Inorganic Elements in Tuber of Chinese Herbal Medicine Radix Pseudostellariae from Guizhou Province

By analyzing the contents of inorganic elements in Radix Pseudostellariae from Guizhou Province, the aim was to assess the present quality of Radix Pseu- dostellariae, set limited standards of heavy metals, establish element fingerprints, and find out the characteristic elements. ICP-MS was used to measure the content of inorganic elements and map the element fingerprints. Additionally, WM/T2-2004 was applied to evaluate the quality of heavy metal elements, and the characteristic elements were determined by principal component analysis. The results showed that the contents of inorganic elements in Radix Pseudostellariae were between 0.057 and 959 mg/kg with the coefficients of variation ranging from 0.134 to 1.478, and the contents of Cd, As, Pb, and Hg were below the Standard of WM/T2-2004 in 90% of Radix Pseudostellariae. The standard limits of heavy metals in Radix Pseu- dostellariae were Cr≤6.5 mg/kg, Cu≤10 mg/kg, As≤2.0 mg /kg, Cd≤0.3 mg/kg, Hg≤0.15 mg/kg, and Pb≤4.0 mg/kg. The features of the inorganic eJements finger- prints could provide theoretical basis of identifying the quality of Radix Pseudostel- lariae and distinguishing Radix Pseudostellariae from other Chinese herbal medicines. The characteristic inorganic elements of Radix Pseudostellariae were found to be Cd, Cu, Co, Zn, Fe, Ca, Mg, and AI. Radix Pseudostellariae from Guizhou Province contained abundant inorganic elements, and the contents of heavy metals were below the evaluation criterion. The study provided a reference for the future development of the limiting values of heaw metals in Radix Pseudostellariae.

Effects of Different Concentrations of Radix pseudostellariae Fibrous Root Extracts on Serum Cytokines in Mice and Their Correlation Analysis

To investigate the effects of Radix pseudostellariae fibrous root extracts(RPFRE) on serum cytokines in normal and immunosuppressed mice and the correlation between indexes, KM male mice were randomly divided into four groups, which were given 0(control), 0.1, 0.2, 0.4 g/kg RPFRE by gavage. The levels of IL-2, IL-4, IL-6, and IFN-γ in the serum of mice were measured 14 d after gavage. On the 15day, the remaining mice in the experimental group were intraperitoneally injected with cyclophosphamide(CY), and samples were collected on the 18day to measure serum cytokines levels. The results showed that RPFRE improved the serum cytokine level in normal mice. 0.1 g/kg RPFRE significantly increased the content of IL-2and IFN-γ(P<0.05), while 0.4 g/kg RPFRE significantly increased the content of IL-4 and IL-6(P<0.05). Correlation analysis showed that serum cytokines IL-4 and IL-6 in normal mice had an extremely significant positive correlation with RPFRE concentration(P<0.01), IL-2 had a significant or extremely significant positive correlation with IL-4, IL-6, and IFN-γ(P<0.01or P<0.05), and IL-4 had an extremely significant positive correlation with IL-6(P<0.01). After CY injection, serum cytokine levels in mice decreased and immune function was inhibited. Compared with the control group, the experimental group recovered the level of cytokines to some extent. 0.1 g/kg RPFRE significantly increased the content of IL-2 in immunosuppressed mice(P<0.05), and 0.4 g/kg RPFRE significantly increased the content of IL-4, IL-6, and IFN-γ in immunosuppressed mice(P<0.05). Correlation analysis showed that the serum cytokines in immunosuppressed mice were significantly positively correlated with RPFRE concentration(P<0.01), and there was a certain dose dependence. Moreover, the correlation coefficient between IL-2 and IL-4, IL-6, and IFN-γ, IL-4 and IL-6, and IL-6 and IFN-γ reached an extremely significant level(P<0.01). To sum up, RPFRE could improve the serum cytokines level of mice, promote the secretion of cytokines, and have a certain immune protective effect. There was a significant positive correlation between the RPFRE concentration and the serum cytokines in mice, which provided basic data support for the further study of RPFRE immune function.

