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Objective To investigate the antidepressant effect of Bupleuri Radix and Paeoniae Alba Radix drug pair based on the cAMP-CREB-BDNF pathway. Methods The rat depression model was established by CUMS. The contents of cAMP, p-CREB, BDNF, and PDE4 in rat hippocampal and cAMP levels in rat plasma were determined by ELISA. The expression of BDNF mRNA in hippocampus, hypothalamus, and cortex were measured by RT-PCR. Results Compared with the model group, the positive drug group and Bupleuri Radix and Paeoniae Alba Radix drug pair can reverse the cAMP content in the hippocampus and plasma and the decreased contents of CREB and BDNF in the rat hippocampus. At the same time, the positive drug group, Bupleuri Radix, and Paeoniae Alba Radix can increase the expression of BDNF mRNA in hippocampus, cortex, and hypothalamus of rats. Conclusion The Bupleuri Radix and Paeoniae Alba Radix drug pair has obviously antidepressant effect on CUMS rat model, which can achieve antidepressant effect by regulating cAMP-CREB-BDNF pathway.
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Objective: To investigate the effects of Paeoniae Alba Radix extracts (PAREs) on 5-HT3R mediated ion channels in primarily cultured hippocampal neurons of rats with depression. Methods: An animal model of depression was successfully developed and evaluated in rats. PARE was used for drug intervention. Serum in each group was collected, inactivated and then added into the primary hippocampal neurons for 24 h. The protein expression levels of 5-HT3AR and 5-HT3BR in the neurons of each group were examined by Western blotting (WB). The 5-HT3R channel current was recorded by a whole-cell patch clamp. Results: Compared with normal rats, the rats with depression had significantly reduction in total distance of the open-field test (OFT) and sucrose preference ratio (P < 0.01). The hippocampal neurons treated with serum of depressive rats had significantly increased protein expression of 5-HT3AR and 5-HT3BR (P < 0.05) and current density value (P < 0.05) compared to those treated with normal rat serum. Compared with the depressive rats, the rats treated with PARE and fluoxetine had significantly increased OFT (P < 0.05) and sucrose preference ratio (P < 0.01). The primary hippocampal neurons cultured with serum from PARE and fluoxetine-treated rats had significantly reduction in protein expression of 5-HT3AR and 5-HT3BR (P < 0.05, 0.01, 0.001) and current density value (P < 0.01). Conclusion: PARE can reduce the 5-HT3R ion channel current density in the rats with depression. This may be its central mechanism in treating depression.
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Objective: To study the effect of Xiaoyao Pills (containing Paeoniae Alba Radix) on peripheral blood and hypothalamic- pituitary-adrenocortical (HPA) axis in rats with syndrome of stagnation of liver qi and blood deficiency. Methods: The model of syndrome of stagnation of liver qi and blood deficiency was established by chronic restraint stress combined with radiation, and healthy male rats were randomly divided into four groups such as normal group, model group, Paeoniae Alba Radix group, and Xiaoyao Pills group. The rats in treatment groups and normal group were fed with medicines and pure water from day 1 of the experiment, respectively. The weight, peripheral blood, glucose consumption, and hypothalamic-pituitary-adrenal axis were measured. Results: Compared with the model group, the weight and amount of WBC of model group were decreased significantly (P < 0.001), and the amounts of ACTH, CORT, and CRH were increased significantly (P < 0.01 or 0.001). Glucose consumption test showed that the ability of model group to explore the novel environment was reduced (P < 0.01), Xiaoyao Pills group on the above indicators had a corresponding regulatory role (P < 0.001 or P < 0.01). Conclusion: The effect of Xiaoyao Pill on peripheral blood and HPA axis may be one of the mechanisms of its function of soothing liver to resolve depression and nourishing blood to invigorate the spleen.
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Objective: To validate the antidepressant effect of herb pair of Bupleuri Radix and Paeoniae Alba Radix by observing its interference effect in the chronic unpredictable mild stress (CUMS) depression model of rats. Methods: After the interference of herb pair of Bupleuri Radix and Paeoniae Alba Radix in CUMS depression model of rats, those changes of rat body weight, sugar water consumption, and the opening activity behavior were observed. Results: After the interference of herb pair of Bupleuri Radix and Paeoniae Alba Radix in CUMS depression model of rats for two weeks, the test indexes of rat body weight, sugar water consumption, and the opening activity behavior were significantly increased in low-, mid-, and high-dose herb pair of Bupleuri Radix and Paeoniae Alba Radix groups. The effect of herb pair of Bupleuri Radix and Paeoniae Alba Radix on improving the behavior of rats with depression was proved. In addition, Bupleuri Radix and Paeoniae Alba Radix could also improve the level of individual indicators in a certain degree. Conclusion: Herb pair of Bupleuri Radix and Paeoniae Alba Radix has the obvious antidepressant effect on rats and the antidepressant effect of Bupleuri Radix and Paeoniae Alba Radix separately is lower than it.
