Radix Paeoniae Alba Research Articles

Studies on the Changes of Pharmacokinetics Behaviors of Phytochemicals and the Influence on Endogenous Metabolites After the Combination of Radix Bupleuri and Radix Paeoniae Alba Based on Multi-Component Pharmacokinetics and Metabolomics.

Radix Bupleuri-Radix Paeoniae Alba (RB-RPA) is a classic herb pair, which is commonly used to treat depression by soothing “liver qi stagnation” in the clinic. However, little is yet known concerning the combination mechanism of Radix Bupleuri (RB) and Radix Paeoniae Alba (RPA), their bioactive forms in vivo and the regulatory effects on the organism. The present study aimed to elucidate the changes in multi-component pharmacokinetics (PK) behavior after the combination of RB and RPA by a high-resolution full-scan mode of UPLC-HRMS method (a total of 38 components PK profiles were obtained, of which 23 components come from RB and 15 components come from RPA). Moreover, the metabolomics approach was used to analyze the dynamic response of endogenous metabolites intervened by RB-RPA, and the correlation between concentration-time curves of 38 components from RB-RPA and the dynamic response profiles of endogenous metabolites was characterized by Pearson correlation analysis. The results demonstrated that the combination of RB and RPA could significantly improve the bioavailability of five components in RB, and six components in RPA. Besides, metabolomics results indicated that a total of 21 endogenous metabolites exhibited time-dependent changes in response to the RB-RPA administration, of which 12 endogenous metabolites were significantly increased, and nine endogenous metabolites were significantly decreased. Furthermore, correlation analysis results indicated that the components with significantly improved bioavailability after combination such as saikogenin F, saikogenin G, albiflorin, methyl gallate, paeonimetabolin II were significantly positively correlated with picolinic acid, a metabolite with neuroprotective effect; saikogenin F, saikogenin G were significantly positively correlated with itaconic acid, a endogenous metabolite with anti-inflammatory activity; and albiflorin, paeonimetabolin II were significantly positively correlated with α-linolenic acid, a metabolite with strong protective actions on brain functions. These results indicated that the combination of RB and RPA can enhance each other’s neuroprotective and anti-inflammatory activities. In this study, A novel and efficient strategy has been developed to analyze the influence of the combination of RB and RPA in vivo behaviors by combining multi-component pharmacokinetics with metabolomics, which was contributed to clarifying the scientific connotation of herb–herb compatibility.

Removal of Matrix Interferences by Nano-MgO and Co-Adsorbents for Accurate Multi-Pesticide Residue Analysis in the Chinese Medicinal Herb, Paeoniae Radix Alba.

A simple, accurate, and high-throughput analytical method was developed to detect 123 pesticide residues in Chinese medicinal herb Paeoniae Radix Alba (PRA) by introducing nano-MgO as a highly efficient purification material based on quick, easy, cheap, effective, rugged, and safe (QuEChERS) design concept. Various PRA samples were extracted using 8 mL 0.5% acetic acid-acetonitrile solution and purified by a dispersive solid-phase extraction method with 30 mg nano-MgO, 40 mg primary secondary amine (PSA), and 40 mg octadecylsilane (C18) as the cleanup adsorbents, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). 70.7% of pesticides showed a weak matrix effect after the purification process, indicating that this method can give the precise quantitative analysis of trace pesticides residue. The method was systematically validated under optimal conditions in five different kinds of PRA samples; good linearity was observed in the concentration range of 0.5–250 μg/L or 1–250 μg/L. Pesticide recovery in each sample spiked at concentrations of 20, 50, and 200 μg/kg ranged from 98.0% to 111% and the mean relative standard deviation ranged from 2.72% to 5.70%. Furthermore, the method comparison with the traditional QuEChERS method suggested the feasibility, advantages, and potential application prospect of the present method for the multi-pesticide residue analysis in various PRA samples.

Cleavage rules of mass spectrometry fragments and rapid identification of chemical components of Radix Paeoniae Alba using UHPLC-Q-TOF-MS.