Analysis of the mechanism of Radix Astragali-Radix Pseudostellariae in the treatment of chronic heart failure based on network pharmacology

Objective:To explore and analyze the effective component,target and mechanism of action of Radix Astragali-Radix Pseudostellariae in the treatment of chronic heart failure(CHF)by network pharmacological research methods.Methods:With the help of TCMSP database,the effective chemical constituents and targets of Radix Astragali-Radix Pseudostellariae were screened out,and CHF-related proteins were obtained by Gene Cards and OMIM database.The target protein interaction network(PPI)was constructed on the String database platform,and using DAVID database to conduct the GO function and KEGG pathway enrichment analysis on the target of the obtained astragalus-pseudostellariae for the treatment of CHF.Results:Radix Astragali-Radix Pseudostellariae were selected as 28 effective chemical components and 104 therapeutic targets for CHF.There were obtained 49 GO functional annotations and 86 KEGG signaling pathways.Conclusion:Radix Astragali-Radix Pseudostellariae can treat CHF through multi-target and multi-pathway.

Immunoprotection Effects of Radix Pseudostellariae Fibrous Root Extraction (RPFRE) on Mice and Its Relationship with Antioxidant Function

[Objective] The paper was to explore the immunoprotection effects of Radix Pseudostellariae fibrous root extraction(RPFRE) on experi-mental mice and its relationship with antioxidant function. [Method] KM male mice were randomly divided into 4 groups based on RPFRE gradient concentration, and the immune function indexes and antioxidant indexes of mice were determined at 14 d post intragastric administration of RPFRE.The remaining mice in the experimental group were intraperitoneally injected with cyclophosphamide(CY) from the 15^(th) to 17^(th) day, and samples were collected to determine the above indexes on the 18^(th) day. [Result] Intragastric administration of RPFRE for 14 d improved the spleen index,thymus index, T/B cell stimulation index and total antioxidant capacity(T-AOC) of normal mice, and extremely increased the thymus index and TAOC(P<0.01). The T-AOC had extremely positive correlation with the spleen index, thymus index and B cell stimulation index( P<0.01), and had significantly positive correlation with the T cell stimulation index(P<0.05). From the 15^(th) to 17^(th) day of the experiment, the immune function indexes and antioxidant indexes of mice decreased and the malondialdehyde(MDA) content increased. The experimental groups of 0.1, 0.2 and 0.4 g/kg RPFRE promoted the immunoprotection and antioxidant effect of mice. RPFRE concentration had extremely positive correlation with 4 indexes of immune function and antioxidant indexes T-AOC and SOD(P<0.01), but had extremely negative correlation with MDA(P<0.01). There was a significantly positive correlation between immune function indexes and antioxidant indexes of mice after CY intervention( P<0.05 or P<0.01). [Conclusion] RPFRE can improve the immune function indexes and antioxidant indexes of normal mice and immunosuppressed mice and has immunopro-tection effect on immunosuppressed mice, and there is a significant correlation between immune function indexes and antioxidant indexes of mice.

Network Pharmacology Investigation in the Mechanism of Radix Pseudostellariae-Rhizoma Anemarrhenae Therapy for Diabetes Mellitus

Objective:To explore the targets and signaling pathways of Radix Pseudostellariae-Rhizoma Anemarrhenae treating diabetes mellitus by network pharmacology and expound its mechanism.Methods:The TCMSP and SymMap databases were used to screen out the active ingredients and targets of Radix Pseudostellariae-Rhizoma Anemarrhenae.The GenCards database was used to screen the predicted targets of diabetes mellitus.Two targets were mapped.The Cytoscape 3.7.2 software was used to construct the network diagram of Chinese medicine-active ingredient-target and screen out the core ingredient and target.The GO,KEGG pathway enrichment analysis was carried out by OmicShare and David.Results:According to the screening criteria,23 active ingredients of Radix Pseudostellariae-Rhizoma Anemarrhenae and 146 potential targets were obtained.138 common targets were obtained related with diabetes mellitus,including 5 key components and 37 core targets.The enrichment of biological process in GO is mainly related to the reaction to organic substances,the response of cells to chemical stimulation,the reaction to chemical substances,etc..The molecular function is mainly related to the enzyme binding,the same protein binding,and the activity of nuclear receptor.The cell composition is mainly associated with the membrane raft,membrane microdomain,membrane region,etc..The KEGG pathway enrichment is mainly concentrated in TNF,Toll-like receptors,NOD-like receptors,insulin resistance and type II diabetes.Conclusion:Radix Pseudostellariae-Anemarrhena asphodeloides plays the anti-inflammatory,antioxidant,hypoglycemic,and insulin-resistance improving role through multiple targets and pathways,and therefore achieves the effect on diabetes mellitus.

