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Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after long-term use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an herb pair.However,whether RPA can alleviate RB-induced hepatotoxicity remain unclear.In this work,the results confirmed that RB had a dose-dependent antidepressant effect,but the optimal antidepressant dose caused hepatotoxicity.Notably,RPA effectively reversed RB-induced hepatotoxicity.Afterward,the mechanism of RB-induced hepatotoxicity was confirmed.The results showed that saiko-saponin A and saikosaponin D could inhibit GSH synthase(GSS)activity in the liver,and further cause liver injury through oxidative stress and nuclear factor kappa B(NF-KB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Furthermore,the mechanisms by which RPA attenuates RB-induced hepatotoxicity were investigated.The results demonstrated that RPA increased the abundance of intestinal bacteria with glycosidase activity,thereby promoting the conversion of saikosaponins to sai-kogenins in vivo.Different from saikosaponin A and saikosaponin D,which are directly combined with GSS as an inhibitor,their deglycosylation conversion products saikogenin F and saikogenin G exhibited no GSS binding activity.Based on this,RPA can alleviate the inhibitory effect of saikosaponins on GSS activity to reshape the liver redox balance and further reverse the RB-induced liver inflammatory response by the NF-κB/NLRP3 pathway.In conclusion,the present study suggests that promoting the conversion of saikosa-ponins by modulating gut microbiota to attenuate the inhibition of GSS is the potential mechanism by which RPA prevents RB-induced hepatotoxicity.
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This study analyzed the effect of the Radix Bupleuri-Radix Paeoniae Alba herb pair on endogenous metabolites in rats with chronic unpredictable mild stress (CUMS)-induced depression by using LC-MS liver metabolomics. Rats were randomly divided into 5 groups: a normal control group, a CUMS model group, a venlafaxine-positive group, and a high-low dose group for the Radix Bupleuri-Radix Paeoniae Alba herb pair, with continuous modeling and administration over 28 days. The efficacy of Radix Bupleuri-Radix Paeoniae Alba herb pair was evaluated by measuring traditional pharmacodynamic indicators of depression (body weight, open field test, sucrose preference test and forced swimming test). Animal experimentation was approved by the Committee on the Ethics of Animal Experiments of Shanxi University (SXULL2016036). Liver metabolic profiles were obtained by the UHPLC-Q Exactive Orbitrap-MS metabolomics technique. The results show that the Radix Bupleuri-Radix Paeoniae Alba herb pair can significantly decrease depression-like behavior of rats in the CUMS model group. Increases in 25 depression-related metabolites were identified by LC-MS metabonomics, and the Radix Bupleuri-Radix Paeoniae Alba herb pair could significantly decrease 16 of them. Metabolic pathway analysis showed that D-glutamine and D-glutamate metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, and glutathione metabolism were the main metabolic pathways altered by this herb pair in CUMS model rats.
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We investigated the potential hepatoprotective effect of Radix Bupleuri (RB) by inducing acute liver injury (ALI) in an animal model using acetaminophen (APAP) after pretreatment with RB aqueous extract for three consecutive days. Compared to those of the APAP group, the biochemical and histological results of the RB pretreatment group showed lower serumaspartate transaminase (AST) and alanine transaminase (ALT) levels as well as less liver damage. Pharmacokinetic study of the toxicity related marker acetaminophen-cysteine (APC) revealed a lower exposure level in rats, suggesting that RB alleviated APAP-induced liver damage by preventing glutathione (GSH) depletion. The results of cocktail approach showed significant inhibition of CYP2E1 and CYP3A activity. Further investigation revealed the increasing of CYP2E1 and CYP3A protein was significantly inhibited in pretreatment group, while no obvious effect on gene expression was found. Therefore, this study clearly demonstrates that RB exhibited significant protective action against APAP-induced acute live injury via pretreatment, and which is partly through inhibiting the increase of activity and translation of cytochrome P450 enzymes, rather than gene transcription.
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OBJECTIVE:To establish the quality standard of Chailing hugan granules. METHODS:TLC was used for qualitative identification of Radix bupleuri,Atractylodes macrocephala and Glycyrrhiza uralensis. The content of total flavonoids (by rutin) in preparation was determined by UV-visible spectrophotometry. The moisture, dissolubility and granularity of preparation were determined. RESULTS:TLC spots of R. bupleuri ,A. macrocephala and G. uralensis were clear and well-separated without interference from negative control. The linear range of rutin was 0.050-0.300 mg/mL(r＝0.999 8). RSDs of precision, stability and reproducibility tests were all lower than 2%. The recoveries were 97.89%-100.01%(RSD＝0.68%,n＝9). The content of samples were 1.920-2.018 mg/g. The contents of moisture in 3 batches of samples were 1.54%,1.62% and 1.57%;all samples dissolved within 5 min. The sum of granules not passing through No.1 sieve and passing through No.5 sieve were 2.13%, 2.51%,2.38%,which were all in line with the requirements of Chinese Pharmacopeia (2015 edition,Vol Ⅳ). CONCLUSIONS:The content of total flavonoicle in Chailing hugan granules should be no less than 1.57 mg/g (by rutin). Established quality standard is simple,rapid,accurate and reproducible,and can be used for quality control of Chailing hugan granules.
