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OBJECTIVE To establish the infrared fingerprints of Achyranthes bidentata from different producing areas ，and to conduct multivariate statistical analysis. METHODS The infrared fingerprints of 61 batches of A. bidentata samples were established by Spectrum for Window 3.02 and OMNIC 9.2 software. Taking the relative peak height of common peaks of infrared fingerprint as the variable ，the normal distribution analysis was carried out by Excel 2016 software；SPSS 22.0 software was used for cluster analysis and principal component analysis ，and the comprehensive score was calculated ；the orthogonal partial least squares-discriminant analysis was carried out by SIMCA 14.1 software，and the marker wave numbers affecting the quality of A. bidentata were screened by taking the variable importance in projection （VIP）＞1 as the standard. RESULTS The correlation coefficients of infrared spectra of 61 batches of A. bidentata samples were 0.967 2-0.997 7；there were 13 common peaks. The results of normal distribution analysis showed that the normal distribution curve of relative peak height of common peaks for A. bidentata from Henan and Hebei did not cross ，and the normal distribution curve of A. bidentata from Henan and Inner Mongolia crossed. The results of cluster analysis showed that when the distance between groups was 15，61 batches of A. bidentata samples could be clustered into 3 categories，including N 1-N12 were clustered into one category ，N13-N45 were clustered into one category，and N 46-N61 were clustered into one category. The results of principal component analysis showed that the cumulative variance contribution rate of the first three principal components was 91.121％；comprehensive score of qq.com A. bidentata （number N 40） in Jiabu village ，Jiaozuo City ， Henan Province was the highest （2.39）， and that of A.bidentata（number N 4）in Xin ’an village ，Anguo City ，Hebei Province was the lowest （－2.89）. The results of orthogonal 163.com partial least squares-discriminant analysis showed that 61 batches of A. bidentata samples were divided into three categories ，including N 1-N12 were clustered into one category ，N13-N28 were clustered into one category and N 29-N61 were clustered into one category. Seven marker wave numbers affecting the quality were selected. The corresponding wave numbers of VIP from large to small were 1 059，927，2 933，813，1 732，1 128 and 3 367 cm－1，1 732 cm－1 was the characteristic obsorption peak of saponins ，1 059，1 128，927 cm－1 were the characteristic obsorption peaks of glycosides. CONCLUSIONS Infrared fingerprint combined with normal distribution analysis ，cluster analysis ，principal component analysis and orthogonal partial least squares-discriminant analysis can be used to identify A. bidentata from different producing areas.
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OB JECTIVE To study the anti-inflammatory effect of couplet medicinals of Achyranthes bidentata -Eucommia ulmoides. METHODS Mouse macrophage RAW 264.7 were divided into blank group ，model group ，A. bidentata group（800 μg/mL），E. ulmoides group（800 μg/mL）and low- ，medium- and high- concentration groups of couplet medicinals of A. bidentata - E. ulmoides （400，800，1 600 μg/mL）. Excep for blank group and model group ，the other groups were added with corresponding drugs for 6 hours；then blank group was continued to add into the medium ，while model group was added into 10 μg/mL lipopolysaccharide （to induce the inflammatory model ）；other groups were added into corresponding drugs and 10 μ g/mL lipopolysaccharide. The levels of inflammatory factors [nitric oxide （NO），interleukin-1β（IL-1β），IL-6，tumor necrosis factor-α （TNF-α）] were detected ，and Jin ’s formula was used to evaluate the effects of A. bidentata -E. ulmoides . The expression of inducible nitric oxide synthase （iNOS），cyclooxygenase-2（COX-2），nuclear factor kappa-B （NF-κB）and inhibitor α of NF-κB （IκBα）as well as the phosphorylation of NF-κB p65，IκB kinase（IKK），p38 mitogen-activated protein kinase （p38 MAPK）， extracellular signal-regulated kinase （ERK）and c-Jun N-terminal kinase （JNK）were determined. RESULTS Compared with blank group，the level of inflammatory factors ，protein expression of iNOS and COX- 2 as well as the phosphorylation of NF-κB p65， IKK，p38 MAPK，ERK and JNK were increased significantly （P＜0.01），while the protein expression of IκBα was decreased significantly（P＜0.01）. After intervention of couplet medicinals of A. bidentata -E. ulmoides ，the level of inflammatory factors ，the expression or phosphorylation of above proteins were reversed significantly （P＜0.05 or P＜0.01），and couplet medicinals of A. bidentata-E. ulmoides had a synergistic effect. CONCLUSIONS The couplet medicinals of A. bidentata -E. ulmoides have synergistic anti-inflammatory effect on RAW 264.7 cells. Its mechanism may be related to the inhibition of NF-κB/MAPK signaling pathway related protein expression.