Study on the Mechanism of Pseudostellariae Radix in Regulating Angiogenesis Based on Network Pharmacology and a Dual-screening System

[Objectives]This study was conducted to clarify the action mechanism of Pseudostellariae Radix in regulating angiogenesis by using network pharmacology and a dual-screening system,and to provide a basis for its clinical treatment of cardiovascular diseases.[Methods]The TCMSP database was used for preliminary screening to obtain the active compounds of Pseudostellariae Radix and the protein targets of its action.GeneCards and OMIM databases were used to search for targets related to angiogenesis.Cytoscape 3.9.1 was used to construct a drug-target network and protein interaction network of Pseudostellariae Radix in angiogenesis.The GO enrichment analysis and KEGG pathway analysis of the targets of Pseudostellariae Radix in angiogenesis were carried out on Metascape platform.The effects of the screened active compounds were verified using a dual-screening system.[Results]Six active components of Pseudostellariae Radix,luteolin,acetin,beta-sitosterol,linarin,schottenol and 1-monolinolein,were screened by TCMSP database;and the six active components were predicted with 78 common target proteins related to angiogenesis,of which 19 were core targets.Pseudostellariae Radix mainly intervened in angiogenesis through domain specific binding,ubiquitin-like protein ligase binding,kinase binding and other molecular functions to regulate biological processes such as membrane microdomain,plasma membrane raft and caveola.The results of KEGG enrichment indicated that pathways in cancer,lipid and atherosclerosis,hepatitis B,apoptosis,toxoplasmosis and other key pathways might be the mechanism for the intervention of angiogenesis.The results of the dual-screening system showed that luteolin,acacetin,beta-sitosterol and linarin protected HUVECs and promoted zebrafish angiogenesis.[Conclusions]This study preliminarily demonstrated that luteolin,acacetin,beta-sitosterol and linarin could intervene in angiogenesis through multiple targets and multiple pathways,providing ideas and a scientific basis for the treatment of cardiovascular diseases.

基于Radix-4 Booth编码的并行乘法器设计

速度和面积是评价乘法器单元性能优劣的两个基本指标。针对当前乘法器设计难以平衡版图面积和传输延时的问题,采用Radix-4 Booth算法,设计了一种新型的16位有符号定点乘法器。在部分积生成过程中,首先改进对乘数的取补码电路,然后优化基数为4的改进Booth编码器和解码器,此结构采用较少的逻辑门资源,并且易对输入比特进行并行化处理。在Wallace压缩电路中,对符号扩展位进行预处理并设计新的压缩器结构,优化整个Wallace压缩模块。在第二级压缩过程中提前对高位使用纹波进位加法器结构计算,减小了多bit伪和的求和位数。在求和电路中,使用两级超前进位加法器结构,在缩短关键路径传输延时的同时避免增大芯片面积,提高了乘法器的运行速度。新型定点乘法器与已有的乘法器结构相比,减少了12.0%的面积,降低了20.5%的延时。

Effects of Radix Puerariae, Radix Rehmanniae and Their Compatibility on Blood Glucose and Blood Lipids in Mice

[Objectives]To explore the effects of the compatibility of Radix Puerariae and Radix Rehmanniae on blood glucose and blood lipids in diabetic mouses.[Methods]Diabetic mouse model was established.The body weight and fasting blood glucose of mice were measured after 7 and 14 d of administration,and the biochemical indicators of blood lipids(TC,HDL-C,and LDL-C)were detected after 14 d of administration.[Results]Compared with the Radix Puerariae group and Radix Rehmanniae group,the compatibility group(1:2)had the best hypoglycemic effect(P<0.05),and TC and LDL-C in the compatibility group(2:1)significantly decreased(P<0.05),while HDL-C in the compatibility group(1:1)significantly increased(P<0.05).[Conclusions]Radix Puerariae,Radix Rehmanniae and their combination can reduce the blood glucose of diabetic mice.The compatibility group(1:2)had a significant hypoglycemic effect(P<0.05),and LDL-C in the compatibility group(2:1)significantly declined,while HDL-C in the compatibility group(1:1)rose significantly.