Radix Paeoniae Alba (RPA) presents several pharmacological effects, including analgesia, liver protection, and toxicity reduction. RPA consists mostly of monoterpenes and their glycosides, tannins, flavonoids, and organic acids, with monoterpenes being the main active pharmaceutical ingredients. To establish an effective method for rapid classification and identification of the main monoterpenes, flavonoids, and organic acids in RPA. We used ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) and data post-processing technology to rapidly classify and identify the monoterpenoids, flavonoids, and organic acids in RPA. We also summarised the diagnostic product ions and neutral losses of monoterpenoids, flavonoids, and organic acids in RPA reported in the literature. We identified 24 components, namely 18 monoterpenoids, one flavonoid, and five organic acids. In this study, we analysed the chemically active pharmaceutical ingredients and assessed the quality of RPA. In addition, we demonstrated that UHPLC-Q-TOF-MS can be used to qualitatively classify and identify the variety of chemical components of traditional Chinese medicines (TCMs) to a certain extent. Moreover, we confirmed that mass spectrometry can be used to identify the components of TCMs.

Carbon Dots from Paeoniae Radix Alba Carbonisata: Hepatoprotective Effect.

IntroductionThe charcoal processed product of Paeoniae Radix Alba (PRA), PRA Carbonisata (PRAC), has long been used for its hepatoprotective effects. However, the material basis and mechanism of action of PRAC remain unclear.AimTo explore the hepatoprotective effects of Paeoniae Radix Alba Carbonisata-derived carbon dots (PRAC-CDs).MethodsPRAC-CDs were characterized using transmission electron microscopy, high-resolution transmission electron microscopy, ultraviolet, fluorescence, Fourier transform infrared and X-ray photoelectron spectroscopy, X-ray diffraction, and high-performance liquid chromatography. The hepatoprotective effect of PRAC-CDs was evaluated and confirmed using the classic carbon tetrachloride acute liver injury model.ResultsPRAC-CDs averaged 1.0–2.4 nm in size and exhibited a quantum yield of 5.34% at a maximum excitation wavelength of 320 nm and emission at 411 nm. PRAC-CDs can reduce the ALT and AST levels of mice with carbon tetrachloride-induced acute liver injury and have a mitigating effect on the rise in TBA and TBIL. More interestingly, PRAC-CDs can significantly reduce MDA and increase SOD levels, demonstrating that PRAC-CDs can improve the body’s ability to scavenge oxygen free radicals and inhibit free radical-induced liver cell lipid peroxidation, thereby preventing liver cell damage.ConclusionThese results demonstrate the remarkable hepatoprotective effects of PRAC-CDs against carbon tetrachloride-induced acute liver injury, which provide new insights into potential biomedical and healthcare applications of CDs.

Therapeutic effect of herb-separated moxibustion at Jinsuo (GV 8)-eight-diagram points on diarrhea-type irritable bowel syndrome of liver stagnation and spleen deficiency

To observe the clinical therapeutic effect of herb-separated moxibustion at Jinsuo (GV 8)- eight-diagram points on diarrhea-type irritable bowel syndrome (IBS-D) of liver stagnation and spleen deficiency as compared with oral administration of pinaverium bromide tablets and Chinese herbal decoction, tongxieyaofang. A total of 126 patients with IBS-D of liver stagnation and spleen deficiency were randomized into a herb-separated moxibustion group (moxibustion group), a western medication group and a Chinese herbal medication group, 42 cases in each one. In the moxibustion group, the herb-separated moxibustion was applied to Jinsuo (GV 8)-eight-diagram points. The herbs in tongxieyaofang (fried rhizoma atractylodis macrocephalae, fried radix paeoniae alba, pericarpium citri reticulatae and radix saposhnikoviae) were ground into herbal paste and the paste was put on Jinsuo (GV 8)-eight-diagram points. The suspending moxibustion was exerted over the points for 40 min, once daily. In the western medication group, pinaverium bromide tablets were taken orally, 50 mg each time, three times a day. In the Chinese herbal medication group, the decoction of tongxieyaofang was taken orally, one dose a day, taking separately in two times. The duration of treatment was 8 weeks in each group. Before and after treatment, the symptom score of traditional Chinese medicine (TCM), gastrointestinal (GI) symptom score, the score of IBS symptom severity scale (IBS-SSS) and the score of IBS quality of life (IBS-QOL) scale were observed in patients of each group separately. The clinical therapeutic effect was evaluated. After treatment, the scores of TCM symptoms, GI symptom scores and IBS-SSS scores were all obviously reduced in each group (P<0.05). Each of the scores in the moxibustion group was lower than the western medication group and the Chinese herbal medication group respectively (P<0.05). After treatment, the scores of each of eight subscale structures of IBS-QOL scale, named dysphoria, interference with activity, body image, health worry, food avoidance, social reaction, sexual intercourse and relationship, were all increased obviously in each group (P<0.05). The scores of each of eight subscale structures in the moxibustion group were higher than the western medication group and the Chinese herbal medication group respectively (P<0.05). The total effective rate was 92.9% (39/42) in the moxibustion group, higher than 71.4% (30/42) in the western medication group and 73.8% (31/42) in the Chinese herbal medication group respectively (P<0.05). Herb-separated moxibustion at Jinsuo (GV 8)-eight-diagram points remarkably relieves gastrointestinal symptoms and improves the quality of life in patients of diarrhea-type irritable bowel syndrome of liver stagnation and spleen deficiency, and its clinical therapeutic effect is superior to oral administration of either pinaverium bromide tablets or tongxieyaofang.

