Inhibitory effect of water extract of Sophora flavescens on inflammation and oxidative stress through activating Nrf2/HO-1 signaling pathway / 中草药

Objective: To study the molecular mechanisms of antioxidant effect and anti-inflammatory of water extract of Sophora flavescens (WSF) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Methods: The optimum concentration of WSF was evaluated by CCK-8 assay. The inflammatory model was established with LPS by stimulating RAW264.7 cells in vitro. Then all cells were divided into control group, model group, WSF group and WSF control group. The levels of ROS and NO were analyzed with flow cytometry. Subsequently, the expression of iNOS, COX-2, Nrf2, and HO-1 was detected with qRT-PCR and Western blotting. Finally, the pro-inflammatory cytokines IL-6, TNF-α and anti-inflammatory cytokine IL-10 were detected by ELISA. Results: The CCK-8 assay revealed that 0.01 mg/mL WSF did not affect the cell viability. Compared with control group, the LPS-induced inflammatory response could significantly increase the production of NO and ROS, and the IL-6 and TNF-α were also significantly increased (P < 0.05, 0.01, and 0.001). Furthermore, the expression of iNOS and COX-2 were significantly increased (P < 0.01, 0.001), but the expression of Nrf2 and HO-1 were inhibited (P < 0.05). However, compared with model group, the WSF group not only significantly decreased the levels of NO, ROS, IL-6, and TNF-α, but also decreased the expression of iNOS and COX-2 (P < 0.05, 0.01, and 0.001). In contrast, the the level of IL-10 and the expression of Nrf2 and HO-1 were significantly increased (P < 0.05, 0.01, and 0.001). Conclusion: These results suggested that SF exerted protective effect against LPS-induced inflammatory and oxidative responses in RAW 264.7 cells by the activation of the Nrf2/HO-1 pathway.

Exploration on mechanism of anti-coronavirus of Sophora flavescens through multi-pathway / 中草药

The coronavirus disease 2019 (COVID-19), originated in Wuhan city, Hubei Province, China in December 2019, and then quickly spread to most provinces and regions in China and even spread to many countries abroad. COVID-19 is characterized by wide epidemic, strong infectivity, rapid onset and critical condition. In the face of this epidemic, all parts of the country quickly set off a peak in the fight against COVID-19, but no effective drug for COVID-19 has been developed in the short term. Recently, many hospitals have combined traditional Chinese medicine with western medicine in treatment, and the clinical effect is remarkable, which proves the antiviral effect of traditional Chinese medicine. A large number of pharmacological and clinical studies have proved that the Chinese materia medica S. flavescens has significant antiviral effect. In this paper, the mechanism of anti-coronavirus effect of S. flavescens is expounded from multiple pathways, such as type I interferon, NF-κB signal pathway, ERK signal pathway, PI3K/Akt signal pathway and matrine alkaloids, etc. It is intended to provide reference for clinical treatment of coronavirus infection pneumonia and research and development of related drugs of S. flavescens.

Research progress and prospect on antitumor mechanism of matrine and oxymatrine / 中草药

Sophora flavescens is a traditional herb medicine in China. Matrine and oxymatrine are the primary components in S. flavescens with an outstanding anti-cancer potentials. This review aims to summarize the mechanism of tumor-inhibition of matrine and oxymatrine by screening and analyzing the recent literatures. It was found that the molecular mechanisms of tumor-inhibition of matrine and oxymatrine include the regulation of cell cycle progression, induction of cell apoptosis, anti-metastatic and anti-invasion activities, induction of autophagy of tumor cells, reversion of the multidrug resistance in cancer cells, and regulation of metabolic level in cancer cells. Further research on the molecular mechanisms of matrine and oxymatrine could reveal their cellular signaling network and gene regulation mechanism in the anticancer behaviors, so as to find more accurate tumor therapeutic targets, which have important implications not only for new drugs development in clinical application of cancers, but also for the modernizations and internationalization of Chinese medicine.

UPLC-MS/MS-based pharmacokinetics of self-microemulsion containing total flavonoids from Sophora flavescens in rats / 中草药