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Objective: To evaluate the quality of Radix bupleuri from different producing areas and make a comparison by using grey correlation analysis(GRA)and principal components analysis(PCA).Methods: Fifteen samples of Radix bupleuri were collected from different producing areas.The percentage of total ash, acid-insoluble ash, extract, saikosaponin a and saikosaponin d of Radix bupleuri from different habitats were determined.The grey correlation quality evaluation model was built for Radix bupleuri from different producing areas and the results were compared with those of PCA.Results: According to the grey correlation analysis on components in the different batches of Radix bupleuri , the samples from Shaanxi province were the most highly correlated with the optimal reference sequence (0.79), which suggested the best quality.Radix bupleuri Fructus from Hebei province was the most poorly correlated with the optimal reference sequence (0.23), which showed the worst quality.The results were basically identical with those of PCA, indicating good repeatability and stability.Conclusion: Grey incidence degree method and the established Radix bupleuri quality evaluation models can be used to evaluate the quality of Radix bupleuri .
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OBJECTIVE: To explore the molecular mechanism and targets with"Relieving the Depressed Liver" effectofemotional diseases treatment by Radix Bupleuri. METHOD: Systems pharmacology methodwas employed to exploring the ingredients, targets and diseases information which was related "Relieving the Depressed Liver" effect in Radix Bupleuri. Next, the "disease-target" network and "drug-target" network were built by Cytoscape 3.2.1 and we analyzed the network by using network topological analysis method and systems pharmacological data. RESULT: Twenty-five targets were found to be associated with emotional diseases. Eight core ingredients from 118 ingredients in Radix Bupleuri were directly related to these targets. The molecular mechanism of Radix Bupleuri treatment of emotional diseases is produced by the interactions between the 118 ingredients and 25 potential targets, and eight core compounds may play a central role in the process of this treatment mechanism. CONCLUSION: Through the construction of systems pharmacology models, the molecular mechanism of "Relieving the Depressed Liver" effect of Radix Bupleuri from the system level is we interpreted. Our study provides a theoretical foundation for the explanation of TCM treating mechanism and "treat different diseases with same Chinese medicine" phenomenon.
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OBJECTIVE: To establish a method for determining the contents of saikosaponin a, b1, b2 and c in Radix Bupleuri formula granules by HPLC-MS/MS, and provide scientific basis for the quality standards. METHODS: The samples were separated on a Kinetex C18 column (2.1 mm×100 mm, 2.6 μm) by gradient elution at the flow rate of 300 μL·min-1 using acetonitrile and 0.1% aqueous formic acid as the mobile phase. The column temperature was maitained at 40℃. Multiple reaction monitoring scanning (MRM) was employed for the quantification with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at -4 500 V and the turbo spray temperature was maintained at 550℃. RESULTS: There was significant correlation between the peak area and concentration of each compound within the test range (r≥0.999 1). The average recoveries were from 97.44% to 100.56%. CONCLUSION: Saikosaponin a, b1, b2 and c are determined by this method simultaneously. The method is available with good reproducibility and can control the quality of Radix Bupleuri formula granules effectively.
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OBJECTIVE:To study the spectrum-effect relationship of the antipyretic effect of Radix Bupleuri injection. METH-ODS:HPLC method was used to establish the fingerprint of Radix Bupleuri injection. The antipyretic effect of 10 batches of Radix Bupleuri injection on fever rats induced by dried yeast were determined respectively. The fingerprint peaks were screened,along with the temperature value of different time points(0,10,15,30,40,55,70 min). The principal components were extracted by principal component method and the indirect relationship between the fingerprint and antipyretic effect was analyzed,depending on the principal component correlation coefficient matrix. RESULTS:There were 39 common peaks in fingerprint(similarity>0.85), No. 8,12,14,19,26,31,34,35 and 39 common peaks with large peak area were included in the study. Four principal compo-nents were extracted by principal component analysis(87% of total variant). First principal component showed that the active com-ponents of the 12th peak may be related to the antipyretic effect of 6 to 13 hours. The second principal component showed that the active components of the 26th peak may be related to the antipyretic effect of 0.5 to 5 hours. The third principal component showed that the similar effect of the active components could be caused by 34th,35th and 39th peaks. The fourth principal component sug-gested that there were some similarities between the 14th and the 31st peaks. CONCLUSIONS:Radix Bupleuri injection have obvi-ous improvement for fever rats. There is certain corresponding relation between HPLC fingerprint and antipyretic effect of Radix Bu-pleuri injection .
