSD of 2.01%-8.80%. Furthermore, this method also included the main components named monarch, minister, assistant and guide of herbs in a relatively comprehensive way. The total content of the 25 components was 31.49 mg/g, among which the content of salvianolic acid B (9.44 mg/g) and hesperidin (7.60 mg/g) was the highest, and the content of isorhamnetin (0.79 μg/g) was the lowest. According to boxplot analysis, the content of 25 components in 10 different batches of DG fluctuated (P value) within 75% < P < 125%; and the RSD value of 25 components ranged from 2.58% to 13.10% by statistical analysis. The above results showed that the consistency of the component content among 10 batches of DG was acceptable. Conclusion: The analytical method established in this study is fast and sensitive. Furthermore, the results of this study are reliable and can provide scientific methods and basis for quality control and consistency analysis of DG.

Simultaneous Determination of Isoquercitrin ，Astragalin and Salvianolic Acid B in Moringa oleifera Leaves Granules by HPLC / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of isoquercitrin， astragalin and salvianolic acid B in Moringa oleifera leaves granules. METHODS： HPLC method was adopted. The determination was performed on Cosmosil-C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at flow rate of 1.3 mL/min. The column temperature was 40 ℃ and detection wavelength was set at 260 nm. The sample size was 10 μL. RESULTS： The linear ranges of isoquercitrin， astragalin and salvianolic acid B were 0.017-0.341， 0.010-0.194， 0.010-0.195 mg/mL， respectively （all r＞0.999 0）. The detection limits were 0.085， 0.143， 0.117 μg/mL， and the limits of quantitation were 0.283， 0.476， 0.392 μg/mL， respectively. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2.0％ （n＝6）. Average recoveries were 101.22％， 98.76％ and 98.72％， and RSDs were 0.66％，0.30％，0.30％ （n＝6）， respectively. CONCLUSIONS： The established method is simple， accurate and reproducible. It can be used for simultaneous determination of isoquercitrin， astragalin and salvianolic acid B in M. oleifera leaves granules.

Study on HPLC characteristic spectra of Lygodii Herba and chemical compositions of different parts from Lygodii Herba / 中草药

Objective: To establish the HPLC characteristic spectrum of Lygodii Herba from different habitats in Guangxi Province, and to evaluate the difference of components in different parts of medicinal materials. Methods: HPLC was performed on Waters symmetry C18 (250 mm × 4.6 mm, 5 μm) column, with the mobile phase of acetonitrile-0.2% phosphoric acid at flow rate of 1.0 mL/min; Detection wavelength was 354 nm; The column temperature was 30 ℃ and the sample size was 20 μL. Twelve batches of Lygodii Herba samples were determined and the characteristic spectrum of those were established, and the content of rutin, isoquercetin, and astragalin were determined to evaluate the difference of chemical components in different parts of medicinal materials. Results: There were seven characteristic peaks identified in the characteristic spectra of Lygodii Herba samples. Peak 1 was caffeic acid, peak 2 was Rutin, peak 3 was Isoquercitrin and peak 5 was astragalin. The similarities of 11 batches of samples were proved to be higher than 0.900 and one batch of them was proved to be less than 0.900. The chemical component of stem was richer than that of leaves of Lygodii Herba, and the content of the component was higher in the stem than that of the leaves. Conclusion: The method is simple, accurate, and reproducible, which can provide the scientific evidence for controlling the internal quality standards effectively. The preferred harvest season of Lygodii Herba is autumn and winter.

Optimization of extraction and purification technology for phenolic acids and flavonoids in stems and leaves of Salvia miltiorrhiza / 中草药

Objective To study the optimum condition for the extraction and purification of phenolic acids and flavonoids in the stems and leaves of Salvia miltiorrhiza. Methods The contents of phenolic acids and flavonoids were determined by ultra performance liquid chromatography (UPLC). The extraction process was evaluated by single factor test and orthogonal design with yield of dry extract and the content of phenolic acids (including danshensu, protocatechuic acid, protocatechualdehyde, caffeic acid, salvianolic acid B, rosmarinic acid, and lithospermic acid) and flavonoids (including rutin, isoquercitrin, kaempferol-3-O-rutinoside, and astragalin) as index. The effects of extraction method, extraction solvent, ratio of material to liquid, extracting time, and extracting times on the extraction of stems and leaves of S. miltiorrhiza were investigated. Macroporous adsorption resin was used to purify the sample, and the purification process parameters were investigated to determine the optimum purification process. Results The optimum condition for the extraction of the stems and leaves of S. miltiorrhiza is that eight times of 50% ethanol for three times reflux extraction and 1 h for each time and AB-8 macroreticular resin was selected for the purification. Optimum process was as following:The concentration of sample solution was 1.0 g/mL; The loading quantity of the sample was 0.15 g dried extract of per gram; The resin column chromatography was eluted with 3 BV of 40% ethanol. Under these conditions, the total purity of phenolic acids and flavonoids could reach 40.83%. Conclusion The optimum technology is stable and feasible for the extraction and purification of phenolic acids and flavonoids in the stems and leaves of S. miltiorrhiza, and can provide reference for further development and utilization.