Objective: To establish a UPLC-MS/MS method for simultaneous determination of sophoraflavanone G (SFG), kurarinone (Kur), and isoxanthohumol (Iso) in rat plasma using rutin as the internal standard (IS), and to study the pharmacokinetic parameters of total flavonoids (TF) from Sophora flavescens and TF self-microemulsion drug delivery system (TF-SMEDDS) in rats. Methods: Analysis was carried out on an ACQUITY UPLC® HSS T3 C18 column using acetonitrile-0.1% formic acid in water as the mobile phase at a flow rate of 0.2 mL/min. The plasma samples were prepared by liquid-liquid extraction with ethyl acetate. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring with an electro-spray ionization source in negative ionization mode. All statistical analysis was performed using DAS 2.0 software package. Results: The calibration curves of Kur, SFG, and Iso exhibited good linearity (r2 > 0.995 9) over the range of (0.02-1 600.00), (0.015-1 200.000), and (0.01-800.00) ng/mL, respectively. Precision, accuracy, average extraction recovery, and matrix effects were all in line with biological sample analysis requirements. The pharmacokinetic parameters of Kur, SFG, and Iso from TF were as follows: t1/2z (7.04 ±2.46), (4.54 ± 2.13), (4.73 ± 1.67) h, and AUC0-∞ (3 469.57 ± 555.37), (2 524.48 ± 425.83), (1 006.47 ± 85.46) ng∙h/mL. The pharmacokinetic parameters of Kur, SFG, and Iso from TF-SMEDDS were as follows: t1/2z (13.10 ± 2.67), (11.47 ± 4.17), and (12.67 ± 4.97) h, and AUC0-∞ (13 002.49 ± 2 498.09), (8 070.80 ± 2 264.62), (3 918.85 ± 429.76) ng∙h/mL. The relative bioavailabilities of Kur, SFG, and Iso in TF-SMEDDS were approximately 374.76%, 319.70%, and 389.37%, respectively. Conclusion: The UPLC-MS/MS method can be used to study the pharmacokinetics of three components in S. flavescens in rats. The bioavailability of TF-SMEDDS in rats was significantly increased.

Molecular mechanism of Danggui Beimu Kushen Pills in treating prostatic diseases based on TCM-IP / 中草药

Objective To analyze the material basis and molecular mechanisms of Danggui Beimu Kushen (DBK) Pills in treating prostatic diseases based on the method of integrated pharmacology. Method The platform of Integrative Pharmacology of Traditional Chinese Medicine (TCM-IP, www.tcmip.cn) was utilized to predict the main active ingredients and functional targets of DBK Pills in treating prostatic disease, key targets were screened for enrichment analysis of pathways, and the network of “herb-core component-key target-main pathway” was constructed, and the possible mechanisms of DBK Pills in treating prostatic diseases were explored. Results A total of 532 candidate key targets for the treatment of prostatic diseases by DBK Pills were predicted, and 1 840 terms of gene function and 194 signal pathways were analyzed by gene ontology (GO) and KEGG, respectively. The network analysis of “herb-core component-key target-main pathway” showed that 65 core components were predicted, including 29 ingredients from Angelica sinensis, 11 from Fritillaria thunbergii and 26 from Sophora flavescens. Those predicted components acted on the key targets of prostatic diseases, such as transcription factor binding, negative regulation of apoptosis, et al, through the estrogen, apoptosis, chemokines and other signal pathways, and thus played a role in the regulation of cell cycle, apoptosis and proliferation imbalance, which might be the molecular mechanisms of DBK Pills for the treatment of prostatic disease. Conclusion DBK Pills regulate the development of BPH, prostate cancer and other diseases through multiple pathways with multi-component interacting with multiple targets.

Simultaneous Determination of Seven Alkaloid Constitutents in Sophora Flavescens Ait. by HPLC-DAD / 中国药师

Objective:To develop an HPLC-DAD method for the simultaneous determination of seven alkaloid constitutents, so-phoranholN-oxide, oxymatrine, sophoridine, oxysophocarpine, sophoranol, matrine and sophocarpine in Sophora flavescens Ait. Meth-ods:The chromatographic separation was performed on an Agilent Zorbax C18 column (150 mm × 4. 6 mm, 5μm) with 10 mmol·L-1 NH4Ac(0.1% ammonia, pH=9.0)(A)-acetonitrile-methanol(1 ∶22) containing 20 mmol·L-1NH4Ac(B) as the mobile phase with gradient elution at the flow rate of 1 ml·min-1 . The detection wavelength was set at 220 nm,and the column temperature was maintained at 30 ℃. Results: SophoranholN-oxide, oxymatrine, sophoridine, oxysoph-ocarpine, sophoranol, matrine and sopho-carpine was linear within the range of 0.093-1.860 μg(r =0.999 6) , 0.530-10.600 μg (r =0.999 7), 0.062-1.240 μg (r =0. 999 8), 0. 281-5. 620 μg (r=0. 999 9), 0. 026-0. 520 μg (r=0. 999 8) ,0. 036-0. 720 μg (r=0. 999 7) and 0. 032-0. 640 μg (r=0. 999 6), respectively. The average recovery was 97. 5%, 98. 2%, 99. 0%, 99. 4%, 99. 2%, 98. 2% and 98. 7%, and RSD was 1. 18%, 0. 92%, 1. 43%, 1. 04%, 0. 81%, 0. 43% and 0. 88%(n =6), respectively. Conclusion:The method is convenient, accurate and reproducible in the quality control of multicomponents in Sophora flavescens Ait.