Enhanced identification and localization of metabolites in Scutellariae Radix using ion mobility enabled MALDI-Q-TOF/MS imaging

Unraveling the distribution of metabolites in traditional Chinese medicine(TCM)provides direct indications for understanding their regulatory and functional basis,which is of paramount significance for better utilization and quality control of medicinal plants[1].Recently,imaging techniques such as near-infrared spectroscopy,Raman spectroscopy,and mass spectrometry(MS)were explored to reveal the spatial context of component accumulation and localization[2,3].

Puerariae Radix protects against ulcerative colitis in mice by inhibiting NLRP3 inflammasome activation

Ulcerative colitis(UC)is a common inflammatory disease of the gastrointestinal tract.Traditional Chinese medicine(TCM)has long been used in Asia as a treatment for UC and Puerariae Radix(PR)is a reliable anti-diarrheal therapy.The aims of this study were to investigate the protective effect of PR using the dextran sulfate sodium salt(DSS)-induced UC model in mice and identify molecular mechanisms of PR action.The chemical constituents of PR via ultra-performance liquid chromatography/tandem mass spectrometry and identified potential PR and UC targets using a network pharmacology(NP)approach were obtained to guide mouse experiments.A total of 180 peaks were identified from PR including 48 flavonoids,46 organic acids,14 amino acids,8 phenols,8 carbohydrates,7 alkaloids,6 coumarins and 43 other constituents.NP results showed that caspase-1 was the most dysregulated of the core genes associated with UC.A PR dose of 0.136 mg/g administered to DSS treated mice reversed weight loss and decreased colon lengths found in UC mice.PR also alleviated intestinal mucosal shedding,inflammatory cell infiltration and mucin loss.PR treatment suppressed upregulation of NOD-like receptor protein 3(NLRP3),cysteinyl aspartate-specific proteases-1(caspase-1),apoptosis-associated speck-like(ASC)and gasdermin D(GSDMD)at both the protein and m RNA expression levels.The addition of a small molecule dual-specificity phosphatase inhibitor NSC 95397 inhibited the positive effects of PR.These results indicated that PR exerts a protective effect on DSS-induced colitis by inhibiting NLRP3 inflammasome activation in mice.

Systematic investigation of Radix Salviae for treating diabetic peripheral neuropathy disease based on network Pharmacology

BACKGROUND Diabetic peripheral neuropathy(DPN)is a debilitating complication of diabetes mellitus with limited available treatment options.Radix Salviae,a traditional Chinese herb,has shown promise in treating DPN,but its therapeutic mech-anisms have not been systematically investigated.AIM Radix Salviae(Danshen in pinin),a traditional Chinese medicine(TCM),is widely used to treat DPN in China.However,the mechanism through which Radix Salviae treats DPN remains unclear.Therefore,we aimed to explore the mechanism of action of Radix Salviae against DPN using network pharmacology.METHODS The active ingredients and target genes of Radix Salviae were screened using the TCM pharmacology database and analysis platform.The genes associated with DPN were obtained from the Gene Cards and OMIM databases,a drug-com-position-target-disease network was constructed,and a protein–protein inter-action network was subsequently constructed to screen the main targets.Gene Ontology(GO)functional annotation and pathway enrichment analysis were performed via the Kyoto Encyclopedia of Genes and Genomes(KEGG)using Bioconductor.RESULTS A total of 56 effective components,108 targets and 4581 DPN-related target genes of Radix Salviae were screened.Intervention with Radix Salviae for DPN mainly involved 81 target genes.The top 30 major targets were selected for enrichment analysis of GO and KEGG pathways.CONCLUSION These results suggested that Radix Salviae could treat DPN by regulating the AGE-RAGE signaling pathway and the PI3K-Akt signaling pathway.Therefore,Danshen may affect DPN by regulating inflammation and apoptosis.

Multi-layered effects of Codonopsis Radix on the immune system

Recent research has highlighted the potential of Codonopsis Radix to modulate the immune system,making it a promising candidate for treating chronic inflammatory and cardiovascular diseases,tumors,and aging.However,because of the complex immune activities of its various components,a comprehensive understanding of Codonopsis Radix immune-regulating properties is still lacking.This knowledge gap hinders its widespread utilization in clinical practice.Therefore,this review aimed to assess the impact of Codonopsis Radix on the immune system and elucidate its underlying mechanisms.Additionally,we compared the immunomodulatory effects of different active ingredients derived from Codonopsis Radix to provide a theoretical basis for future investigations on immunomodulation.

