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As one kind of v-myb avian myeloblastosis viral oncogene homolog (MYB) transcription factors, R1-MYB (MYB-related) family plays an important role in plant growth and development, as well as environmental stress and hormone signal transduction. In this study, R1-MYB family genes in Rheum palmatum L. were systematically screened based on full-length transcriptome sequencing analysis. Firstly, the physicochemical, protein domain and molecular evolution characteristics of the coding proteins were analyzed. Furthermore, the tissue expression levels of R1-MYB genes were analyzed by RNA-seq. We also investigated the expression pattern of RpMYB24 in response to various hormones and abiotic stresses. The results showed that a total of 49 R1-MYB genes were identified, which mainly encoded thermally stable hydrophilic proteins. Most of the deduced proteins were predicted to locate in nucleus. Each protein had a large proportion of random curl and α helix, and also had the W-type conserved amino acids which were the signature of MYB. R1-MYB family members were distributed in five subgroups, including circadian clock associated 1 (CCA1)-like, I-box (GATAAG)-like, CAPRICE (CPC)-like, telomere repeat binding factor (TRF)-like and TATA binding protein (TBP)-like, and the number of CCA1-like was the majority. RNA-seq revealed that 49 R1-MYB genes were differentially expressed in roots, rhizomes and leaves of R. palmatum, and the expression levels of 15 and 23 genes in roots and rhizomes were higher than those in leaves, respectively. RpMYB24 transcript was induced by abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (MeJA) treatment, and could also significantly respond to injury, low temperature and high temperature stresses except drought stress. This study systematically identified the R1-MYB family genes and their molecular characteristics, better for further gene functional validation, and then provide a scientific basis for the transcriptional regulation mechanism research into rhubarb quality formation.
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OBJECTIVE To establish H PLC fingerprint of Rheum palmatum before and after steaming with wine ，and to determine the contents of 3 differential components. METHODS HPLC method was used to establish the fingerprints of 15 batches of R. palmatum （before wine-steaming ）and prepared rhubarb （after wine-steaming ）and the similarity evaluation was conducted. The chemical pattern recognition analysis was carried out by principal component analysis ，cluster analysis ，partial least squares- discriminant analysis and orthogonal partial least squares-discriminant analysis. The contents of gallic acid ，resveratrol-4′-O- glucoside and resveratrol- 4′-O-（6″-galloyl）-glucoside in 30 batches of samples were determined. RESULTS In the fingerprint study，48 common peaks were demarcated for R. palmatum and 47 for prepared rhubarb as well as 17 common peaks were identified by reference substance. Cluster analysis and principal component analysis showed that R. palmatum derived from Qinghai before and after steaming with wine could be distinguished from those from Sichuan and Gansu. The results of content determination showed that the contents of 3 differential components in R. palmatum derived from Qinghai before and after steaming with wine were higher than those from other two production areas ；the contents of gallic acid in prepared rhubarb derived from those production areas were higher than R. palmatum ；the contents of resveratrol- 4′-O-glucoside and resveratrol- 4′-O- （6″-galloyl）-glucoside in R. palmatum derived from those production areas were higher than prepared rhubarb. CONCLUSIONS Fingerprint and content determination method established in this study can quickly ，scientifically and accurately evaluate the quality of R. palmatum from different producing areas before and after wine steaming ，which provide a basis for the processing specification and quality control of R. palmatum .
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OBJECTIVE To establish the quality standard of Kuipingning gastric floating tablets. METHODS Kuipingning gastric floating tablets were prepared and investigated in terms of property ，weight difference and friability. Crydalis yanhusuo was identified qualitatively by thin layer chromatography （TLC）method. High performance liquid chromatography method was used to determine the content of total anthraquinones in Rheum palmatum ，and set the content limit of total anthraquinones. The floating performance and release degree of the preparation were investigated ，and the release kinetic process was fitted. RESULTS Kuipingning gastric floating tablets prepared in this study were gray white to gray tablets with slight smell and bitter taste ；the weight difference and friability were all in line with relevant regulations ；the established TLC method possessed strong specificity and could accurately identify C. yanhusuo . The average content of total anthraquinones in R. palmatum was 17.95 mg/tablet，and its content limit would not be less than 14.36 mg/tablet. The initial floating time of the preparation was no more than 10 s，and the holding time was more than 8 h. The release kinetics process accorded with the Retger-Peppas release model. CONCLUSIONS The method established in this study shows good reliability ，stability and feasibility ，and can effectively control the quality of Kuipingning gastric floating tablets.