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Achyranthes bidentata Blume is widely used as a traditional Chinese medicine with the effects of nourishing the liver and kidneys and strengthening muscles and bones. In this work, a rapid and simple strategy was developed for characterizing phytoecdysteroids by ultra-high-performance liquid chromatography coupled with liner ion trap-Orbitrap mass spectrometry using electrospray ionization in the negative mode. As a result, 47 phytoecdysteroids were unambiguously or tentatively characterized. Among them, seven known compounds were identified according to the reference standards along with molecular formula, retention time and fragmentation patterns, while others were mostly potential new compounds. Through targeted isolation, the structures of three new compounds were determined by NMR spectra, which were consistent with LC-MS characterization. The present study provides an efficient method to deeply characterize phytoecdysteroids.
Subject(s)
Achyranthes/chemistry , Chromatography, High Pressure Liquid/methods , Gas Chromatography-Mass Spectrometry , Mass Spectrometry , Medicine, Chinese Traditional , Spectrometry, Mass, Electrospray Ionization/methodsABSTRACT
OBJECTIVE： To establish pre- column derivatization-HPLC fingerprint of polysaccharide from Achyranthes bidentata，and to determine the contents of monosaccharide components ，so as to provide reference for quality evaluation of A. bidentata decoction pieces. METHODS ：Taking 10 batches of A. bidentata decoction pieces from different manufacturers as samples，the polysaccharides were extracted by water extraction and alcohol precipitation ，hydrolyzed by trifluoroacetic acid ，and then derivatized by 1-phenyl-3-methyl-5-pyrazolone for HPLC analysis. The determination was performed on Hanbon Hedera C 18 column with column temperature of 30 ℃ at the flow rate of 1.2 mL/min. The mobile phase consisted of acetonitrile- 0.02 mol/L ammonium acetate solution （gradient elution ）. The detection wavelength was set at 250 nm，and sample size was 20 μL. HPLC fingerprint was established and similarity evaluation was performed for 10 batches of A. bidentata polysaccharide by using TCM Chromatogramic Fingerprint Similarity Evaluation System （2012A edition ）. The common peak was identified by comparing with the reference substance ，and cluster analysis was performed by using SPSS 25.0 software. The contents of identified monosaccharides were determined by HPLC. RESULTS ：The similarity of 10 batches of A. bidentata polysaccharide were all higher than 0.95. Nine common peaks were fixed and a total of 5 common peaks were identified ，namely anhydrous glucose （peak 1）， mannose（peak 2），rhamnose（peak 4），galacturonic acid （peak 5）and arabinose （peak 7）. Results of cluster analysis showed that S1 sample was classified into one category ；S2，S5，S8 and S 9 samples were clustered into one category ；S3，S4，S6，S7 and S10 samples were clustered into one category. Results of content determination showed that the contents of anhydrous glucose in 10 batches of samples ranged from 6.17 to 17.55 mg/g；those of mannose ranged from 3.31 to 7.66 mg/g；those of rhamnose ranged from 38.80 to 73.97 mg/g；those of galacturonic acid ranged from 2.49 to 8.95 mg/g；those of arabinose ranged from 11.30 to 28.58 mg/g. CONCLUSIONS ：Established pre-column derivatization HPLC fingerprints and content determination method can provide reference for quality evaluation of A. bidentata decoction pieces.