Changes of Anti-tuberculosis Herbs Formula During Past Three Decades in Contrast to Ancient Ones.

To investigate the evolution of herbal medicine in treating tuberculosis (TB) and encourage anti-TB drug discovery and development. In this study, 477 ancient traditional Chinese medicine formulae were collected from the Dictionary of Traditional Chinese Medicine Prescriptions and 172 modern Chinese medicine formulae (from 1986 to 2016) were collected by searching 4 databases: WanFang Data Knowledge Service Platform, China National Knowledge Infrastructure Database (CNKI), China Science and Technology Journal Database (VIP) and Chinese Bio-medical Literature and Retrieval System (SinoMed) in Chinese. We restricted the search to publications in Chinese. Further data analysis was done using the Traditional Chinese Medicine Inheritance Support System version 2 Software. There were 425 herbs in the 477 ancient formulae and 257 herbs in the 172 modern formulae. Half of the top 30 herbs were shared by both modern and ancient prescriptions. They are Radix Ophiopogonis, Astragalus membranaceus, Fritillaria cirrhosa, Dried rehmannia glutinosa, Poria cocos, Angelica sinensis, Prepared rehmannia glutinosa, Platycodon Root, Radix paeoniae alba, Schisandra chinensis, Bighead atractylodes rhizome, Rhizoma anemarrhenae, Cortex lycii radicis and Radix Scutellariae. Only two groups of herbs with a high correlation coefficient were found in both modern and ancient prescriptions, the Dried rehmannia glutinosa with Radix ophiopogonis, and Radix ophiopogonis with Prepared rehmannia glutinosa. There were 9 and 15 core herb combinations in modern and ancient prescriptions, respectively, but no one was found simutaniously in both modern and ancient prescriptions. Although there were wide variations in the herb groups and herb combinations in the formulae, half of the top 30 herbs were found in both modern and ancient prescriptions. The core herb combinations in modern and ancient prescriptions could help us to improve the priscription for treatment of TB.

A comparative analysis of raw and processed Radix Paeoniae Alba samples using HPLC, chemometrics, and antioxidant activity.

Radix Paeoniae Alba (RPA) and its processed products are commonly used in traditional Chinese medicine, but the chaos phenomenon among processed products often occurs. In this study, we used multiple chemometric methods to analyze raw and six different processed products of RPA based on HPLC fingerprinting. Heat map analysis was used to assess the changes in chemical composition. Principal component analysis was used for classification, and the samples were divided into four classes: class 1 (raw, wine-processed, and vinegar-processed products), class 2 (bran-processed and soil-processed products), class 3 (stir-fried products), and class 4 (coke products). Further, the orthogonal partial least squares discriminant analysis model was used to obtain chemical markers among different classes. The antioxidant property of RPA is an important factor responsible for its pharmacological effects, and so the antioxidant activity of RPA was also investigated. We measured 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and hydroxyl radical scavenging ability, and ferric reducing antioxidant power. The total antioxidant activity follows the order: coke > stir-fried > soil-processed > bran-processed > wine-processed > raw > vinegar-processed products. These results suggest that different processing methods affect the chemical composition and antioxidant power of RPA, and thus, different products of RPA should not be mixed.