Advances in Research of Anti-tumor Effect of Aconiti Radix

In this paper,the anti-tumor effects of Aconiti Radix were reviewed and summarized,and the clinical feasibility of Aconiti Radix as a potential anti-tumor drug was analyzed,in order to provide a useful reference for the future research and development of new anti-cancer drugs of Aconiti Radix.

Effect of Milk Processed Arnebiae Radix on Body Temperature of Fever Rats

[Objectives]To investigate the synergistic effect of Arnebiae Radix after processing.[Methods]The effects of raw Arnebiae Radix and milk processed Arnebiae Radix on hypothermia in yeast-induced febrile rats were compared.[Results]The processed and unprocessed Arnebiae Radix at high,medium and low doses all had a certain effect on inhibiting the rise of body temperature in rats.The high dose unprocessed group,the medium dose processed group and the high dose processed group had the best inhibitory effect on body temperature,the low dose processed group could delay the fever time,and the low dose unprocessed group had poor inhibitory effect on fever.[Conclusions]The prepared Arnebiae Radix has enhanced drug effect,and milk processed Arnebiae Radix can be used to replace common Arnebiae Radix to reduce the dosage of Arnebiae Radix and save Arnebiae Radix resources.

Antidepressant effects of Yuanzhi (Polygalae Radix) extract on chronic unpredictable mild stress-induced depression in rats: modulation of the NLRP3 inflammasome and NF-κB pathway

Objective To investigate the antidepressant effects of Yuanzhi(Polygalae Radix;PR)aqueous extract on chronic unpredictable mild stress(CUMS)-induced depression rat models and the underlying mechanisms.Methods A total of 40 male Sprague Dawley(SD)rats were randomly divided into control;model;low dose of PR(PR-L;0.5 g/kg);high dose of PR(PR-H;1 g/kg);and fluoxetine(10 mg/kg)groups;with 8 rats in each group.Except for the rats in control group;those in the other four groups underwent CUMS-induced depression modeling.PR and fluoxetine were administered intragastrically once daily;30 min prior to the CUMS procedure;for 14 consecu-tive days until the behavioral tests were performed.After CUMS modeling;the sucrose prefer-ence test(SPT);open field test(OFT);novelty-suppressed feeding test(NSFT);forced swim test(FST);and tail suspension test(TST)were employed to assess the pharmacological ef-fects of PR on the mitigation of depressive-like behaviors in rat models.Additionally;the en-zyme-linked immunosorbent assay(ELISA)was utilized to quantify the serum levels of tumor necrosis factor(TNF)-α;interleukin(IL)-6;and IL-1βin the rats.Western blot analysis was al-so conducted to evaluate the protein expression levels of nuclear factor kappa-B(NF-κB);in-ducible nitric oxide synthase(iNOS);cyclooxygenase-2(COX-2);nucleotide-binding oligomerization domain(NOD)-like receptor family pyrin domain containing 3(NLRP3);apoptosis-associated speck-like protein containing caspase recruitment domain(ASC);and caspase-1 in the hippocampal tissues of the rats.Immunofluorescence staining was per-formed to observe the morphological changes in ionized calcium-binding adapter molecule 1 positive(Iba-1+)cells in the dentate gyrus(DG)of rats with CUMS-induced depression.Results(i)Treatment with PR-H and fluoxetine resulted in significant enhancements in both the total distance and time the rats moved during tests(P<0.01 and P<0.05;respectively).Post-administration of PR-H and fluoxetine also led to statistically significant increase in su-crose preference among rats(P<0.05).Besides;PR-L;PR-H;and fluoxetine treatment markedly decreased the latency of ingestion(P<0.05;P<0.05;and P<0.01;respectively).As observed from the FST;PR-L;PR-H;and fluoxetine presented antidepressant effects on rats with CUMS-induced depression;leading to the reduction in time of their immobility(P<0.05;P<0.01;and P<0.01;respectively).The results of TST indicated reduced immobility time in rats receiving PR-H and fluoxetine treatment as well(P<0.01).(ii)Rats in model group showed an increase in the levels of Iba-1+microglia in their left and right brains in compari-son with control group(P<0.01).However;such increase was negated post PR treatment(P<0.01).Treatment with PR-L;PR-H;and fluoxetine considerably reduced the levels of inflam-matory factors(TNF-α;IL-1β;and IL-6;P<0.01).In addition;treatment of PR-L and PR-H ef-fectively counteracted the elevated levels of NLRP3;ASC;and caspase-1;and markedly down-regulated the expression levels of phosphorylated p65(p-p65);COX-2;and iNOS in rats’hip-pocampus(P<0.01).Conclusion Collectively;these findings indicate that PR exerts an antidepressant effect on rats with CUMS-induced depression partially through the modulation of the NLRP3 and NF-κB signaling pathways.