Determination of seven effective constituents in crude and processed Cuscuta australis by quantitative analysis of multi-components with single marker / 中草药

Objective To establish a quantitative analysis of multi-components with single marker (QAMS) for the simultaneous determination of chlorogenic acid, cryptochlorogenin acid, caffeic acid, hyperoside, isoquercitrin, astragalin, kaempferol in crude and processed Cuscuta australis, which is proved to be a scientific and feasible method in the quality analysis in C. australis. Methods Six relative correction factors (RCFs) of chlorogenic acid, cryptochlorogenin acid, caffeic acid, isoquercitrin, astragalin, kaempferol was established in the HPLC method with the hyperoside as the internal standard (IS), which was to calculate the mass fraction of each. The mass fraction of seven effective constituents in crude and processed C. australis was calculated by the external standard method (ESM) at the same time. Compared with the content results determined by the ESM and QAMS, the feasibility and accuracy of QAMS method were verified. Results The relative correction factor (RCF) was perfect. The detection calculated by QAMS was consistent with the results by ESM. Conclusion The method with a single marker, using the hyperoside as IS, is accurate and feasible for the quantitative analysis of six other effective constituents in C. australis.

Content Determination of 6 Active Components in Morus alba by HPLC / 中国药房

OBJECTIVE:To establish the method for the simultaneous determination of content of 6 active components as neochlorogenic acid,mulberroside A,chlorogenic acid,astragalin,sanggenon C and morusin in Morus alba,and to provide reference for improving quality control standard of M. alba. METHODS:HPLC method was adopted. The determination was performed on Agilent 5 TC-C18with mobile phase consisted of acetonitrile-0.1% formic acid(gradient elution)at the flow rate of 1 mL/min. The detection wavelength of 280 nm. RESULTS:The mass concentration linear range of neochlorogenic acid, mulberroside A,chlorogenic acid,astragalin,sanggenon C and morusin were 0.001 06-0.042 4,0.001 67-0.066 8,0.007 95-0.318, 0.001 65-0.066 0,0.005 00-0.200 and 0.001 24-0.049 6 mg/mL,respectively(all r≥0.999 6);the limits of quantitation were 0.11, 0.14,0.81,0.17,0.45 and 0.12 μg/mL,respectively;the limits of detection were 0.04,0.05,0.41,0.07,0.18 and 0.04 μg/mL, respectively;RSDs of precision test were 0.26%,0.31%,0.24%,0.27%,0.36% and 0.44%(n=6),respectively;RSDs of stability test were 0.68%,0.54%,0.62%,0.53%,0.41% and 0.73%(n=6),respectively;average method recovery rates were 99.1%,98.8%,98.8%,98.4%,98.5% and 99.9%(RSDs were 0.5%-1.5%,n=9),respectively. CONCLUSIONS:The method is simple,accurate,and can be used for simultaneous determination of 6 active components in M.alba.

In silico pharmacodynamics, toxicity profile and biological activities of the Saharan medicinal plant Limoniastrum feei

ABSTRACT In-silico study was performed to find the pharmacodynamics, toxicity profiles and biological activities of three phytochemicals isolated from Limoniastrum feei (Plumbagenaceae). Online pharmacokinetic tools were used to estimate the potential of Quercetin, kaempferol-3-O-ß-D-glucopyranoside (astragalin) and quercitin-7-O-ß-D-glucopyranoside as specific drugs. Then the prediction of potential targets of these compounds were investigated using PharmMapper. Auto-Dock 4.0 software was used to investigate the different interactions of these compounds with the targets predicted earlier. The permeability of quercetin was found within the range stated by Lipinski ׳s rule of five. Hematopoietic prostaglandin (PG) D synthase (HPGDS), farnesyl diphosphate synthetase (FPPS) and the deoxycytidine kinase (DCK) were potential targets for quercetin, astragalin and quercetin 7, respectively. Quercetin showed antiallergic and anti-inflammatory activity, while astragalin and quercetin 7 were predicted to have anticancer activities. The activity of Astragalin appeared to be mediated by FPPS inhibition. The inhibition of DCK was predicted as the anticancer mechanisms of quercetin 7. The compounds showed interesting interactions and satisfactory binding energies when docked into their targets. These compounds are proposed to have activities against a variety of human aliments such as allergy, tumors, muscular dystrophy, and diabetic cataracts.