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OBJECTIVE：The establ ish the qu ality standard of Achyranthes bidentata decoction pieces and establish the fingerprint of decoction pieces of different origins. METHODS ：TLC method was used to identify A. bidentata decoction pieces. The contents of water ，total ash and ethanol-soluble extract in A. bidentata decoction pieces were determined according to the method in 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. UV spectrophotometry was used to determine the content of total steroid. The content of β-ecdysterone in A. bidentata decoction pieces was determined by HPLC. HPLC method was used to establish the fingerprint of 10 batches of A. bidentata decoction pieces. Using β-ecdysterone（No. 10 peak）as reference ，similarity evaluation was conducted by using Similarity Evaluation System for TCM Chromatographic Fingerprints （2012 edition），and the common peaks were determined. SPSS 21.0 software was used for cluster analysis and principal component analysis so as to evaluate the comprehensive quality of A. bidentata decoction pieces. RESULTS ：Results of TLC identification showed that the spots with the same color on the corresponding positions of β-ecdysterone and ginsenoside Ro control in chromatogram of test sample. The average water content of 10 batches of A. bidentata decoction pieces was 4.07％ -6.33％ . The total ash content was 5.04％ -6.43％ . The ethanol-soluble extract was 6.57％ -11.12％ . The linear range of total sterone （by β-ecdysterone） and β-ecdysterone were 0.01-0.08 mg/mL and 68.5-479.5 µg/mL （R2＞0.999）， respectively. RSDs of precision ，stability and repeatability tests were all less than 3％. The average recovery rates were 98.85％（RSD＝1.89％，n＝6）and 100.34％（RSD＝ 2.12％，n＝9），respectively. The average contents were 0.34％-0.56％ and 0.07％-0.09％，respectively. There were 24 common peaks in 10 batches of A. bidentata decoction pieces ，and the similarity was all over 0.900. Cluster analysis results showed that 10 batches of A. bidentata decoction pieces could be grouped into 4 categories，among which S 1，S3，S6 and S 10 were one category ， S2，S7 and S 8 were one category ，S4 and S 9 were one category ，and S 5 was one category. The results of principal component analysis showed that the cumulative variance contribution rate of the first four principal components was 86.774％. The order of comprehensive quality is S 8＞S5＞S9＞S4＞S3＞S7＞S6＞S10＞S2＞S1. CONCLUSIONS ：The established quality standard ， content deter mination method and HPLC fingerprint are stable and accurate ，and can be used for the quality evaluation of A. bidentata decoction pieces. Δ 基金项目：国家科技部重点研发计划（No.2018YFC1707105） ＊硕士研究生。研究方向：中药质量控制。E-mail：995354085@
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BACKGROUND: Achyranthes bidentata Bl.-Taxillus chinensis Danser is the most commonly used drug combination for clinical treatment of osteoarthritis, but its pharmacological mechanism is not clear. OBJECTIVE: To screen the main active chemical constituents of Achyranthes bidentata Bl.-Taxillus chinensis Danser based on the method of network pharmacology, collect the targets corresponding to the compounds, and build a drug-active ingredient-potential target network for treating osteoarthritis to treat bones and joints, and to systematically elucidate the pharmacological mechanism of inflammation. METHODS: The TCMSP database was used to obtain the active ingredients and targets of Achyranthes bidentata Bl.-Taxillus chinensis Danser, and the targets of osteoarthritis diseases obtained from the DisGeNET and GENE databases were intersected to screen out potential targets for the treatment of osteoarthritis by Achyranthes bidentata Bl.-Taxillus chinensis Danser. Using the String database and Cytoscape 3.6.1 software, we built a drug-active ingredient-therapeutic osteoarthritis potential target network and a protein-protein interaction network between potential targets. At the same time, the gene ontology (GO) biological function and Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway enrichment of potential targets were analyzed through the Omicshare cloud platform and DAVID database. RESULTS AND CONCLUSION: A total of 21 active ingredients and 48 potential targets for the prevention and treatment of osteoarthritis were screened from Achyranthes bidentata Bl.-Taxillus chinensis Danser. A total of 1 970 GO entries were obtained by GO enrichment analysis, of which 1 862 were biological processes, 29 were cellular components, and 79 were molecular functions (P < 0.01). KEGG pathway enrichment analysis screened out 76 potential signaling pathways for Achyranthes bidentata Bl.-Taxillus chinensis Danser to prevent osteoarthritis (P < 0.01). These findings indicate that Achyranthes bidentata Bl.-Taxillus chinensis Danser affects multiple metabolic pathways, affects different metabolic pathways that interacts and coordinates with each other, thereby preventing and curing osteoarthritis. This study tentatively explains the mechanism of the prevention and treatment of osteoarthritis by multi-component-multi-target-multi-pathway in Achyranthes bidentata Bl.-Taxillus chinensis Danser, which provides new ideas and clues for the prevention and treatment of osteoarthritis.