Potential application of Nardostachyos Radix et Rhizoma-Rhubarb for the treatment of diabetic kidney disease based on network pharmacology and cell culture experimental verification

BACKGROUND Diabetic kidney disease(DKD)is one of the serious complications of diabetes mellitus,and the existing treatments cannot meet the needs of today's patients.Traditional Chinese medicine has been validated for its efficacy in DKD after many years of clinical application.However,the specific mechanism by which it works is still unclear.Elucidating the molecular mechanism of the Nardostachyos Radix et Rhizoma-rhubarb drug pair(NRDP)for the treatment of DKD will provide a new way of thinking for the research and development of new drugs.AIM To investigate the mechanism of the NRDP in DKD by network pharmacology combined with molecular docking,and then verify the initial findings by in vitro experiments.METHODS The Traditional Chinese Medicine Systems Pharmacology(TCMSP)database was used to screen active ingredient targets of NRDP.Targets for DKD were obtained based on the Genecards,OMIM,and TTD databases.The VENNY 2.1 database was used to obtain DKD and NRDP intersection targets and their Venn diagram,and Cytoscape 3.9.0 was used to build a"drug-component-target-disease"network.The String database was used to construct protein interaction networks.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis and Gene Ontology analysis were performed based on the DAVID database.After selecting the targets and the active ingredients,Autodock software was used to perform molecular docking.In experimental validation using renal tubular epithelial cells(TCMK-1),we used the Cell Counting Kit-8 assay to detect the effect of NRDP on cell viability,with glucose solution used to mimic a hyperglycemic environment.Flow cytometry was used to detect the cell cycle progression and apoptosis.Western blot was used to detect the protein expression of STAT3,p-STAT3,BAX,BCL-2,Caspase9,and Caspase3.RESULTS A total of 10 active ingredients and 85 targets with 111 disease-related signaling pathways were obtained for NRDP.Enrichment analysis of KEGG pathways was performed to determine advanced glycation end products(AGEs)-receptor for AGEs(RAGE)signaling as the core pathway.Molecular docking showed good binding between each active ingredient and its core targets.In vitro experiments showed that NRDP inhibited the viability of TCMK-1 cells,blocked cell cycle progression in the G0/G1 phase,and reduced apoptosis in a concentrationdependent manner.Based on the results of Western blot analysis,NRDP differentially downregulated p-STAT3,BAX,Caspase3,and Caspase9 protein levels(P<0.01 or P<0.05).In addition,BAX/BCL-2 and p-STAT3/STAT3 ratios were reduced,while BCL-2 and STAT3 protein expression was upregulated(P<0.01).CONCLUSION NRDP may upregulate BCL-2 and STAT3 protein expression,and downregulate BAX,Caspase3,and Caspase9 protein expression,thus activating the AGE-RAGE signaling pathway,inhibiting the vitality of TCMK-1 cells,reducing their apoptosis.and arresting them in the G0/G1 phase to protect them from damage by high glucose.

Determination of Benzoylaconitine Compounds in the Decoctions of Aconiti Lateralis Radix Praeparata(Heishun Pieces),Trichosanthis Fructus and Their Combination

[Objectives]This study was conducted to determine the contents of benzoylmesaconine,benzoylaconitine and benzoylhypacoitine in the decoctions of Heishun pieces,Trichosanthis Fructus and their combination.[Methods]Heishun pieces,Trichosanthis Fructus and their combination were extracted for different time periods,and then grouped.HPLC was performed using an Agilent ZORBAX SB-C 18 chromatographic column(4.6 mm×250 mm,5μm)and acetonitrile-0.02 mol/L sodium dihydrogen phosphate as the mobile phase at a flow rate of 1 mL/min and a column temperature of 30℃,and the sample volume was 20μL.The detection wavelength was 230 nm.[Results]The total amounts of benzoylmesaconine,benzoylaconitine and benzoylhypacoitine in the single decoction group of Heishun pieces were all significantly different from those in the combined decoction group at corresponding time.[Conclusions]The total content of the benzoylaconitine type increased significantly after the combined decoction of Heishun pieces and Fructus Trichosanthis,which proves the scientificity of"eighteen incompatible medicaments,19 counteraction"in traditional Chinese medicine to some extent.