Chemical constituents of Swertia bimaculataa / 中草药

Objective: To investigate the chemical constituents from the dried whole plants of Swertia bimaculata. Methods: The compounds were isolated and purified by various column chromatography, and their structures were elucidated on the basis of chemical and spectroscopic analyses. Results: Thirty compounds were isolated from the n-BuOH fraction of S. bimaculata EtOH extract and identified as 1,3,6-trihydroxy-4,7-dimethoxyxanthone (1), 1,8-dihydroxy-2,3,4,5-tetramethoxyxanthone (2), 1,3,5-trihydroxy-4- methoxyxanthone (3), 1,3-dihydroxy-5-methoxyxanthone (4), 1,3,5-trihydroxyxanthone (5), 1,3,7-trihydroxy-4-methoxyxanthone (6), 2-(3'-O-β-D-glucopyranosyl) benzoyloxygentisic acid (7), veratrilogenin (8), demethylbellidifolin (9), isobellidifolin (10), 1,3,8-trihydroxy-4,5-dimethoxyxanthone (11), 1,3-dihydroxy-4,5-dimethoxyxanthone (12), methylbellidifolin (13), 1-hydroxy-2,3,4,7-tetramethoxyxanthone (14), mangiferin (15), astragalin (16), isoquercitrin (17), nicotiflorin (18), rutin (19), isoorientin-7,3'-dimethylether (20), swertisin (21), swertiajaponin (22), isoorientin (23), isoscutellarein-8-methyl ether (24), 8-chrysoeriol (25), kaempferol (26), luteolin (27), quercetin (28), β-sitosterol (29), and oleanolic acid (30). Conclusion: Compound 1 is a new compound. named bimaculatone A. Compounds 2-7, 16-20, and 24-26 are firstly obtained from the plants of Swertia L., and compounds 2-11 and 16-28 are obtained from S. bimaculata for the first time.

Effects of astragalin on pulmonary edema in acute cervical spinal cord injury rats / 中华创伤杂志

ObjectiveTo investigate the effects of astragalin on pulmonary edema after acute cervical spinal cord injury in rats.MethodsA total of 200 adult Wistar rats weighing 240-250 g were randomly divided into five groups:astragalin group,low concentration astragalin group,physiological saline group,blank group and sham group,with 40 rats in each group.The rats with cervical spinal cord injury were induced at C7 by modified Allen' s method,with the dropping weight of 10 × 2.5 g · cm.In the sham group,the laminas were removed only,leaving spinal cord at the C7 intact.Each group was further divided into four time points:24 hours,3 days,1 week and 2 weeks after the modeling,with 10 rats in each time point,according to the specimen collection time.Rats were sacrificed at different time points to observe the pathological change of the lung tissue using optical microscope,measure the lung wet/dry weight ratio (W/D) and protein concentrations of the serum and bronchoalveolar lavage fluid and calculate the lung W/D and lung permeability index (LPI).ResultsAt the same instant,the W/D and LPI in the astragalin group and low concentration astragalin group were lower than those in physiological saline group and blank group,with the lowest value in the astragalin group at day 3 after injury ( P ＜0.05 ).ConclusionsRats with acute cervical spinal cord injury may cause pulmonary edema,which can be efficiently alleviated through early use of astragalin.

Chemical constituents in roots and rhizomes of Trillium tschonoskii / 中草药

Objective: To investigate the chemical constituents in the roots and rhizomes of Trillium tschonoskii. Methods: The roots and rhizomes of T. tschonoskii were extracted with 70% ethanol and separated by polyamide, silica gel, RP-C18, and Sephadex LH-20 column chromatography. Chemical structures were identified by MS, 1D and 2D NMR spectroscopy. Results: Eight compounds were isolated and their structure were identified as pennogenin-3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside (1), pennogenin-3-O-α-L-rhamno-pyranosyl-(1→4)-[α-L- rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (2), pennogenin-3-O- α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4) -[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (3), pennogenin (4), prosapogenin (5), 7, 11-dimethyl-3-methylene-1, 6-dodecadien-10, 11-dihdroxyl-10-O-β-D-glucopyranosyl-(1→4)-O-β-D-glucopyranoside (6), 7, 11-dimethyl-3-methylene-1, 6-dodecadien-10, 11-dihdroxyl-10-O-β-D- glucopyranoside (7), and astragalin (8). Conclusion: All these compounds are isolated from T. tschonoskii for the first time.