Application of electronic tongue in pattern study about bitterness inhibition by hydroxypropyl-β-cyclodextrin / 中草药

Objective To study the bitterness inhibition law of hydroxypropyl-β-cyclodextrin (HP-β-CD) concentration (C) on the bitter compounds and bitter Chinese herbal medicine, and to explore the application of electronic tongue in this study. Methods Berberine, oxymatrine, Sophora flavescens, and Andrographis paniculata decoction were used as bitterness vectors to establish two models of bitterness inhibition law about ΔI-C and ΔIe-C, and to explore the prediction model of bitterness inhibition effect about ΔI-ΔIe, based on the oral taste evaluation results (ΔI) and electronic tongue information (ΔIe). Then, fitting precision and fitting goodness of the prediction model were evaluated with cross-validation and residual analysis. Results In this study, good Weibull model of bitterness inhibition pattern about ΔI-C were established for all the four bitterness vectors, the decision coefficient R2 are as followed: 0.999 6, 0.987 9, 0.996 4, and 0.998 4 (P < 0.01); The decision coefficient R2 of six (two sensors per vector) models of bitterness inhibition law about ΔIe-C of berberine, S. flavescens, and A. paniculata decoctions were as followed: 0.996 5, 0.991 6, 0.997 3, 0.989 3, 0.999 6, and 0.999 1 (P < 0.01); The decision coefficient R2 of six corresponding linear prediction models of bitterness inhibition effect about ΔI-ΔIe were as followed: 0.989 1, 0.968 3, 0.989 0, 0.982 0, 0.977 9, and 0.986 1 (P < 0.01); The correlation coefficient R calculated by correlation coefficient of six prediction models above were as followed: 0.986 0, 0.997 3, 0.988 4, 0.960 8, 0.980 2, and 0.983 9 (P < 0.01); No model was established for oxymatrine within the range of tested concentration in this research, as it didn’t respond to the four sensors with varied concentration. Conclusion Based on this method, the bitterness inhibition law of HP-β-CD with changed concentration was obtained. Prediction models based on HP-β-CD concentration or electronic tongue data were also established, they can be used to predict the relative bitterness inhibition effect. Part of the bitter compounds didn't response to the electronic tongue regularly, remain further research and development of electronic tongue technology.

Optimization of nanofiltration concentration process of Sophora flavescens extract by coupling technology of response surface methodology and regulating Donnan effect / 中草药

Objective: Response surface analysis and regulating Donnan effect methodology were used to optimize the matrine concentration by nanofiltration techonology. Methods: On the basis of single factor experiment results, molecular weight cut-off (MWCO) of nanofiltration membrane, concentration and pH were selected as influencing factors to evaluate the retention rate of matrine and total alkaloids with Box-Behnken central composite experiment design, and then, the optimal concentration parameters were calculated in the conditions of pH 6-7 to regulate Donnan effect between alkaloids and nanofiltration membrane. Results: The retention rate of matrine was of positive relevance with the ethanol concentration. The optimal concentration parameters were as follows: cutting off molecular weight of 150, pH of 6.19, concentration of 204.3 μg/mL, ethanol concentration of 15%, the retention rate of matrine and total alkaloids were 94.41% and 97.63%, respectively. Conclusion: The combination of ethanol regulation Donnan effect and response surface analysis can well optimize the concentration process of S. flavescens extract by nanofiltration, and the results provide the references for nanofiltration concentration for heat-sensitive Chinese materia medicia.

Simultaneous determination of berberine, matrine and oxymatrine in traditional Chinese medicines by using nonaqueous capillary electrophoresis / 药物分析学报

A rapid method for the simultaneous determination of berberine (BBR), matrine (MT) and oxymatrine (OMT) by nonaqueous capillary electrophoresis (NACE) was developed. Optimum separation of the analytes was obtained on a 50cm×50μm i.d. fused-silica capillary using a non-aqueous buffer system of 70mM ammonium acetate, 7.0% acetic acid and 10% acetonitrile at 25kV and 20℃. The relative standard deviations (R.S.D.) of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berberine, 0.11%-0.60% and 0.74%-1.63% for matrine, 0.15% and 0.45% for oxymatrine, respectively. Detection limits of berberine, matrine and oxymtrine were 0.18μg/mL, 4.08μg/mL and 4.16μg/mL, respectively. In the tested concentration range, good linear relationships (0.9992 for berberine, 0.9988 for matrine and 0.9988 for oxymatrine) were observed. The linear calibration ranges were 0.45-360.0μg/mL for berberine, 8.16-408.0μg/mL for matrine and 20.8-416.0μg/mL for oxymatrine. This method has been successfully applied to the phytochemical analysis of alkaloids extracts from two commonly used traditional Chinese herbal drugs: Sophora flavescens Ait. (Kushen) and Cortex phellodendri chinensis (Huangbai) and their medicinal preparations.