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OBJECTIVE：To study the effects of couplet medicine of Rheum p almatum-Salvia miltiorrhiza on the contents of enterogenous urotoxin and intestinal barrier function in chronic renal failure （CRF）model rats. METHODS ：Totally 55 male Wistar rats were randomly divided into sham operation group （10 rats）and modeling group （45 rats）. In sham operation group ，the kidneys were isolated but not removed ；CRF model was reproduced by 5/6 nephrectomy in modeling group. After modeling （excluding 5 dead and non-modeling rats ），modeling rats were divided into model group （water），Niaoduqing granules group （2.5 g/kg），couplet medicine of R. palmatum -S. miltiorrhiza groups（6，3 g/kg，by crude drug ），with 10 rats in each group. Sham operation group and model group were given constant volume of water intragastrically. Administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 12 weeks. After last administration ，the contents of creatinine （Scr）and urea nitrogen（BUN）in serum ，the content of urinary creatinine （Ucr） in urine were determined by automatic biochemical analyzer；creatinine clearance rate （Ccr）was calculated. The contents of enterogenous urotoxin [trimethylamine-N-oxide （TMAO），indoxyl sulfate （IS）and p-cresyl sulphate （PCS）] were determined by UPLC-ESI-MS/MS. Real-time RT-PCR and immunofluorescence assay were used to detect the mRNA and protein expression of Occludin and ZO-1 in the ileum tissue. HE staining and Masson staining were used to observe the pathologi cal changes of renal tissue. The ultrastructural changes of rat colon were observed by transmission electron microscope. RESULTS ：Compared with sham operation group ，serum contents of Scr，BUN，TMAO，PCS and IS were increased significantly in model group （P＜0.01），while urine content of Ucr ，Ccr，mRNA and protein expression of Occludin and ZO- 1 in ileum tissue were decreased significantly （P＜0.01）；renal glomerulosclerosis ， renal tubules dilation and inflammatory invasion and fibrosisin the interstitium were all found ；the intestinal epithelial barrier structure of colon tissue was severely damaged. Compared with model group ，serum contents of Scr ，BUN，TMAO，PCS and IS were decreased significantly in administration groups （P＜0.05 or P＜0.01）；the mRNA and protein expression of Occludin and ZO-1 in the ileum tissue were increased significantly （except for mRNA expression of ZO- 1 in R. palmatum -S. miltiorrhiza low-dose group （P＜0.05 or P＜0.01）；the infiltration of inflammatory cells in renal interstitium ，the degree of fibrosis and the damage of intestinal epithelial barrier structure in colon tissue were reduced. CONCLUSIONS ：Couplet medicine of R. palmatum -S. miltiorrhiza can effectively protect the residual renal function of CRF model rats ，the mechanism of which may be associated with reducing the serum contents of enterogenous urotoxin ，up-regulating mRNA and protein expresssion of Occludin and ZO- 1 in the ileum tissue so as to improve intestinal barrier function.
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MYB transcription factors play many important regulatory roles in plant growth and development, secondary metabolism, and stress adaptation processes. In this work, an MYB gene containing a complete open reading frame (ORF) was selected from the transcriptome database of R. palmatum L. RpMYB4 ORF and cloned, encoding a polypeptide of 245 amino acids with a molecular weight of 26.99 kDa. RpMYB4 lacks a signal peptide or transmembrane domain but contains two conserved DNA binding domains (HTH-MYB) of the R2R3-MYB subfamily at the N-terminus. Multiple-sequence alignment demonstrated that RpMYB4 shared as high as 61% identity with many MYB proteins from other species. Phylogenetic analysis showed that RpMYB4 had the closest relationship with FtMYB8 and was clustered in the S4 subfamily. Subcellular localization by confocal microscopy showed that an RpMYB4-GFP-fusion protein localized to the nucleus in tobacco. Real-time fluorescence quantitative PCR analyses revealed that RpMYB4 was differentially expressed in various tissues, with the highest expression in leaves, followed by petioles, rhizome, and roots, and with the lowest level in mature seeds. After treatment of R. palmatum L. seedlings with 200 μmol·L-1 MeJA, the expression of RpMYB4 in leaves was down-regulated within 24 h, and significantly up-regulated after 200 μmol·L-1 SA treatment at 12 h and 24 h. However, gene expression did not change with 200 μmol·L-1 ABA treatment. The transcripts of RpMYB4 under drought, high temperature, and mechanical injury stresses reached a peak at 24 h, 24 h, and at 3 h, respectively, while RpMYB4 expression was inhibited by low temperature stress, reaching its lowest value at 6 h. The gene showed no significant response to salt stress. Overall, RpMYB4 was cloned from R. palmatum L. for the first time, showed high expression in leaves, and was responsive to SA and various abiotic stress treatments including drought, high temperature, and mechanical injury. The results will be useful for further analysis of secondary metabolism and stress adaptations in R. palmatum L.