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OBJECTIVE: To establish the determination method of acyl-saikosaponin in Radix Bupleuri. METHODS: Single factor analysis and orthogonal experimental design were used to optimize the acyl-saikosaponin hydrolysis parameters such as concentration of aqueous ammonia, time of ultrasound-assisted hydrolysis and ultrasound power by L9 (34) orthogonal test. An HPLC method was employed to determine the saikosaponins contents, which was carried out on a WondaSil C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase of methanol-water (53:47) at the flow rate of 1.0 mL · min-1 and the wavelength of 210 nm. RESULTS: The optimized conditions of acyl-saikosaponin hydrolysis were as follows: the concentration of aqueous ammonia was 23%, the time of ultrasound-assisted hydrolysis was 25 min, and the ultrasound power was 350 W. The content of acyl-saikosaponin in the Radix Bupleuri samples purchased in 12 different drug stores was lower than that of saikosaponin in Radix Bupleuri. CONCLUSION: This method is simple and reliable, which can be used for the determination of acyl-saikosaponin in Radix Bupleuri and provides clues for the quality assessment of Radix Bupleuri.
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This study was aimed to investigate the effect of Radix Bupleuri extract (RBE) on 5-HT3R channel currents of primarily cultured hippocampal neurons in depression emotion rats.Depression emotion model ratswere duplicated.RBE was used for drug intervention.And then,the rats were evaluated by the open-field test (OFT) and the sucrose preference test.Serum of rats in each group was collected and then added into the primary cultured hippocampal neurons for 24 h.The 5-HT3R channel currents were recorded by the whole-cell patch clamp.The results showed that compared with the normal group,the total score of OFT in the model group was significantly decreased (P < 0.01); the sucrose preference ratio decreased obviously (P < 0.01); and the current density value of primary cultured hippocampal neurons in serum of the model group was significantly higher (P <0.01).Compared with the model group,the total scores of OFT in the RBE group and fluoxetine group increased significantly (P < 0.05,P < 0.01); the sucrose preference ratio also increased obviously (P < 0.05,P < 0.05); and current density value of the primary cultured hippocampal neurons in serum of the RBE group and fluoxetine group decreased significantly (P < 0.01,P < 0.01).It was concluded that RBE can effectively correct the abnormal 5-HT3R channel currents of rats with depression emotion,which may be one of the central mechanisms in the treatment of depression emotion.
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OBJECTIVE: To establish a rapid and nondestructive method to determine coating thickness of the total saponin of Radix Bupleuri enteric coated tablets by near infrared spectroscopy (NIRS) analytical technique. METHODS: Spectral pretreating methods, wavenumber selection and factors of NIRS were discussed in detail. Different modeling methods were compared and the methodology of model was investigated. RESULTS: Partial least squares regression (PLSR) was used for building the quantitative calibration model. Correlation coefficients(r), root-mean-squares error of cross-validation (RMSECV), and root mean square error of prediction (RMSEP) obtained by PLSR model were 0.9915, 3.6 and 3.24 μm respectively. CONCLUSION: Predicted results show that the established method is rapid, nondestructive, and reliable, which can be applied to the online monitor of Chinese medicine tablets coating process.
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OBJECTIVE: To study the dissolving behaviors of saikosaponin a and d, a pair of epimeride and the main effective substances in Radix Bupleuri, during the decoction.
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OBJECTIVE: To establish an HPLC method for determination of five saikosaponins in the leftovers from the preparation process of Radix Bupleuri Oral Liquid. METHODS: The five constituents were analyzed on an Hypersial C18 column (4.6 mm × 250 mm, 5 μm) by gradient elution using acetonitrile-water as the mobile phase. The flow rate was 1.0 mL · min-1. The column temperature was 30°C. The detection wavelengths were set at 210 and 254 nm. RESULTS: The calibration curves were linear in the ranges of 0.23-4.6 μg (r=0.9998), 0.11-2.2 μg (r=0.9956), 0.15-3.0 μg (r=0.9996), 0.35-3.5, μg (r=0.9993) and 0.71-7.1 μg (r=0.9997) for saikosaponin a,c,h,b1 and b2, respectively. The average recoveries of saikosaponin a, c, h, b1 and b2 were 98.10% with RSD of 1.73%, 100. 60% with RSD of 2.25%, 98.60% with RSD of 2.08%, 97.27% with RSD of 0.83% and 99.70% with RSD of 2.26%, respectively. CONCLUSION: This method is simple and reliable, which can be used for the determination of saikosaponin a, c, h, b1 and b2 in the leftovers from the preparation process of Radix Bupleuri Oral Liquid. Copyright 2012 by the Chinese Pharmaceutical Association.
