Prediction and analysis of quality markers between crude and wine-processed Scutellaria baicalensis based on "components-targets-metabolism" network analysis / 中草药

Objective: To explore the potential Q-markers between crude Scutellaria baicalensis (CSR) and wine-processed S. baicalensis (WSR) based on "components-targets-metabolism" network analysis. Methods: According to the differential components between CSR and WSR, the network relationship of "components-target-metabolomics" was constructed combining network pharmacology and metabolomics. The correlation analysis was then conducted between flavonoids glycosides, and aglycones in S. baicalensis, between differential components and endogenous metabolites to predict the potential quality markers. Results: In this study, combining the results of network pharmacology and metabolomics, baicalin and oroxylin A-7-O-glucuronide were regarded as the quality markers of CSR; Baicalein and wogonin were considered as the quality markers of WSR. Conclusion: It is crucial wine-processed mechanism of S. baicalensis that glutinous rice wine can promote the dissolution and absorption of aglycones. Overall, identification of the differences between Chinese herbal decoction pieces and it processed product, combining analysis of network pharmacology and metabolomics, which provides a demonstration for the investigation of quality markers of Chinese herbal pieces.

Chemical constituents of Artemisia rupestris / 中草药

Objective: To investigate the chemical constituents of the whole plants of Artemisia rupestris. Methods: The chemical constituents were isolated and purified by silica gel, Sephadex LH-20 column chromatography and semi-preparative HPLC. The structures of the isolated compounds were identified by NMR and MS spectroscopic method. Results: Eighteen compounds were isolated and purified, which structures were identified as 2α-(2',4'-hexadiynoyl)-1,6-dioxaspiro [4,5]-deca-3-ene (1), chrysophanol (2), ethyl vanillate (3), methyl linolenate (4), 8,11-octadecadienoic acid methyl ester (5), eicosane coumaric acid ester (6), emodin (7), 2,5-di-tert-butylphenol (8), phellopterin (9), syringic acid (10), 7-methoxycoumarin (11), vanillin (12), imperatorin (13), oroxylin A (14), 5-hydroxy-3,4',6,7-tetramethoxyflavone (15), erucylamide (16), octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propanoate (17) and 3,5,3',4'-tetrahydroxy-6,7-dimethoxyflavone (18). Conclusion: Compounds 1, 4, 11-14 and 18 are isolated from A. rupestris for the first time, and all the other compounds were isolated from the genus Artemisia for the first time.

Simultaneous determination of four components of Shuganning Injection in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study / 中国中药杂志

This study aims to reveal the pharmacokinetics of Shuganning Injection in normal rats. In this experiment,ultra-high performance liquid chromatography-electrospray-tandem mass spectrometry( UPLC-ESI-MS/MS) was used to establish an analytical method for simultaneous determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma. Then,the non-compartmental model( NCA) in Phoenix WinN onL in 6. 4 software was used to fit pharmacokinetic parameters. The methodological validation showed that the linear relationship of the components in rat plasma samples were good( r>0. 995). The recovery rate and matrix effect of plasma samples with low,middle and high concentration were 79. 14%-101. 4%. The intra-day and inter-day precision,accuracy and stability meet the requirements of biological sample analysis. The half-life( t1/2) of chlorogenic acid,gardenioside,oroxylin A did not change significantly and the area under blood concentration-time curve( AUC0-t) is proportional to the dose,which suggested that three components showed a linear kinetic characteristics,but baicalin showed nonlinear kinetic characteristics. Moreover,the retention time of each component in rats was short. The established UPLC-MS/MS quantitative analysis method is rapid,sensitive and accurate,which can be used for the determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma and pharmacokinetic study of Shuganning Injection.

Study on the Isolation and Purification of Baicalin Metabolites from Rat Bile and Its Effect on the Proliferation of Liver Cancer HepG 2 Cells / 中国药房