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Objective: To study the chemical constituents of tannin from the roots of Rheum palmatum. Methods: Chemical constituents of tannins were isolated by silica gel column chromatography, medium pressure column chromatography, and semi-preparative HPLC from the roots of R. palmatum. The structures were elucidated on the basis of spectral data analysis. Results: Four tannins, 3-O-cinnamoyl-1-O-galloyl-β-D-glucopyranoside (1), 2-O-cinnamoyl-1-O-galloyl-β-D-glucopyranoside (2), 2-O-cinnamoyl-1,6-di-O-galloyl-β-D-glucopyranoside (3), and 6-O-cinnamoyl-1-O-galloyl-β-D-glucopyranoside (4) were isolated from the roots of R. palmatum. Conclusion: Compound 1 is a new compound named palmatoside.
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Objective: To study the seeds shapes and germination characteristics, construct aseptic seedlings and callus cultural system, and provide a basis for the rapid propagation and secondary metabolic regulation of Rheum palmatum. Methods: Ten batches of R. palmatum seeds were subjected to characteristic analyses from the aspects of size, purification, weights per thousand seeds, seeds vigor, germination rates, and germination energy. The optimum disinfection system for aseptic seedlings and the optimum hormone ratio for inducing callus were screened by orthogonal test. The content of ten active components in aseptic seedlings and calli were primarily evaluated by HPLC analysis. Results: There was no significant difference in the appearances of the ten batches of R. palmatum seeds from different regions. The content of moisture, seeds vigor, germination rates, and germination energy differed apparently. The germination characteristics in Hezheng and Weiyuan counties were the best. The best disinfection group for aseptic seedlings was the combination of 75% ethanol for 30 s with 10% hydrogen peroxide for 15 min. The optimum hormones for callus induction were 6-BA (1.0 mg/L) + KT (2.0 mg/L) + NAA (1.5 mg/L). Seeds treated with different disinfectants had no significant effect on the content of ten components in germinated aseptic seedlings (P > 0.05). Seven active components were detected in callus, which was significantly lower than that in aseptic seedlings. And the content of chrysophanol-8-O-glucoside was the highest in the callus. Conclusion: The seed characteristics from Hezheng and Weiyuan counties in Gansu Province were excellent by analyses of weights per thousand seeds, seeds vigor, germination rates, and germination energy. The aseptic seedlings and callus cultural system of R. palmatum were successfully established, laying a solid foundation for further study.
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Objective:To study the quality grading standards of Rheum palmatum seedlings. Method:The quality indicators,such as fresh weight,root length,root diameter,number of lateral roots and seedling lesions,of 80 R. palmatum seedlings from different producing areas in Gansu Provincial Real Estate District were detected,and the K-mean cluster analysis results of each data were used as reference basis for dividing the seedling quality level. Result:The fresh leaf weight,root length and root diameter were used as the main grading indicators to classify the rhubarb seedlings into four grades. Specifically,the extra-large seedlings have a plant weight >30.0 g,the root diameter>2.3 cm,and the root length >30.0 cm,with lateral roots,buds intact,no lesion,wounds. The first grade seedlings have a plant weight ≥ 25.0 g,root diameter ≥ 2.1 cm,root length ≥ 25.0 cm,with no lateral root of more than 3 mm in diameter,bud intact,disease-free spots and wounds. The second grade seedlings have a plant weight ≥ 20.0 g,roots ≥ 1.8 cm,root length ≥ 20.0 cm,with no lateral roots with a basal diameter of more than 4 mm,complete buds,no lesion,wounds. The third grade seedlings have a plant weight ≥ 15.0 g,root thickness ≥ 1.5 cm,root length ≥ 15.0 cm,with a few lateral roots,intact buds,no lesion,wounds. Conclusion:Among the 80 samples of R. palmatum seedlings collected,the second and third grade seedlings accounted for 68.8% . The different grades of R. palmatum seedlings were transplanted as herbs after field processing. In terms of the fresh yield,the first grade > extra-large seedling > the second grade > the third grade seedlings, and in the plant twitch rate,extra-large seedlings > the first grade > the second grade > the third grade seedlings. Therefore,the first and second seedlings are recommended standardized production,rather than extra-large seedlings.