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OBJECTIVE: To study and compare the HPLC fingerprints of different polar parts of Achyranthes bidentata of different specifications, investigate their internal quality differences, and provide reference for the standard improvement and clinical use of the medicinal material. METHODS: The Achyranthes bidentata samples were refluxed with 75% ethanol, the extract was dissolved with 40 mL water and then,extracted in turn with petroleum ether, chloroform, ethyl acetate and n-butanol, each extracted phase and aqueous phase were obtained and concentrated. The HPLC fingerprint of each part was collected. The data was analyzed by similarity and comprehensive clustering. Meanwhile, the fingerprints of different parts were compared and analyzed. RESULTS: Eleven common peaks were determined in the HPLC fingerprints of petroleum ether fraction and chloroform extract fraction, 10,19,and 8 common peaks were determined in the HPLC fingerprints of ethyl acetate fraction, n-butyl alcohol fraction,and water extract respectively. The similarity of the fingerprints of chloroform parts was quite low.The similarity of the fingerprints of other parts was higher,all above 0.9. The difference in the fingerprints of petroleum ether parts was mainly reflected in the peak height. The chemical compositions and peak heights of ethyl acetate, chloroform and water extracts were different. Comprehensive cluster analysis could distinguish different specifications of Achyranthes bidentata. CONCLUSION: The internal qualities of different specifications of Achyranthes bidentata are different. HPLC fingerprint established in this experiment can comprehensively reflect the chemical components distribution of Achyranthes bidentata of different specifications. It provides reference for the whole quality evaluation of different specifications of Achyranthes bidentata.
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Objective: To investigate the chemical constituents in the the above-ground part of the plant Achyranthes bidentata Bl. in Amarathaceae. Methods: The raw materials were extracted by heating with 70% ethanol, and the extract was prepared by column chromatography on Diaion HP-20, MCI Gel CHP-20P, Toyopearl HW-40, Sephadex LH-20, ODS and silica gel, coupled with the semi-preparative HPLC, to isolate the chemical constituents. The isolated compounds were identified according to their MS, 1H NMR and 13C NMR data. Results: Twenty one compounds were isolated and identified: haploperoside A(1), sweroside(2), 1'-O-benzyl-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside(3), 2-ethyl-3-methyl-maleimide-N-β-D-glucopyranoside(4), 3-eth- ylidene-4-methyl-2, 5-pyrrolidinedione(5), methyl-5-hydroxy-2-pyridinecarboxylate(6), pericampylinone A(7), syringaresinol(8), episyringaresinol(9), methyl 3, 4-dihydroxybenzoate(10), 4-hydroxyphenylacetone(11), 3, 4-dihydroxyphenylacetic acid methyl ester(12), 2-(4-hydroxyphenyl)-ethanol(13), homovanillyl alcohol(14), dihydrosinapyl alcohol(15), 4-hydroxybenzoic acid (16), caffeic acid(17), 3-(4-hydroxy-3, 5-dimethoxyphenyl)propane-1, 2-diol(18), methyl(E)-3-(4-hydroxyphenyl)acrylate (19), trans-ferulic acid(20)and vanillic acid(21). Conclusion: Compounds 1-7 and 9-19 were isolated from the title plant for the first time.