Protective mechanism of Paeoniae Radix Alba against chemical liver injury based on network pharmacology,molecular docking,and in vitro experiments

Objective:To explore and validate the potential targets of Paeoniae Radix Alba(P.Radix,Bai Shao)in protecting against chemical liver injury through network pharmacology,molecular docking technology,and in vitro cell experiments.Methods:Network pharmacology was used to identify the common potential targets of P.Radix and chemical liver injury.Molecular docking was used to fit the components,which were subsequently verified in vitro.A cell model of hepatic fibrosis was established by activating hepatic stellate cell(HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1.The cells were exposed to different concentrations of total glucosides of paeony(TGP),the active substance of P.Radix,and then evaluated using the cell counting kit-8 assay,enzyme-linked immunosorbent assay,and western blot.Results:Analysis through network pharmacology revealed 13 key compounds of P.Radix,and the potential targets for preventing chemical liver injury were IL-6,AKT serine/threonine kinase 1,jun protooncogene,heat shock protein 90 alpha family class A member 1(HSP90AA1),peroxisome proliferator activated receptor gamma(PPARG),PTGS2,and CASP3.Gene Ontology(GO)enrichment analysis indicated the involvement of response to drugs,membrane rafts,and peptide binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation.Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1,PTGS2,PPARG,and CASP3.Different concentrations of TGP can inhibit the expression of COL-I,COL-III,IL-6,TNF-a,IL-1β,HSP-90a,and PTGS2 while increasing the expression of PPAR-γand CASP3 in activated HSC-LX2 cells.Conclusion:P.Radix primarily can regulate targets such as HSP90AA1,PTGS2,PPARG,CASP3.TGP,the main active compound of P.Radix,protects against chemical liver injury by reducing the inflammatory response,activating apoptotic proteins,and promoting the apoptosis of activated HSCs.

Optimization of Preparative Separation and Purification of Total Flavonoids from Radix Puerariae by Macroporous Resin Method

Aim To screen the optimum macroporous resin and conditions for the isolation and purification of flavonoids from Radix Puerariae. Methods The static and dynamic adsorption/desorption methods were used, and the separation and purification process was evaluated by measuring the concentration of total flavonoid in the fractions with UV spectrophotometer. Results The SP70 macroporous resin was the most effective compared with other macroporous resins. The optimum conditions were screened, which were 0.5 g· mL^- 1 corresponding to crude drug for concentration of extract, pH 5 - 6, and appended 60 times the volume of the resin bed （BV） with the adsorption speed 2 BV·h^-1, and the volume of aq. 70% （V/V） ethanol as eluant was 5 BV with desorption speed 2 BV·h^-1. By this method, the final contents of total flavonoids exceeded 80%. Conclusion The SP70 macroporous resin is the most effective one for large-scale isolation and purification of flavonoids from Radix Pueraria, which meets industrial needs.

Analysis of principal isoflavone glycosides and aglycones in Radix astragali

Two major isoflavone glycosides [calycosin 7-O-β-D-glucopyranoside （1） and ononin （2）] and their aglycones [calycosin （3） and formononetin （4）] were simultaneously quantified with HPLC/DAD method. Two unknown compounds were identified as calycosin 7-O-β-D-glucopyranoside-6＇＂-O-malonate （U1） and formononetin 7-O-β-D-glucopymnoside-6＇＂-O-malonate （U2）, respectively, with LC/MS^n. Raw Radix astragli were shown to have higher contents of isoflavone glycosides （1, 2）, but lower contents of aglycones （3, 4） than the processed herbal materials. After being moistened with water and stored up for 24 h at 35 ℃, the glycosides and their m_alonates were almost completely transformed to their corresponding aglycones. The different contents of the isoflavone glycosides and their aglycones in raw and processed Radix astragali materials might be due to enzymolysis of the glycosides during processing.