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An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector （HPLC-DAD）. Similarity and hierarchical clustering analysis （HCA） were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory, and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis, which was one of the main ingredients of Xiaochaihu granules, was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time （RT） and UV spectrum feature similarity of their corresponding peaks. Eleven matching peaks were found between Xiaochaihu granules and Radix Bupleuri Chinensis. However, the saikosaponin A and saikosaponin D, which are the important active components in Radix Bupleuri Chinensis, were not found in Xiaochaihu granules from any manufacturers. The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15. The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category. Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category. The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components. These active components, existing in raw herb, might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology. So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae, but also provide some guidance for preparation technology of Xiaochaihu granules.
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An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector (HPLC-DAD).Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory,and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis,which was one of the main ingredients of Xiaochaihu granules,was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks.Eleven matching peaks were found between Xiaochalhu granules and Radix Bupleuri Chinensis.However,the saikosaponin A and saikosaponin D,which are the important active components in Radix Bupleuri Chinensis,were not found in Xiaochaihu granules from any manufacturers.The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15.The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category.Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category.The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components.These active components,existing in raw herb,might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology.So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae,but also provide some guidance for preparation technology of Xiaochaihu granules.
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OBJECTIVE:To determine the content of volatile oil and saikosaponins in Radix Bupleuri collected in different harvesting seasons.METHODS:Volatile oil was extracted from Radix Bupleuri by using volatile oil extractor; the content of sai-kosaponins in Radix Bupleuri harvested in different seasons were determined by visible spectrophotometry taking saikosaponin a as index.RESULTS:The content of volatile oil was the highest in Radix Bupleuri harvested in July but the lowest in November;the content of saikosaponins was the highest when harvested in May and the lowest in September. CONCLUSION:The method is simple and rapid,and it provides scientific basis for the establishment of the best harvesting season of Radix Bupleuri.
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Objective To establish the GC fingerprint analysis for the quality control of volatile oil in Radix Bupleuri. Methods The GC with capillary column DB-1 (30 m?0.25 mm?0.25 ?m) was used. The column was maintained at 50 ℃ for 5 min after injection then programmed at 3 ℃/min to 170 ℃ and at 5 ℃/min to 230 ℃ which was maintained for 5 min. Gasification temperature: 250 ℃, carrier gas: N_2, flow rate: 1.03 mL/min, inlet volumn: 0.5 ?L, spliting ratio: 10∶1, FID Injector temperature: 250 ℃. The internal standard was [WTBX]n-nonane used to determine 25 batches of Radix Bupleuri from different habitats by GC fingerprint. Results The 25 batches of Radix Bupleuri are classified to be the qualified and unqualified based on the results of cluster and similarity analyses. Conclusion The method is simple and reliable and it is capable of effectively controlling the quality of Radix Bupleuri.
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A one - dose 5 - 1 - 6 monoclonal antibody injected intravenously can induce atransient massive proteinuria in rat. Comparison between Radix Bupleuritreating group and PBS control group 1, 3, 5, 8 days after injection showed significant difference. Under immuno -flourescence exam, the granular precipitation along the glomerular capillaries in experimental group decreased distinctively, yet the effect of Chailing Decoction on proteinuria can not be proved by oral infusion, suggesting that Radix Bupleuri may change the permeability of glomerular capillary in rat through certain mechanism.
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In this experiment,Xiaochaihu Oral Liquid was found to possess marked functions of inhibition on hydrops of rat articulatio talocruralis(p
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AIM: Research on the change of the saponins constituents of Radix Bupleuri after being processed has been carried out. METHODS: Saikosaponin a、c、d and b2 are used as the marker ingredients; the change of saponins constituents,both after procession and saponated action,have been determined by HPLC. RESULTS: After being processed,the content of saikosaponin b2 has a significant increase,saikosaponin a,saikosaponin c,saikosaponin d and saikosaponin a + c + d all have slight decrease. After the saponated action,the content of saikosaponin a + c + d has a little change,and saikosaponin b2 has increased significantly. CONCLUSION: The change rules of saponin compounds in processed Radix Bupleuri have been revealed.