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To investigate the " drug-guide" effect of Achyranthes bidentata saponins( ABS) and geniposide( GE) in the treatment on adjuvant arthritis( AA) rats. A UHPLC-MS/MS method for the quantitative determination of GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa in rat blood and joint dialysate was established. After single or combined administration with ABS and GE was given to AA rat model,a microdialysis sampling method for rat joint cavity and jugular vein blood vessels was established to collect microdialysis samples. Waters Acquity HSS C_(18) column was used to separate the above four components,with mobile phase as acetonitrile-0. 1% formic acid water as mobile phase for gradient elution. ESI source was adopted for mass spectra in a negative ion scanning mode. Multiple reaction monitoring( MRM) mode was applied to detect the above four components. The methodological results showed that GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa demonstrated a good linear relationship within the concentration ranges of 2-4 000,16-4 096,14-3 584,23-5 888 μg·L-1 respectively. The precision,accuracy,stability and matrix effect of these four ingredients reached the requirements of quantitative analysis of biological samples. The pharmacokinetic results demonstrated that the combined administration of ABS and GE( 60 mg·kg~(-1)+60 mg·kg~(-1)) can increase the degree of GE in joint cavity distribution,and the AUCjoint/AUCplasmwere twice of that of single administration of GE( 60 mg·kg~(-1)),which indicated that ABS might played a vital role in GE's distribution to joint cavity. Moreover,there was no significant difference between the distribution trend of total three ABS and GE in rats. The pharmacodynamics results showed that the combined administration of ABS and GE has stronger effects on paw swelling,arthritis index and synovial pathomorphology of AA rats than single administration of GE,which suggested that ABS might improve GE's anti-inflammatory effect in AA rats. Based on the above results,ABS has a targeting effect in increasing GE's concentration in joint cavity,with a synergy in efficacy.
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Animals , Rats , Achyranthes , Chemistry , Arthritis, Experimental , Drug Therapy , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacokinetics , Iridoids , Pharmacokinetics , Microdialysis , Reproducibility of Results , Saponins , Pharmacokinetics , Tandem Mass SpectrometryABSTRACT
OBJECTIVE： To investigate the potential effective components and mechanism of Achyranthes bidentata in the treatment of osteoporosis （OP）. METHODS： The effective components of A. bidentata were retrieved from the TCMSP database， and corresponding targets of them were collected. The targets related to OP were retrieved from DisGeNET database. TBtools 1.0 mapping software was used to draw the Wayne diagram， and screen the intersecting targets of effective components of A. bidentata and disease OP. Cytoscape 3.6.1 software and STRING database were used to construct and analyze the “drug-component- disease-target” network and protein-protein interaction （PPI） network； KEGG pathway enrichment analysis was conducted by using DAVID bioinformatics resource database. RESULTS： A total of 19 effective components were screened from A. bidentata， and there were 32 intersecting targets between effective components and disease OP. In “drug-component-disease-target” network， there were 45 nodes [1 for A. bidentata， 1 for OP， 11 for effective components （8 of the 19 effective components had no corresponding OP target）， 32 for intersecting targets] and 119 edges between nodes； quercetin， kaempferol， wogonin， baicalein and palmatine were important effective components. In PPI network， there were 31 nodes （1 of 32 intersecting targets was not associated with other proteins） and 212 edges， among which IL6， ESR1， MAPK1， IL8 and MAPK14 were the core targets of the network. There were 67 KEGG enrichment pathways， including rheumatoid arthritis， hepatitis B， Toll-like receptor signaling pathway， PI3K/Akt signaling pathway， JAK/STAT signaling pathway， NF-κB signaling pathway and so on. CONCLUSIONS： The main potential effective components of A. bidentata in the treatment of OP are quercetin， kaempferol， wogonin， baicalein and palmatine， the mechanism of which may be associated with cell differentiation and apoptosis， metabolism， inflammation reaction， etc. It has multi-component， multi-target and multi-system chara- cteristics.