OBJECTIVE：To isolate and purify baicalin metabolites from rat bile ，and to study its effect on the proliferation of liver cancer HepG 2 cells. METHODS ：Totally of 6 SD rats were collected ，anesthetized and then intubated with bile ducts. They were given baicalin （168 mg/kg）intragastrically after the rats were awake as well as the bile sample 1 was collected during 24 h after intragastric administration. After processed ，bile sample 1 was preliminarily analyzed by HPLC. Another 5 SD rats were anesthetized and intubated in the same way as above ，and then given baicalin intragastrically （400 mg/kg）. The bile sample 2 was collected during 48 h after intragastric administration. After processed ，the bile sample 2 was isolated and purified by semi-preparative HPLC. The isolated metabolites were identified by using UV ，IR，MS and NMR ，as well as based on physical and chemical properties. MTT method combined with high content cell imaging analysis system was used to study the effect of the metabolites on the proliferation of HepG 2 cells. RESULTS ：Baicalin was mainly metabolized into metabolite 1 and metabolite 2 through bile. After isolation and purification ， they were identified as oroxylin A- 7-O-β-D-glucuronide and 2726719792@qq.com baicalein 6-O-β-D-glucuronide. The results of MTT assay showed that the metabolite 2 had a significant inhibitory effect 分析。E-mail：gaoxl@gmc.edu.cn on the proliferation of HepG 2 cells，and its IC 50 was 90 µg/mL；the results of high content cell imaging analysis showed that at a certain concentration metabolite 2 may inhibit proliferation by changing the mitochondrial distribution and membrane permeability of HepG 2 cells（studies on the pharmacological activities of metabolism 1 had been reported ，it was skipped in this study ）. CONCLUSIONS ：In this study ，two baicalin bile metabolites were successfully isolated and identified ，of which baicalein 6-O-β-D-glucuronide has a significant inhibitory effect on the proliferation of HepG 2 cells.

Screening and Identification of Endophytic Fungi in Scutellariae Radix Transformed Baicalin to Baicalein and Oroxylin A / 中国实验方剂学杂志

Objective: In order to explore natural active ingredients of Scutellariae Radix and alleviate the pressure of traditional medicinal resources,this research aimmed to screen endophytic fungus strains from Scutellariae Radix which could transform baicalin into some active ingredients.Method: Taking fresh plants of Scutellariae Radix as strain resource,endophytic fungus strains were isolated by potato dextrose agar (PDA) plate separation,Scutellariae Radix powder selective culture and HPLC detection.Result: An endophytic fungus strain R3 was isolated and it can transform baicalin to baicalein and oroxylin A simultaneously in potato dextrose broth (PDB) contained 0.1% baicalin and the conversion rate reached 61.09% in 5 d cultured at 28℃ and 150 r·min-1,molar ratio of baicalein and oroxylin A was 3:5.With the method of morphological analysis,microscopic identification and 18SrDNA sequence analysis,the bacterium was identified as Penicillium sp.R3. Conclusion: Penicillium sp.R3 can transform baicalin to baicalein and oroxylin A with certain research value and application value.

Oroxylin A ameliorates isoproterenol-induced heart failure model in rats through promoting myocardial autophagy / 中国药科大学学报

@#The protective effect and mechanism of Oroxylin A, a naturally occurring compound in Scutellaria baicalensis Georgi, was investigated in this study. Isoproterenol administration to rats triggered classic cardiac failure, as demonstrated by objective parameters of cardiac dysfunction. Intragastric administration of oroxylin A at the dose of 25, 50 and 100 mg/(kg·d)significantly improved deranged cardiac parameters in the isoproterenol-induced heart failure model in a dose-dependent manner. At the same time, oroxylin A markedly ameliorated cardiac histological changes and down-regulated serum levels of various neuroendocrine factors including norepinephrine, aldosterone, brain natriuretic peptide, endothelin 1, angiotensin II and so on. Mechanistically, augmenting autophagy of myocardial cells via the inhibition of AKT1-RPS6KB1 signaling contributed to the improvement of isoproterenol-induced rat heart failure by oroxylin A. Taken together, these results suggest that oroxylin A ameliorates heart failure through promoting autophagy in myocardial cells.

Inhibitory Effects of Norwogonin, Oroxylin A, and Mosloflavone on Enterovirus 71

Severe complications associated with EV71 infections are a common cause of neonatal death. Lack of effective therapeutic agents for these infections underlines the importance of research for the development of new antiviral compounds. In the present study, the anti-EV71 activity of norwogonin, oroxylin A, and mosloflavone from Scutellaria baicalensis Georgi was evaluated using a cytopathic effect (CPE) reduction method, which demonstrated that all three compounds possessed strong anti-EV71 activity and decreased the formation of visible CPEs. Norwogonin, oroxylin A, and mosloflavone also inhibited virus replication during the initial stage of virus infection, and they inhibited viral VP2 protein expression, thereby inhibiting viral capsid protein synthesis. However, ribavirin has a relatively weaker efficacy compared to the other drugs. Therefore, these findings provide important information that will aid in the utilization of norwogonin, oroxylin A, and mosloflavone for EV71 treatment.