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Objective: Qualitative analysis of chemical components in Paeoniae Alba Radix extracts (PARE) in rat serum and brain. Methods: After ig administration of PARE, rat blood and brain tissues were collected. The chemical components in serum and brain were analyzed by HPLC-ESI-MSn. Results: By comparing retention time, MS and MSn spectra with data in the literature and reference standards, a total of 34 compounds including eight new metabolites were identified in serum, and three components including one new metabolite were identified in rat brain. Conclusion: This research will contribute to elucidate the pharmacodynamic material basis of antidepressant effect of PARE.
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Objective: To observe the change of single amine neurotransmitter content in brain of rats, to reveal the antidepressant mechanism of herb pair of Bupleuri Radix and Paeoniae Alba Radix. Methods: Chronic unpredictable mild stress (CUMS) depression model was successfully established on rats. Rats were randomly divided into control, model, positive drug (fluoxetine hydrochloride 15 mg/kg), low-, mid-, and high-dose of herb pair of Bupleuri Radix and Paeoniae Alba Radix (crude drug of 8, 16, and 32 g/kg, Bupleuri Radix-Paeoniae Alba Radix ratio 1∶1), low-, mid-, and high-dose of Bupleuri Radix (crude drug 8, 16, and 32 g/kg) group, and low-, mid-, and high-dose of Paeoniae Alba Radix (crude drug 8, 16, and 32 g/kg) groups, which were ig given drugs for 14 d. After the administration, the brain tissue of rats was taken out. The monoamine neurotransmitter [norepinephrine (NE), 5-hydroxytryptamine (5-HT), and 5-hydroxyindoleacetic acid, (5-HIAA)] content changes of material could be detected by high performance liquid chromatography (HPLC). Results: Compaed with the model group, the contents of NE and 5-HT increased obviously, while the content of 5-HIAA decreased obviously in the hippocampus, cortex, and hypothalamus in the mid-, and high-dose of herb pair of Bupleuri Radix and Paeoniae Alba Radix group (P<0.05, 0.01); In the low-dose herb pair of Bupleuri Radix and Paeoniae Alba Radix groups, the decrease-contents of NE and 5-HT could be reversed of and the level of 5-HIAA could be decreased (P<0.05) in a varying degrees. Conclusion: The herb pair of Bupleuri Radix and Paeoniae Alba Radix, Bupleuri Radix, or Paeoniae Alba Radix could play the role of anti-depression by regulation of the manoamine neurotransmitter in the brain of rats. The anti-depressant effect of the herb pair of Bupleuri Radix and Paeoniae Alba Radix is better than that of Bupleuri Radix or Paeoniae Alba Radix.
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To analyze and compare the chemical compositions of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix based on "component structure" theory. Thirteen batches of Moutan Cortex, 14 batches of Paeoniae Rubra Radix from different origins and 10 batches of Paeoniae Alba Radix from different origins were analyzed by HPLC-DAD method. Hierarchical cluster analysis and principal component analysis were used for analysis. The significant differences of principal component from Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix were investigated by using F test. HPLC fingerprints were established for 13 batches of Moutan Cortex, 14 batches of Paeoniae Rubra Radix and 10 batches of Paeoniae Alba Radix, and 7 glycosides and phenolic acids components were identified. Comparative study of Moutan Coetex, Paeoniae Rubra Radix and Paeoniae Alba Radix was conducted according to the results of hierarchical cluster analysis, principal component analysis and "component structure" theory. Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix have significant differences in mass fraction of major chemical components and their ratios, leading to different curative effects.
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Objective: A high performance liquid chromatographic method was established to simultaneously quantify the gallic acid, methyl gallate, albiflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, benzoylpaeoniflorin of red peony root, and white peony root. Methods: The content of six components from 32 batches of samples collected from different product areas and markets was determined and compared by means of this established method. The mobile phase comprised of acetonitrile and water containing 0.1% phosphoric acid. Flow rate was 0.8 mL/min and column temperature was 30℃. Chromatography was monitored at 230 and 270 nm. Results: The correlation coefficients between concentration and chromatographic peak area of gallic acid, methyl gallate, albiflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, and benzoylpaeoniflorin, respectively were over 0.9999 in the ranges of 0.7830-50.10, 1.094-70.00, 2.367-151.5, 7.823-500.6, 3.125-200.0, and 0.3480-22.25 μg/mL. The average recoveries of the six compounds were 102.1%, 98.88%, 99.25%, 100.4%, 104.2%, and 100.6%, respectively. Conclusion: All the contents of albiflorin, 1,2,3,4,6-O-pentagalloylglucose, gallic acid, and methyl gallate show a remarkable difference between Paeoniae Rubra Radix and Paeoniae Alba Radix. And the latter usually contains more monoterpene glycosides than the former dose except paeoniflorin. On the other hand, Paeoniae Rubra Radix, especially originating from Paeoniae veitchii always contains more polyphenols than Paeoniae Alba Radix dose.