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To establish a robust and accuracy molecular method to identify Achyranthis Bidentatae Radix and Cyathulae Radix formula granules. ITS sequences of Achyranthes bidentata and Cyathula officinalis were aligned, specific SNPs (single nucleotide polymorphisms) were excavated, specific primers were designed and allele-specific PCR method was established. The genomic DNA was successfully extracted from the herbal medicine and its formula granules by using an improved CTAB (cetyltrimethyl ammonium bromide) method and then performed PCR with the designed primers. The 187 bp specific band could be amplified only in the presentation of C. officinalis and its granules when use of C. officinalis specific primers, whereas the 162 bp band could be amplified only in the presentation of A. bidentata and its granules when use of A. bidentata specific primers. This method was also successfully applied in the identification of commercial formula granules.
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Achyranthes bidentata and Achyranthes aspera are saponin and steroid rich medicinal plants, used extensively for therapeutic treatments in Traditional Chinese Medicine (TCM) and Ayurveda. A. bidentata is reported to be one of the rare and extensively exploited medicinal plant species that face the issue of being endangered. Finding qualitative substitute with identical phyto-constituents contributing to similar composition and pharmacological benefits wil help in reducing the burden of exploitation of the natural habitats of such plants. In the present study, a comparative metabolite analysis of the whole drug and specific tissues isolated by laser micro-dissection (LMD) was carried out for both the selected species, by use of ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF MS). The results of the study indicate that the cortex and the medullary ray tissues are rich in their content of steroidal and saponin con-stituents such as (25S)-inokosterone-20,22-acetonide, ginsenoside Ro, bidentatoside II and achyranthoside B. Metabolite profiling of the whole tissues of both the species indicates presence of identical constituents. Thus, it is inferred that A. bidentata and A. aspera can be used as qualitative substitutes for each other.
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OBJECTIVE:To extract the feature of Achyranthes bidentata and Cyathula officinalis,and to establish image recog-nition method. METHODS:The microscopic image stitching of A. bidentata and C. officinalis was implemented by MATLAB. The color,invariant moment,stripes and the features of vascular bundle in cross section were extracted. The data was organized into da-ta matrix,and then data matrix was standardized by Zscore function;principal components were analyzed through Princomp func-tion. BP nerve network recognition mode was adopted. RESULTS:The microstructures in the micro images of the samples were kept integrated. The measured data of 27 characteristics were acquired in each group of sample. Through principal component analy-sis,the parameters of 11 main components were selected to establish BP never network. The average recognition rate of BP nerve network was 100% between 2 medicinal material relatives (n=50). CONCLUSIONS:The method can be used for micro-image auto Stitching of Chinese medicinal materials and image recognition of A. bidentata and C. officinalis.
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OBJECTIVE: To study the monthly dynamics of physical and chemical indexes in Achyranthis Bidentatae Radix(ABR, the dried root of Achyranthes bidentata Bl.) stored in simple and cool warehouses. METHODS: ABR was stored in simple and cool warehouses for 27 months. The color was observed. The water content was determined based on the drying method. The contents of β-ecdysone and 5-hydroxymethylfurfural (5-HMF) were determined by HPLC method. The accumulation of temperature difference between the simple and cool warehouses was evaluated with a relative temperature cumulation (RTC) method. The monthly dynamics of physical and chemical indexes of ABR was analyzed with RTC. RESULTS: As the extension of storage time, the ABR stored in the simple warehouse showed deeper color and harder texture, but the ABR stored in the cool warehouse still had soft texture without significant color change. The contents of β-ecdysone in ABR stored in the two warehouses both gradually decreased and dropped to lower than the limit of 0.030% ruled by China Pharmacopoeia when being stored for up to 27 months. The contents of 5-HMF of ABR stored in the two warehouses both increased and were higher for the sample in the simple warehouse than that in the cool warehouse. CONCLUSION: The concept of RTC is put forward and used to study the monthly dynamics of chemical constituents in traditional Chinese medicine during storage for the first time. The physical and chemical indexes of ABR varies during storage. Two years of storage time of ABR is suggested.