Anti-Allergic Effect of Oroxylin A from Oroxylum indicum Using in vivo and in vitro Experiments

Oroxylum indicum has long been used in Asian traditional medicine to prevent and treat respiratory diseases, diabetes, diarrhea and other conditions. Oroxylin A is a flavone that is present in Oroxylum indicum and in Scutellaria baicalensis. Because the root extracts of both plants have been shown to have anti-allergic effects, the authors investigated whether oroxylin A is likely to have beneficial effects on allergic asthma using female Balb/c mice and rat RBL-2H3 mast cells. Antigen-induced degranulation was measured in vitro by measuring β-hexosaminidase activity. A murine ovalbumin-induced allergic asthma model was used to test the in vivo efficacy of oroxylin A. Sensitization and challenge of ovalbumin induced allergic asthma responses, the accumulations of eosinophils and Th2 cytokine levels in bronchoalveolar lavage fluid and lung tissues. Oroxylin A administration decreased numbers of inflammatory cells, especially eosinophils, and reduced the expression and secretion of Th2 cytokines, including IL-4 and IL-13, in lung tissues and bronchoalveolar lavage fluid. Histologic studies showed oroxylin A reduced inflammatory signs and mucin production in lungs. These findings provide evidence that oroxylin A has potential use as an anti-allergic therapeutic.

Simultaneous determination of baicalin, wogonoside, baicalein, wogonin, and oroxylin-A in Scutellaria baicalensis by UPLC / 中草药

Objective: To establish a UPLC method for the simultaneous determination of five constituents (baicalin, wogonoside, baicalein, wogonin, and oroxylin-A) in Scutellaria baicalensis. Methods: Five constituents in S. baicalensis were simultaneously determined by UPLC with Waters Acquity UPLC BEH C18 column (50 mm × 2.1 mm, 1.7 μm). With a gradient program of acetonitrile-0.1% formic acid at the flow rate of 0.6 mL/min, and the detection wavelength was set at 280 nm. Results: Baicalin, wogonoside, baicalein, wogonin, and oroxylin-A in S. baicalensis were clearly and respectively separated in 15 min. The relationship between the concentration and the peak areas of the five constituents in S. baicalensis was all linear. The RSD of recovery, precision, and reproducibility was all less than 3.0%. Conclusion: The method is rapid, simple, reliable, and accurate, and has been successfully used to the quantification of the five constituents in S. baicalensis.

Effect of oroxylin A on blood-brain barrier permeability induced by hypoxic injury in vitro and its mechanism / 中草药

Objective: To study the protective effect of oroxylin A on blood-brain barrier (BBB) permeability induced by hypoxic injury and its possible mechanism. Methods: The BBB model in vitro was established by primarily culturing brain microvascular endothelial cells (BMEC), and the hypoxic injury was induced. To observe the changes of BBB permeability, fluorospectrophotometry was used to detect the fluorescence intensity of oroxylin A on fluorescein isothiocyanate (FITC)-dextran in BMEC. The effects of oroxylin A on the mRNA expression of ZO-1 and claudin-1 were assayed by RT-PCR, and the effect on the ZO-1 protein expression was measured by Cell-Based ELISA. Results: Hypoxia significantly increased FITC-dextran permeability, decreased the expression of ZO-1 mRNA and protein, while oroxylin A could antagonize these effects caused by hypoxia. Conclusion: Oroxylin A has a protective effect on BBB permeability induced by hypoxia in vitro, and the mechanism is involved in the regulation of ZO-1 expression.

Optimizing Extraction Technology for Total Flavonoid-Aglycone in Scutellaria baicalensis After Enzymatic Hydrolysis by Orthogonal Experiment / 中药新药与临床药理

Objective To study the optimization of extraction technology for total flavonoid-aglycone from Scutellaria baicalensis after enzymatic hydrolysis. Methods With three factors of alcohol concentration, dosage and speed of percolation as the indexes, orthogonal experiment was performed. Concentrations of Baicalein, Wogonin and oroxylin-A were analyzed by HPLC. Results The optimal extraction technology is A2B3C2: extracting the medical material by 10 times(V) 95 % alcohol, the speed being 6 mL/kg per min. Conclusion Under optimal extraction technology, extraction rate of total flavonoid-aglycone is very considerable and the concentration is high to 60 % .