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Objective: To establish a UPLC fingerprint method of Paeoniae Alba Radix, and provide comprehensive evaluation of their quality in different regions. Methods: The UPLC chromatographic column was Acquity UPLC® HSS T3 (100 mm × 2.1 mm, 1.8 μm). The mobile phase was acetonitrile-0.05% phosphoric acid water with gradient elution. The detection wavelength was 230 nm and column temperature was 30 ℃ with the flow rate of 0.4 mL/min. Similarity analysis, hierarchical clustering analysis, and principal component analysis were undertaken to study 23 sets of UPLC fingerprints of Paeoniae Alba Radix. Results: A specific UPLC fingerprint of Paeoniae Alba Radix was established and eight common peaks were designated. The results showed that the qualities of the 23 sets of Paeoniae Alba Radix were not stable and the samples collected from same region and different regions both had certain differences. Conclusion: UPLC fingerprint is an available and convenient method which can be used to access the quality of Paeoniae Alba Radix rapidly.
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Objective: To compare the chemical constituents of Paeoniae Alba Radix and Paeoniae Rubra Radix, and to provide a basis for quality evaluation. Methods: NMR-based metabolomic approach combined with multivariate statistical analysis was used to investigate the differential metabolites between Paeoniae Alba Radix and Paeoniae Rubra Radix. Results: Thirty-two metabolites were identified in the 1H-NMR spectra, and the multivariate statistical analysis showed that Paeoniae Alba Radix and Paeoniae Rubra Radix could be separated clearly. Paeoniae Alba Radix contained more arginine, threonine, acetic acid, aspartic acid, glutamine, GABA, citric acid, succinate, lactate, albiflorin, 6-O-galloyl albiflorin, 1, 2, 3, 4, 6-pentagalloyglucose, and gallic acid, while Paeoniae Rubra Radix contained more alanine, α-glucose, sucrose, paeoniflorin, catechin, β-sitosterol, fatty acid, and paeonol. In addition, the Pearson correlations between differential metabolites of Paeoniae Alba Radix and Paeoniae Rubra Radix also showed apparent differences. Conclusion: The results reveal the chemical differences between Paeoniae Alba Radix and Paeoniae Rubra Radix in a holistic way, and provide a scientific basis for assessing the quality of Paeoniae Alba Radix and Paeoniae Rubra Radix, as well as the correlations between the chemical constituents and pharmacological efficacy.
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Objective: To optimize the best processing technology for bran-fried Paeoniae Alba Radix (PAR). Methods: Taking the contents and extract yields of paeoniflorin, oxypaeoniflorin, albiflorin, and gallic acid in PAR as the indexes, the affecting factors on the processing technology of bran-fried PAR was investigated by orthogonal design test and multi-index comprehensive weighted mark method. Results: The best processing technology was to add 10% bran and fry for 8 min at 200°C. Conclusion: The optimal processing technology is stable, and could provide the references for the further study of PAR.
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Objective: To establish a fast detection method during the purifying process of the extracts from Paeoniae Alba Radix based on near infrared optical fiber transmission spectroscopy. Methods: The near infrared spectroscopy (NIRS) of the eluates by water and 70% ethanol which flowed through the macroporous resin D-101 was measured by NIR transmission optical fiber probe, and the method of pattern recognition was used to study the two processes. Aiming at the process of 70% ethanol elution, the contents of constituents were determined by HPLC as the reference values, and partial least square regression (PLSR) was applied to build the calibration models of paeoniflorm and albiflorin. Results: The processes of water elution and 70% ethanol elution could be accurately monitored with pattern recognition, and the changes of the solvent system could be accurately indicated. The quantitative models were applied to successfully predict the elution curves of Paeoniae Alba Radix extracts from the same batch and different batches on macroporous resin. The root mean square errors of prediction (RMSEP) calculated by PLSR models for paeoniflorm and albiflorin were 0.124, 0.172, and 0.120, 0.133, respectively, and the correlation coefficient between the values calculated by NIRS and HPLC was all greater than 0.992. Conclusion: The method is real time, efficient, and fast, and could be applied to the fast analysis for the purifying process of Chinese materia medica.