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OBJECTIVE： To establish a method for simultaneous determination of 8 non-anthraquinone constituents in Rheum palmatum. METHODS： HPLC method was adopted. The determination was performed on Symmetry C18 column with mobile phase consisted of methanol-0.1％ phosphoric acid （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ and detection wavelength was 280 nm. Sample size was 30 μL. RESULTS： The linear range of gallic acid， catechin， epicatechin， resveratrol 4′-O-glucopyranoside， epicatechin gallate， resveratrol 4′-O-β-D-（6″-O-galloyl）-glucopyranoside， sennoside A， 4′-hydroxyphenyl-2-butanone-4′-O-β-D-（2″-O-galloyl-6″-O-p-hydroxy cinnamyl）-glucopyranoside were 6.16-2 464 ng（r＝0.999 9）， 37.4-14 960 ng（r＝0.999 9）， 7.635-3 054 ng（r＝0.999 7）， 7.63-3 052 ng（r＝0.999 9）， 8.32-3 328 ng（r＝0.999 9）， 11.5-4 600 ng（r＝0.999 9）， 16.08-6 432 ng（r＝0.999 9）， 29.3-11 720 ng（r＝0.999 9）， respectively. The limits of quantitation were 3.48， 4.30， 6.40， 4.40， 3.39， 2.87， 8.40 and 4.95 ng， respectively. The limits of detection were 2.32， 2.58， 2.40， 2.64， 2.26， 1.23， 4.20， 2.97 ng， respectively. RSDs of precision， stability and reproducibility tests were all lower than 5％. Recoveries were 94.32％- 100.54％（RSD＝2.78％，n＝6）， 91.15％-99.36％（RSD＝3.72％，n＝6）， 92.16％-98.04％（RSD＝2.39％，n＝6）， 93.41％-100.73％（RSD＝3.17％，n＝6）， 93.89％-98.40％（RSD＝1.99％，n＝6）， 92.61％-101.74％（RSD＝3.71％，n＝6）， 92.66％-103.40％（RSD＝3.76％，n＝6）， 95.45％-102.70％（RSD＝3.06％，n＝6）， respectively. CONCLUSIONS： The established method is simple， accurate and specific， and can be used for the simultaneous determination of 8 non-anthraquinone constituents in R. palmatum.
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OBJECTIVE： To establish a method for qualitative screening and quantitative determination of indicative composition rhaponiticin from counterfeit Rheum palmatum in Compound gentian and sodium bicarbonate tablets. METHODS： Totally 45 batches of Compound gentian and sodium bicarbonate tablets were collected from 8 domestic pharmaceutical manufacurers （No. A-H） in the field of drug distribution. TLC method was used to identify rhaponiticin in the samples primarily. The content of rhaponiticin was determined by HPLC， and then UPLC-MS/MS method was used to confirm the structure of rhaponiticin. RESULTS： TLC results showed that bright blue fluorescent spots of rhaponiticin could be seen in 10 batches of samples from manufacturer D at 365 nm wavelength of ultraviolet lamp. Results of HPLC methodology investigation showed that the linear range of rhaponiticin was 0.884-88.4 μg/mL（r＝0.999 9）； the detection limit and quantitative limit were 0.707 2， 3.536 ng； RSDs of precision， reproducibility and stability tests were all lower than 1％； average recovery was 96.55％ （RSD＝0.53％，n＝6）. The contents of rhaponiticin in 10 batches of samples from manufacturer D were 0.732 4-2.890 8 mg/g. Results of UPLC-MS/MS method showed that quasimolecular ions with m/z of 419.0 and fragment ions with m/z 257.1 and 241.2 were found in both samples from manufacturer D and rhaponiticin control. CONCLUSIONS： TLC for primary screening， HPLC for content determination and UPLC-MS/MS for structure confirmation is simple， sensitive and reliable， and can be used for qualitative screening and quantitative determination of rhaponiticin in Compound gentian and sodium bicarbonate tablets. Among 45 batches of samples tested， rhaponiticin is detected in 10 batches of samples from one manufacturer， suggesting that the manufacturer substitute fake R. palmatum for genuine ones in the production of Compound gentian sodium bicarbonate tablets.