Improving imprinting effect by reducing sites embedding: Selective extraction of 1,2,3,4,6-penta-O-galloyl-β-d-glucose from Paeonia lactiflora Pall by hydrophilic molecularly imprinted polymers based on macromonomer and metal ion pivot

1,2,3,4,6-Penta-O-galloyl-β-d-glucose (β-PGG) is one of active component of polyphenol tannic acid in the rhizome of radix paeoniae alba, and isolating and purifying β-PGG from crude extract of Paeonia lactiflora Pall is rather challenge work. In this paper, to reduce the embedding of imprinted sites and increase the binding capacity, β-PGG imprinted monoliths were synthesized using a strategy of combination of hydrophilic macromonomer with metal ions as pivot. With a small side chain of oligoethylene glycol, oligo (ethyleneglycol) methyl ether methacrylate (OEG) was chosen as the hydrophilic macromonomer. 4-Vinylpyridine (4-VP) and ethyleneglycol dimethacrylate was used as functional monomer and crosslinker, respectively, and Ni2+ was adopted as metallic pivot. Polymerization parameters, including the ratio of functional monomer/template, ratios of 4-VP to Ni2+, and ratios of OEG to 4-VP on affinity of the resulting molecularly imprinted polymer (MIP) were systematically investigated. The structure and morphology of the obtained monolithic MIP was studied by field emission scanning electron microscopy and multipoint BET measurement. The greatest imprinting factor can be achieved to 68, which gave rise to excellent selectivity of the resulting MIP. When using the MIP as SPE absorbent, the mean recoveries for β-PGG were 74.1% with purity of 98.35%. The results demonstrated that the combination of metal ion pivot with hydrophilic macromonomer is effective approach to the balance of rigidity of imprinting cavities and polymer flexibility and increase the selectivity of the resulting MIP.

Study on the mechanism of two related Chinese herbs Danggui-Baishao in treating cardiovascular diseases based on network pharmacology

Objective By adopting network pharmacology to study the mechanism of the two classical Chinese herbs Danggui-Baishao in treating cardiovascular diseases. Methods By searching from TCMSP and related literature, together with the databases of TCMSP, SWISS and STITCH, potential active compounds of Danggui-Baishao were collected, while the targets for cardiovascular diseases were obtained from TTD, OMIM and DrugBank databases. Then the PPI network was screened for the major targets. The KEGG Pathway annotation analyses of major targets were performed by using the DAVID database. The ingredient-major target-key pathway network was constructed by Cytoscape. Results There were 17 compounds and 54 major targets in the ingredient-target-pathway network, as well as 10 key pathways, including inflammation-related pathway (TNF signaling pathway), pathways related to cardiovascular system (such as PI3K-Akt signaling pathway, FoxO signaling pathway, VEGF signaling pathway, Rap1 signaling pathway), prolactin signaling pathway and estrogen signaling pathway. Conclusions The study verified the characteristics of multi-components, multi-targets and integral regulation for Danggui-Baishao with the application of network pharmacology. It predicted that Dangui-Baishao couldtreat cardiovascular diseases mainly by regulating angiogenesis, inflammatory response and apoptosis. Key words: Angelica sinensis; Paeoniae radix alba; Couplet medicines; Cardiovascular diseases; Network pharmacology

Study on differentiation and features of the medical records about constipation in ancient books with doctors Wumen medical school

Objective The characteristics of prescription medication for constipation in Wumen doctors in Ming and Qing Dynasties were analyzed by data mining method, so as to provide reference for the treatment of contemporary clinical constipation. Methods The Wumen medical school doctors in Ming and Qing Dynasties treating constipation in clinical case were selected for digital acquisition and process. The association rules and complex networks analysis were used for data mining in order to explore experience of famous doctors in Wumen medical treated constipation. Results A total of 202 cases records of constipation were collected from acient TCM books by the database of ancient books and literatures of library of Nanjing University of Traditional Chinese Medicine. The results showed that the highest syndromes of 202 prescriptions was Qi stagnation and yin deficiency, followed by damp-heat, blood deficiency and liver-stomach disharmony, and the top ten medicines for internal use were angelica, almond, arborvitae, flaxseed, pinellia, cistanche, dried rehamnnia root, radix paeoniae alba, rhubarb, poria, licorice, etc. The core medicine in pairs included the Platycodon grandiflorum-almonds, safflower-peach kernel, safflower-angelica and other drugs outside, carthami-semen pruni-Peach kernel, carthami-semen pruni-Angelica, carthami-angelica-peach, peony-rhubarb-magnolia. Conclusions Most of the doctors in the Ming and Qing Dynasties believed that constipation was based on deficiency and reality, took Xinrun Tongluo as its treatment method, and paid attention to the treatment of activating blood circulation, warming yang and regulating qi and resolving phlegm, and emphasized the relationship between constipation and lung, spleen, liver and kidney. Key words: Constipation (TCM); Wumen doctor; Traditional Chinese medicine schools; Data mining; Drug features; Regularity of comparability; Medicine pharmacologic literature (TCM)