Simultaneous determination of berberine, matrine and oxymatrine in traditional Chinese medicines by using nonaqueous capillary electrophoresis / 西安交通大学学报·英文版

A rapid method for the simultaneous determination of berberine (BBR), matrine (MT) and oxymatrine (OMT) by nonaqueous capillary electrophoresis (NACE) was developed. Optimum separation of the analytes was obtained on a 50 cm x 50 μm i. d. fused-silica capillary using a non-aqueous buffer system of 70 mM ammonium acetate, 7.0% acetic acid and 10% acetonitrile at 25 kV and 20°C . The relative standard deviations (R. S. D.) of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berberine, 0.11%-0.60% and 0.74%-1.63% for matrine, 0.15% and 0.45% for oxymatrine, respectively. Detection limits of berberine, matrine and oxymtrine were 0.18 μg/mL, 4.08 μg/mL and 4.16 μg/mL, respectively. In the tested concentration range, good linear relationships (0.999 2 for berberine, 0.998 8 for matrine and 0.998 8 for oxymatrine) were observed. The linear calibration ranges were 0.45-360.0 μg/mL for berberine, 8.16-408.0 μg/mL for matrine and 20.8-416.0 μg/mL for oxymatrine. This method has been successfully applied to the phytochemical analysis of alkaloids extracts from two commonly used traditional Chinese herbal drugs: Sophora flavescens Ait. (Kushen) and Cortex phellodendri chinensis (Huangbai) and their medicinal preparations.

Study of Quality Standard for Qingfei Yihuo Tablet / 中国中医药信息杂志

Objective To establilsh the quality standard for Qingfei Yihuo tablet. Methods TLC was performed to identify Sophora flavescens Ait, Peucedani and Cortex Phellodendri. HPLC was used to determine Baicalin. ODS column was used. The mobile phase consisted of methanol-water-phosphoric acid (47∶53∶0.2). The flow rate was 1.0 mL/min and column temperature was at 25 ℃. The detecting wave length was at 280 nm. Results The characteristic for identification by TLC was distinct and hightly specific. Baicalin could be determined by HPLC. The linearity of Baicalin was good in the range of 0.163～0.975 ?g (r=0.999 9). The average recovery of Baicalin was 97.43% and RSD was 1.97% (n=6). Conclusion The methods of identification and quantification is simple and reproducible. It can be used effectively for the quality control of Qingfei Yihuo table.

Adsorption properties of AB08 resin for purification of total flavone in Sophora flavescens / 中草药

Object To study the adsorption function of macroporous resin AB 8 for the purification of total flavones in Sophora flavescens Ait and the effects of concentration, pH, flow rate of the adsorbate and eluant Methods UV spectrophotometry was used to analyze the content of total flavone in S flavescens Results The appropriate adsorption conditions were: concentration of Sophora extract, 0 285 mg/mL ; pH=4; flow rate 3 BV/h, when 50% alcohol was used as eluant, the effect of desorption was satisfactory Conclusion AB 8 resin can be used to refine the total flavone in S flavescens

Influence of producing area and plant age on oxymatrine content in root of Sophora flavescens / 中草药

Object The influence of producing area and plant age on oxymatrine levels in the dried root of Sophora flavescens Ait. has been investigated to provide useful information to optimize the areas for mass scale propagation, proper management and species conservation. Methods The oxymatrine was quantified by HPLC and various statistical treatments were carried out following SPSS 9.0 and Microsoft Excel. Results The oxymatrine concentrations in the root samples collected from 17 provinces and regions differed from 0.494% to 4.127% and the maximum oxymatrine content (3.493%) was recorded in the root sample collected from Heilongjiang Province. Analysis of variance indicates a highly significant difference in the oxymatrine content of roots collected from different provinces, and the samples from the cold arid northern high-latitude areas had higher oxymatrine than those from warm humid southern low-latitude areas in the mainland China. Plant age is positively related to the total root biomass and oxymatrine content. Conclusion The oxymatrine content in the roots of S. flavescens is quite different between different areas and plant ages. The S. flavescens growing in the cold arid northern high-latitude areas had higher oxymatrine concentration.

Study on extraction of Sophora flavescens Ait. by HPLC-MS / 中成药

Objective:Chemical constituents of Sophora flavescems Ait. were studied by different extraction. Methods: Online HPLC/ESI/MS was used to study the different extraction. Results: numbers of alkaloids and flavone from Sophora flavescens Ait. were 9,8 and 12, respectively by means of chloroform strong aqua , water , and water methanol extraction. Conclusions: For the first time, we studied extraction of Sophora flavescems Ait by HPLC MS. This provided basic for study on fingerprints of Sophora flavescems Ait..