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OBJECTIVE To investigate chemical properties of a fructooligosaccharide (ABP-50-FOS)separated from Achyranthes bidentata and immune response in mice immunized H1N1 influenza vaccine. METHODS The methods of GPC,CE,IR and NMR were used to study chemical properties of ABP-50-FOS. BALB/c mice were immunized intramuscularly twice with H1N1 influenza vaccine (3 μg)plus ABP-50-FOS(200 μg)each mouse. The serum total antibody titer and its isotypes titers were analyzed by ELISA. The populations of CD4+,CD8+,CD3+and CD19+lymphocytes were deter?mined by flow cytometry. The proliferation activities of spleen T and B lymphocytes were determined with MTT method. The levels of cytokines interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),inter?leukin-4(IL-4),IL-12 and NO were measured by ELISA kits. RESULTS ABP-50-FOS was a fructooli?gosaccharide with moleculer mass 1885 u. Its bone linkages contained 1,2-and 1,6-fructose residues. ABP-50-FOS could induce high specific-IgG,IgG1,IgG2a,IgG2b and IgM titers after immunization with H1N1 influenza antigen twice(P<0.01). ABP-50-FOS significantly elevated the percentage of CD3+,CD4+ and CD8+ spleen lymphocytes and IFN-γ secretions(P<0.01)in vitro. It also stimulated peritoneal macrophage of mice and DC2.4 dendritic cells to produce TNF-αand IL-12p70 respectively (P<0.01). However,ABP-50-FOS inhibited secretions of NO in macrophage. CONCLUSION The fruc?tooligosaccharide ABP-50-FOS separated from A. bidentata can exhibit strong adjuvant activity for H1N1 influenza vaccine.
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OBJECTIVE:To study the effects of the compatibility of medicinal materials on the content of icariin in Gushuling granules formula. METHODS:The effects of the compatibility of medicinal materials on the content of icariin was investigated by orthogonal design with the compatibility of Astragalus membranaceus,Achyranthes bidentata and Concha ostreae as factors,using the content of icariin in decoction as index. The pH of mixture of Epimedium brevicornu and A. membranaceus,and the content of icariin in precipitation were determined. RESULTS:The effects of the compatibility of A. membranaceus on the content of icariin had statistical significance (P0.05);the interactions between two medicinal materials had not been found;the pH of the mixtures had no significant difference;the com-patibility of E. brevicornu and A. membranaceus produced precipitation,and the content of icariin in precipitation increased. CON-CLUSIONS:The compatibility of E. brevicornu and A. membranaceus can produce precipitation,so as to decrease the content of icariin in decoction,which is not influenced by pH value of decoction.
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OBJECTIVE: To study the active part of Achyranthes bidentata promotes maturation and regulates the functions of DC in tumor microenvironment. METHODS: The total saponins and the polysaccharides from dry powder of Achyranthes bidentata were extracted and identified. The tumor microenvironment by co-culturing breast cancer MCF-7 cells with DCs, and add polysaccharides, total saponins or total saponins with polysaccharides of Achyranthes bidentata to stimulate DCs were established. The expression of surface molecules on DCs and the killing activity of cytotoxic T lymphocytes (CTL) induced by DCs by Flow Cytometry were detected. ELISA was used to detect the levels of IL-β, TNF-α, IL-10 and IL-12 in the culture supernatant. Detect the protein expression of JAK, p-JAK, STAT and p-STAT3 in DCs by Western blot. RESULTS: In the tumor microenvironment, the active parts of Achyranthes bidentata, total saponins and ABPS, alone or combined used, can trigger JAK/STAT3 signaling pathway of DCs through TLR4 and Dectin-1 receptor and upregulate the expression of surface molecules, such as CD80, CD86 and CD40, to induce the maturation and differentiation of DCs, promote the secretion of TNF-α, IL-12, IL-10 and IL-β. CONCLUSION: As an effective adjuvant, the active parts of traditional Chinese medicine can enhance the effect of DC vaccine to correct the immune deficiency in tumor microenvironment, activate tumor-specific CTLs and then produce specific anti-tumor effects.