Total glucosides of paeony (TGP) extracted from Radix Paeoniae Alba exerts neuroprotective effects in MPTP-induced experimental parkinsonism by regulating the cAMP/PKA/CREB signaling pathway

Ethnopharmacological relevanceThe totally-amounted glucosides of paeony (TGP), which are made up of paeoniflorin, albiflorin, oxypaeoniflorin as well as benzoylpaeoniflorin, constitute the Baishao’ actively-working component extracted from Radix Paeonia alba employed in conventional oriental medicine aiming to treat cerebrovascular disorders, such as Parkinson's disease. However, its pharmacologic mechanism is not clear. Aim of the studyThe initial investigation was made on TGP's neuroprotective effects on PD of the mouse model based on 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) as well as the identification of potential involvement of a molecular signaling pathway. Materials and methodsThe evaluation of the behavioral damage as well as neurotoxicity in mice was made through MPTP. Spontaneous motor activity test, as well as a test of Rota-rod on mice was employed for the measurement of bradykinesia symptom. Additionally, liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) works as the determiner of the main monoamine neurotransmitters dopamine (DA) along with its metabolites 3, 4-dihydroxyphenylacetic acid (DOPAC) as well as homovanillic acid (HVA) based on mouse hippocampus connected with the anti-Parkinson's disease like effect of TGP. Besides, the measurement of the effects of TGP treatment on the expressions level of TH, DAT, a-synuclein, p-CREBS133 as well as apoptosis influence was made with the help of western-blot assay with apoptosis-related markers such as Bax and Bcl-2. ResultsThe results showed that TGP treatment lessened the behavior-based loss shown “in the spontaneous motor activity as well as the potential of falling to rotarod test”. In addition, we found that pretreatment with TGP markedly improved motor coordination, striatal dopamine and its metabolite levels. Furthermore, pretreatment of TGP conducted the protection for dopaminergic neurons with the prevented MPTP-induced reductions within the tyrosine hydroxylase (TH), substantia nigra dopaminergic transporter (DAT), as well as increasing α-synuclein protein levels with transformed dopamine catabolism as well as inhibited dopamine turnover. Besides, TGP treatment helped reversed apoptosis signaling molecules Bcl-2/Bax’ reduction; meanwhile improving p-CREBS133 the factor of growth signaling in the substantia nigra’ decrease. ConclusionThese results suggested that TGP can enhance dopaminergic neuron's cell survival in the SNpc in virtue of the activated cAMP/PKA/CREB factor of growth on inhibiting the pathway of second messenger apoptosis as well. In conclusion, the current findings indicate TGP is expected to be a new cure for PD.

Dissipation Behavior of Three Fungicides during the Industrial Processing of PaeoniaeRadix Alba and Associated Processing Factors.

Before being administered as medicinal products, Chinese herbal medicines (CHMs) must be processed and decocted for human consumption. While the presence of pesticide residues in CHMs is a major concern, pesticide dissipation behavior during CHM processing has rarely been reported. In this study, the dissipation of three pesticide residues in the CHM Paeoniae Radix Alba (PRA) was investigated during each step of industrial processing. The boiling process was found to significantly reduce pesticide residues (61–89%), and the peeling process also contributed to pesticide degradation (29–68%). The high temperature (60 °C) during the drying process led to further pesticide degradation. The processing factors of all three pesticides after each processing step were less than one, and the processing factors for the overall process were lower than 0.027, indicating that industrial processing clearly reduced the amount of pesticide residues (97.3–99.4%). The findings provide guidance for the safe use of fungicides in CHMs and can help establish maximum residue limits for PRA to reduce human exposure to pesticides.

