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A high performance liquid chromatography charged aerosol detector (HPLC-CAD) method was established for the simultaneous determination of five saponins (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, Ginsenoside Rd) in Yaobitong capsule, providing a method for quality control. The sample was extracted with methanol and chromatographic separation was performed on a Waters Xbridge Phenol column (150 mm×4.6 mm, 3.5 μm) using acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1.0 mL·min-1. The column temperature was 30 ℃ and the injection volume was 10 μL. The nebulizer temperature of CAD was 35 ℃ and the air pressure was 60.2 psi, the filtration was 3.6 s, and the collection frequency was 5 Hz. Notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd showed a good linear relationship in the range of 16.96-203.5 μg·mL-1 (R2 = 0.999 3), 54.46-653.5 μg·mL-1 (R2 = 0.999 3), 10.96-131.5 μg·mL-1 (R2 = 0.999 6), 51.50-618.0 μg·mL-1 (R2 = 0.999 0), 15.94-191.3 μg·mL-1 (R2 = 0.999 4), respectively. The average recoveries were 98.96%, 100.8%, 94.76%, 100.1%, 103.1%, and RSDs were 0.87%, 1.46%, 1.85%, 2.06%, 0.96% (n = 6), respectively. The proposed method is accurate, simple and reliable, and can be used for the determination of five saponins in Yaobitong capsule.
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Objective To investigate the chemical constituents from the ethanol extract of Yaobitong Capsule. Methods The chemical constituents were isolated by repeated silica gel column, dynamic axial compression chromatography, Sephadex LH-20 column, and prep-HPLC. Their structures were identified on the basis of NMR spectral data analysis. Results Sixteen compounds were isolated and identified to be nodakenetin (1), columbianetin (2), ferulic acid (3), umbelliferone (4), bergaptol (5), xanthotoxin (6), isoimperatorin (7), scoparone (8), (Z)-6-hydroxy-7-methoxydihydroligustilide (9), 4-hydroxy-3-butylphthalide (10), 3-(3′-hydroxy)- butylphthalide (11), senkyunolide C (12), senkyunolide H (13), senkyunolide I (14), fraxidin (15), and isofraxidin (16). Conclusion Compounds 1—16 are isolated from Yaobitong Capsule for the first time, and compound 11 is obtained by the means of separation for the first time.
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Objective To investigate the anti-inflammatory constituents from the ethanol extract of Yaobitong Capsule. Methods The chemical constituents were isolated by repeated silica gel column, Sephadex LH-20, and preparative HPLC. The structures were identified by various spectral data analysis and physicochemical properties. The anti-inflammatory effects were evaluated by in vitro model of LPS-stimulated rat thoracic aortic smooth muscle cells (A7r5). Results Eleven compounds were isolated from the contents of the capsule, including 2 organic acids, 1 alkaloid, 2 senkyunolides, 1 sesquiterpene, and 5 coumarins, which identified as ethyl caffeate (1), ferulic acid (2), corydaline (3), senkyunolide H (4), senkyunolide I (5), ligustiphenol (6), columbianetin (7), ulopterol (8), meranzin hydrate (9), angelitriol (10), and 6-[1(R),2(R)-1,2,3-thihydroxy-3-methylbutyl]-7-methoxycoumarin (11). Compounds 1-6 showed the anti-inflammatory activity in different degrees. Conclusion Compounds 1, 3, 6, and 8-11 are isolated from Yaobitong Capsule for the first time, and organic acids and senkyunolide compounds may be the anti-inflammatory pharmacodynamic material bases of Yaobitong Capsule.
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Objective The main metabolites of Yaobitong Capsule (YC) in rat urine and bile were investigated by UPLC-Q-TOF- MS/MS, and their major metabolic pathways were summarized. Methods To compare with the control samples, the possible metabolites were found. Their structures were speculated by exact mass and fragment ions. Results YC had 31 metabolites in urine and 35 metabolites in bile under positive and negative ion mode. They were mainly the metabolites of ligustilide, 3-butyl-phthalide, osthole, senkyunolide A, sankyunolide A, emodin, decursinol/marmesin/columbianetin, corypalmine/ isocorypalmine, corybulbine/isocorybulbine/coryphenanthrine, allocryptopine, ferulic acid, cryptochlorogenic acid, angelol and so on. Their major metabolic pathways included hydrolysis of the lactone ring, hydroxylation, demethylation, dehydration, dehydrogenation, dehydroxylation, hydrogenation, methylation, methyl oxidation, glucuronidation, sulphate-binding, N-acetylcysteine-binding, cysteine-binding, cysteinyl acid-glycine-binding, glutathione-binding and so on. In addition, three prototype components, namely paeoniflorin, albiflorin, and ginsenosides Rg1, were detected in rats’ bile. Conclusion The established UPLC-Q-TOF-MS/MS method can analyze the metabolites of YC capsule in rat urine and bile. The in vivo metabolic characteristics of YC have been initially clarified. The results provide a foundation for elucidating the in vivo efficacy ingredients of YC.
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OBJECTIVE@#To discuss the effect of Yaobitong capsule on histomorphology of dorsal root ganglion and on expression of p38MARK phosphorylation in autologous nucleus pulposus transplantation model of rats.@*METHODS@#A total of 60 SD rats were randomly divided into the blank group, model group and Yaobitong capsule group, with 20 rats in each group. The animal model of autologous nucleus pulposus transplantation around the lumbar nerve root was built. Three days after the modeling, rats were given the drugs for the first time, while rats in the model group were given the equivalent normal saline. After 30 d of continuous administration, samples were collected from rats. HE staining was performed on the dorsal root ganglion of L4 and L5 spinal cord of rats in each group and the expression of p38MARK phosphorylation was measured. All data were treated with the statistical analysis.@*RESULTS@#The histological examination showed that the histomorphology of dorsal root ganglion in the Yaobitong capsule group was more significantly improved than the one in the model group, while the results of western blot showed that Yaobitong capsule could significantly inhibit the level of p38MAPK phosphorylation of dorsal root ganglion cells.@*CONCLUSIONS@#Yaobitong capsule can improve the symptoms and nerve radiculopathy of autologous nucleus pulposus transplantation of rats and its mechanism may be associated with its inhibiting effect on the level of p38MAPK phosphorylation.
ABSTRACT
Objective: To discuss the effect of Yaobitong capsule on histomorphology of dorsal root ganglion and on expression of p38MARK phosphorylation in autologous nucleus pulposus transplantation model of rats. Methods: A total of 60 SD rats were randomly divided into the blank group, model group and Yaobitong capsule group, with 20 rats in each group. The animal model of autologous nucleus pulposus transplantation around the lumbar nerve root was built. Three days after the modeling, rats were given the drugs for the first time, while rats in the model group were given the equivalent normal saline. After 30 d of continuous administration, samples were collected from rats. HE staining was performed on the dorsal root ganglion of L4 and L5 spinal cord of rats in each group and the expression of p38MARK phosphorylation was measured. All data were treated with the statistical analysis. Results: The histological examination showed that the histomorphology of dorsal root ganglion in the Yaobitong capsule group was more significantly improved than the one in the model group, while the results of western blot showed that Yaobitong capsule could significantly inhibit the level of p38MAPK phosphorylation of dorsal root ganglion cells. Conclusions: Yaobitong capsule can improve the symptoms and nerve radiculopathy of autologous nucleus pulposus transplantation of rats and its mechanism may be associated with its inhibiting effect on the level of p38MAPK phosphorylation.