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The aim of this work was to study the plant regeneration and Agrobacterium-mediated transformation of Achyranthes bidentata using cotton EREBP gene. Results showed that the callus induction rate of stems from A. bidentata was the highest (100%) and bud was in approximately 70% of calli from stems. However bud differentiation rate of the callus from leaves and petioles was very low. Compared with ceftriaxone, 200mg/L cefotaxime could completely control Agrobacterium tumefaciens and had relatively less toxic action on the stems of A. bidentata. In addition, the induction rate of callus resistant to hygromycin was the highest when infected for 3 min and co-cultivated for 3 d. Six positive transgenic plants transformed with pCAMBIA1304-GhEREB2 expression vector were obtained and confirmed by PCR. The expression of target gene GhEREB2 was detected in five transgenic plants by RT-PCR. In brief, an efficient system of genetic transformation and plant regeneration was established for A. bidentata.
ABSTRACT
<p><b>OBJECTIVE</b>To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants.</p><p><b>METHODS</b>Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration.</p><p><b>RESULTS</b>Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of plants were established in the field.</p><p><b>CONCLUSIONS</b>The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A. aspera and A. bidentata. The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can be easily adopted for commercial large scale cultivation.</p>
Subject(s)
Achyranthes , Culture Media , Chemistry , Plant Roots , Plant Shoots , Plants, Medicinal , Survival AnalysisABSTRACT
OBJECTIVE To investigate the potential protective effect of Achyranthes bidentata polysaccharides (ABP) on diabetes mice induced by streptozocin. METHODS Male ICR mice were divided into normal control, diabetes model and ABP 50 and 100 mg·kg~(-1) (ip, once daily for 15 d) treatment groups. On the day before ABP administration and after ABP administration for 8 and 15 d, the blood glucose content was detected with a glucometer and intraperitoneal glucose tolerance test was also conducted. After ABP administration for 15 d, the mice were sacrificed and body weight, heart, liver, spleen and kidneys weights were measured. The serum insulin concentration was determined by radioimmunoassay kit. The serum activities of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and alkaline phosphatase (ALP), and serum concentrations of triglyceride (TG), total cholesterol (TC), calcium and phosphorus were measured by colorimetric method. Enzyme linked immunosorbent assay was employed to detect the concentrations of leptin, adiponectin, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum. RESULTS Compared with mice in normal control group, the body weight and serum insulin concentration decreased and blood glucose increased in diabetic model mice. ABP 50 and 100 mg·kg~(-1) treated mice were able to normalize glucose concentrations better following a glucose tolerance test, and the blood glucose level decreased by 27.4% and 16.3%, respectively, compared with that of diabetic model mice. The relative weights of spleen and kidneys, blood glucose level, serum TG and TC concentrations, and GOT, GPT and ALP activities in mice treated with ABP 50 mg·kg~(-1) were obviously lower than those of diabetes model mice. Serum leptin concentration was also markedly decreased near to normal level. However, serum concentrations of adiponectin, TNF-α and IL-6 were significantly increased comparing with diabetes model mice. ABP 100 mg·kg~(-1) had no obvious effect on serum TG and TC levels, and GPT and ALP activities. Its effects on the other parameters indicated above were similar to those in ABP 50 mg·kg~(-1) group. For the serum concentrations of insulin, calcium and phosphorus, no statistical difference could be observed among the different groups. CONCLUSION ABP possesses protective effect against streptozocin-induced diabetes in mice.