Quality standard analysis and research for Wuji-Jianwei granules

Objective To establish the quality standard of Wuji-Jianwei granules. Methods The contents of Bupleuri Radix, Codonopsis Radix, Paeoniae Radix Alba, Corydalis Rhizoma in Wuji-Jianwei granules were identified by using the thin layer chromatography (TLC) method. The contents of peoniflorin was determined by the high performance liquid chromatography (HPLC) method. Results The TLC had strong specificity and high separation, negative control without interference. The linear range was 0.124 8-0.748 8 μg for peoniflorin (r=0.999 6). The average recovery was 94.42% (RSD=1.46%) and the content of peoniflorin was 0.725 3 mg/g. Conclusions The method is accurate and reasonable, and can be used for the quality control of Wuji-Jianwei granules. Key words: Quality control; Bupleuri radix; Codonopsis radix; Paeoniae radix alba; Corydalis rhizoma; Peoniflorin; Wuji-Jianwei granules

Chinese herb pair Paeoniae Radix Alba and Atractylodis Macrocephalae Rhizoma suppresses LPS-induced inflammatory response through inhibiting MAPK and NF-\u03baB pathway

BackgroundThe combination of Radix Paeoniae Alba (RPA) and Rhizoma Atractylodis Macrocephalae (RAM) has long been used as a classic herb pair for the treatment of gynecologic and gastrointestinal diseases, but the underlying mechanisms of the herb pair remain unknown. This study aims to explore the anti-inflammatory potentials of RPA–RAM herb pair and to elucidate the underlying mechanisms.MethodsThe bioactive parts of RPA–RAM were extracted and screened through the inhibitory effects against nitric oxide (NO) production. The effects of optimized RPA–RAM extracts (OPAE) on inflammation-associated mediators were investigated by Western blotting, real-time quantitative PCR (RT-qPCR), Enzyme-linked immunosorbent (ELISA) and immunofluorescence staining.ResultsOPAE potently suppressed the productions of NO, TNF-α, IL-6 and MCP-1 in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages, concentration-dependently inhibited protein level of inducible nitric oxide synthase (iNOS), dramatically downregulated mRNA expression of iNOS, TNF-α, IL-6 and MCP-1. In addition, OPAE significantly prevented phosphorylation and degradation of inhibitory kappa Bα (IκBα) and subsequently restrained the nuclear translocation of NF-κB p65. Pretreatment with OPAE also attenuated the LPS-induced phosphorylation of ERK, JNK and p38.ConclusionsOur findings demonstrated that OPAE suppressed inflammatory responses in LPS-stimulated RAW 264.7 macrophages by decreasing critical molecules involved in MAPK and NF-κB pathway, suggesting that the herb pair could be a promising therapeutic candidate for inflammation-related diseases.

Brain metabonomics study of the antidepressant-like effect of Xiaoyaosan on the CUMS-depression rats by 1H NMR analysis

Ethnopharmacological relevanceXiaoyaosan (XYS), a famous TCM prescription with a long history of clinical use for relieving a wide variety of depression symptoms, consists of Radix Bupleuri (Bupleurum chinense DC.), Radix Angelicae Sinensis (Angelica sinensis (Oliv.) Diels), Radix PaeoniaeAlba (Paeonia lactiflora Pall.), Rhizoma Atractylodis Macrocepha lae (Atractylodes macrocephala Koidz.), Poria (Poria cocos (Schw.)Wolf), Radix Glycyrrhizae (Glycyrrhiza uralensis Fisch.), Herba Menthae Haplocalycis (Mentha haplocalyx Briq.), and Rhizoma Zin-giberis Recens (Zingiber officinale Rosc.). Aim of the studyWe aimed to characterize the diversity and variation of two kinds metabolites of brain, i.e. aqueous and lipophilic metabolites, gaining comprehensive insights into the metabolic processes of depression-like behavior, and to reveal the mechanisms of antidepressant-like effects of XYS. Materials and methodsWe first established a CUMS (Chronic Unpredictable Mild Stress)-induced depression-like behavior model. We then extracted both aqueous and lipophilic metabolites of rat brains by a two-phase extraction method, which were subsequently characterized by two differential sequences of 1H nuclear magnetic resonance (NMR). Multivariate analysis including Principal Components Analysis (PCA) and Orthogonal Partial Least Squares-Discriminate Analysis (OPLS-DA) was applied. ResultsMetabolic profiling by PCA indicated that XYS significantly reversed the metabolic perturbation caused by CUMS. OPLS-DA showed a total of 15 metabolites including 6 lipophilic and 9 aqueous metabolites was screened as potential biomarkers involved in CUMS-induced depression-like behavior. On top of this, five pathways including (1)D-glutamine and D-glutamate metabolism, (2) valine, leucine and isoleucine biosynthesis, (3) alanine, aspartate and glutamate metabolism, (4) taurine and hypotaurine metabolism and (5) arginine and proline metabolism were recognized as the most influenced pathways associated with the process of CUMS-induced depression-like behavior. Notably, XYS significantly reversed abnormality of 5 aqueous and 4 lipophilic metabolites to normal, suggesting that XYS synergistically mediated abnormalities of multiple pathways (1), (3), (4) and (5). ConclusionsIt is the first report to investigate the antidepressant-like effects and underlying mechanisms of XYS from the perspective of brain metabolites. In a broad sense, this study brings novel and valuable insights to evaluate the efficacy of traditional Chinese medicine (TCM), to interpret mechanisms, and to provide the theoretical basis for further research on therapeutic mechanisms in clinical practice.

Radix Paeoniae Alba increases serum estrogen level and up-regulates estrogen receptor expression in uterus and vagina of immature/ovariectomized mice.

Radix Paeoniae Alba (RPA) is widely used in clinical treatment for gynecological diseases, particularly abnormal menstruation, menstrual pain, and breast tenderness; however, no scientific evidence base links RPA to estrogen replacement therapy. In this study, we characterize estrogenic activity of RPA using immature and ovariectomized (OVX) mice together with in vitro studies focus on estrogen receptor (ER) pathway for molecular mechanism. RPA treatments demonstrated significant estrogenic activity, as indicated by promoting the development of uterus and vagina in immature mice, reversing the atrophy of uterus and vagina in OVX mice, up-regulating the expressions of ERα and ERβ at protein and mRNA level in reproductive tissues. Meanwhile, RPA significantly increased serum estradiol and clearly decreased serum luteinizing hormone and follicle-stimulating hormone of immature/OVX mice. Moreover, RPA could induce ER positive MCF-7 cell from S-phase to G2 stage and induce proliferation and no influence on ER negative MDA-MB-231 cell. RPA could bind with ERα and ERβ and significantly stimulate ERα/β-estrogen response element (ERE) luciferase reporter gene expression. All activities were inhibited by the ER antagonist ICI 182,780. This study illustrates RPA exerts estrogenic effects by stimulating biosynthesis of estrogen in circulation, up-regulating ERs in target tissues, and mimicking the estrogen through ER-ERE-dependent pathway.

Profiling and identification of chemical components of Shenshao Tablet and its absorbed components in rats by comprehensive HPLC/DAD/ESI-MSn analysis

AbstractShenshao Tablet (SST), prepared from Paeoniae Radix Alba (PRA) and total ginsenoside of Ginseng Stems and Leaves (GSL), is a traditional Chinese medicine (TCM) preparation prescribed to treat coronary heart disease. However, its chemical composition and the components that can migrate into blood potentially exerting the therapeutic effects have rarely been elucidated. We developed an HPLC/DAD/ESI-MSn approach aiming to comprehensively profile and identify both the chemical components of SST and its absorbed ingredients (and metabolites) in rat plasma and urine. Chromatographic separation was performed on an Agilent Eclipse XDB C18 column using acetonitrile/0.1% formic acid as the mobile phase. MS detection was conducted in both negative and positive ESI modes to yield more structure information. Comparison with reference compounds (tR, MSn), interpretation of the fragmentation pathways, and searching of in-house database, were utilized for more reliable structure elucidation. A total of 82 components, including 21 monoterpene glycosides, four galloyl glucoses, two phenols from PRA, and 55 ginsenosides from GSL, were identified or tentatively characterized from the 70% ethanolic extract of SST. Amongst them, seven and 24 prototype compounds could be detectable in the plasma and urine samples, respectively, after oral administration of an SST extract (4 g·kg−1) in rats. No metabolites were observed in the rat samples. The findings of this work first unveiled the chemical complexity of SST and its absorbed components, which would be beneficial to understanding the therapeutic basis and quality control of SST.

Chemical markers for quality control of bran-fried sulfur-fumigated Paeoniae Radix Alba

In traditional Chinese medicine practice, crude herbs are often subjected to traditional processing (Paozhi in Chinese) for a special medicinal purpose. Bran-frying is one of processing methods for Paeoniae Radix Alba (PRA). Previous studies found that paeoniflorin and paeoniflorin sulfonate, a principle bioactive compound and its sulfur-fumigation induced characteristic sulfur-containing derivative, could be used together with sulfur dioxide residue as chemical markers for the quality control of sulfur-fumigated PRA crude material. In this paper, the feasibility of these three markers used for the quality control of bran-fried sulfur-fumigated PRA was further investigated. First, homemade samples of sulfur-fumigated PRA with different sulfur-fumigation duration (0.5–6 h) were bran-fried, and stored for 12 months. Second, the contents of sulfur dioxide residue, paeoniflorin and paeoniflorin sulfonate were dynamically quantified respectively. Third, the variation of the marker contents and their correlation during bran-frying and storage was evaluated. A validation was conducted using commercial bran-fried PRA samples. The results showed that bran-frying caused an averaged reduction of 20% in the content of sulfur dioxide residue, and during the first two months of storage the content of sulfur dioxide residue was decreased by up to 27%, then the content was tending towards stability for the subsequent ten months of storage (RSD = 3.92%). Meanwhile, paeoniflorin and paeoniflorin sulfonate were relatively stable, the contents of which were not affected by bran-frying processing and 12 months of storage. The correlations between the contents of sulfur dioxide residue and paeoniflorin/paeoniflorin sulfonate were obviously influenced by storage duration. Since sulfur dioxide residue is a safety marker, while paeoniflorin and paeoniflorin sulfonate can reflect the inner quality and the impact extent of sulfur-fumigation on the quality of bran-fried PRA respectively, these three chemicals might be used together as markers for the quality control, and consequently to ensure the safety and efficacy of bran-fried PRA.

Study on quality standard of Bushen-Tongluo granules

Objective To establish the quality standard for Bushen-Tongluo granules. Methods Drynariae rhizoma, Paeoniae radix alba, Cyathulae radix, Chuanxiongin rhizome were identified by thin layer chromatography (TLC), and the content of naringin and paeoniflorin was determined by high performance liquid chromatography(HPLC) with chromatographic column Agilent C18 (4.6 mm×250 mm, 5 μm), as the mobile phase was acetonitrile-0.1% phosphoric acid and the scan wave length were 230 and 283 nm. The column temperature was 30 ℃. The flow rate was 1.0 ml/min. Results TLC could identify Drynariae rhizoma , Paeoniae radix alba, Cyathulae radix, Chuanxiongin rhizome effectively. Under the condition, there was a good linear relationship when the content of naringin was in 6.337 0-50.695 7 μg. There is also a good linear relationship when the content of paeoniflorin was in 26.065 8-130.328 8 μg. The average recovery rate of naringin was 97.13% and RSD was 1.19%. The average recovery rate of paeoniflorin was 96.61% and RSD was 1.51%. Conclusions The established methods are simple, specific, reproducible, and sensitive, and they can be used for the quality control of Bushen-Tongluo granules. Key words: Chromatography, thin layer; Chromatography, high pressure liquid; Quality control; Bushen-Tongluo granules; Naringin; Paeoniflorin

The prolactin-release inhibitor paeoniflorin suppresses proliferation and induces apoptosis in prolactinoma cells via the mitochondria-dependent pathway.

Prolactinomas are the most prevalent functional pituitary adenomas that cause chronic pathological hyperprolactinemia. Prolactin is known to promote cell growth and inhibit apoptosis in cells. Paeoniflorin is the principal component of radix Paeoniae alba (the main ingredient in some traditional herbal formulas clinically used for hyperprolactinemia-associated disorders). Recent findings from intensive studies have suggested that paeoniflorin regulates cell proliferation and apoptosis in many cell lines. However, the effects of paeoniflorin in pituitary tumor cells remain unknown. Here the results by the Cell Counting Kit-8 and colony formation assays showed that paeoniflorin concentration-dependently decreased cell viability in both MMQ and GH3 cells and colony formation in GH3 cells, suggesting inhibition of cell proliferation by paeoniflorin. By flow cytometry, paeoniflorin was found to increase apoptotic rate in MMQ cells. Mechanistically, Western blot results revealed that paeoniflorin enhanced protein expression of cleave caspase-9 and -3, and Bax, whereas it suppressed Bcl-2 protein expression in MMQ cells. Furthermore, paeoniflorin upregulated phosphorylated p53 protein expression, but it decreased prolactin concentration and prolactin protein expression in both MMQ and GH3 cells. Thus, the present results demonstrate that paeoniflorin inhibits cell proliferation and induces the mitochondrial pathway-mediated apoptosis in prolactinoma cells. These antitumor property is associated with inhibition of prolactin secretion. Our findings may provide new insight into the mechanisms underlying improving prolactinoma-associated disorders of paeoniflorin-enriched herbs and formulas.