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The herbal formula, DF-02, consisting of Ephedra intermedia and Rheum palmatum are used for the treatment of the metabolic diseases such as obesity and liver fibrosis in Korean local clinics. We aimed to develop the simultaneous analytical conditions for four standards, (+)-pseudoephedrine (PSEP) and (−)-ephedrine (EP) for E. intermedia, and aloe-emodin (AE) and chrysophanol (CP) for R. palmatum using HPLC-UV techniques. The validated conditions yielded the high precision (relative standard deviation (RSD) 0.9994). As a result, four standards of DF-02 were simultaneously determined under the developed method, which will be utilized for the quality control or evaluation of DF-02 and many herbal preparations containing E. intermedia and R. palmatum.
Sujets)
Calibrage , Chromatographie en phase liquide à haute performance , Ephedra , Cirrhose du foie , Maladies métaboliques , Méthodes , Obésité , Préparations à base de plantes , Contrôle de qualité , RheumRésumé
Anthraquinones are not only the main active constituents but also the index components for the quality control of Rhei Radix et Rhizoma. To study the anthraquinone biosynthesis, Rheum palmatum L. seedlings were subjected to a high-throughput transcriptomic sequencing analysis by Illumina HiSeqTM 2000 150PE. The Illumina sequencing generated a total of 11.04 G clean data resulting in 736 309 74 clean reads, deposited in the sequence read archive (SRA accession SRP160030). Trinity do novo assembly yielded 93 646 unigenes, with an average of 1 108 nt. Functional annotation revealed that all unigenes were successfully annotated in the NR, NT, Swiss-port, PFAM, and KOG databases. GO enrichments showed that 57 subgroups were involved in biological process, cellular component, and molecular function. KEGG analysis indicated that 1 107 unigenes were implicated in 19 standard secondary metabolic pathways. 172 unigenes were analyzed to encode 28 key enzymes during the MVA, MEP, shikimic acid, and polyketide pathways related to anthraquinone biosynthesis. 125 CYP450 and 73 UGTs unigenes were related the modification of secondary metabolites in R. palmatum L. Furthermore, seven unigenes with full length cDNAs were successfully verified by RT-PCR and sequencing analyses. Then, MISA prediction produced a number of 18 885 simple sequence repeats (SSRs). Herein, the transcriptomic gene expression profiles of R. palmatum L. and candidate genes during the anthraquinone biosynthesis pathway were obtained for the first time. The results provided basic information for subsequent gene function characterization, secondary metabolic pathway analysis, and anthraquinone biosynthesis and regulation elucidation in R. palmatum L.
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OBJECTIVE:To optimize the wet paper roasting technology of Rheum palmatum. METHODS:Weighted comprehensive score composed of external properties of R. palmatum roasting with wet paper,the content of extract,total content of anthraquinone,free anthraquinones,conjugated anthraquinone and sennoside A+sennoside B,which were used as investigation indexes. L9(34)orthogonal test was adopted to investigate the three factors as the amount of paper,roasting time and roasting temperature,and optimized wet paper roasting technology of R. palmatum. Validation test was conducted. RESULTS:The optimal roasting technology was as follows as 100 g R. palmatum covered with 12 g paper,processing at 120 ℃,for 2 h. The results of validation test and pilot test showed the average comprehensive scores of roasted R. palmatum were 84.99(RSD<2.0%,n=3) and 85.06(RSD<2.5%,n=3),respectively. CONCLUSIONS:The optimal processing technology is simple and convenient with good reproducibility and operability.It lays a technical foundation for the standardized preparation of stewed R.palmatum.
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Objective To compare the effects of different extracts of Rheum palmatum on weight and tissue structure of hypothalamusand and pituitary of adult female rats,and screen the main reproductive toxicity extract.Methods The water,chloroform,ethyl acetate,and n-butanol extracts and water-soluble substance of R.palmatum were prepared by polarity gradient extraction method.Female adult rats were randomly divided into blank control group,rhubarb water extract group,and different extracts groups.The dosage of all the groups was equivalent to 4.00 g/kg crude rhubarb.Rats were administered with extracts by gavage for 60 d.Body mass growth rate of rats were calculated before and after administration.The pathological changes of hypothalamic arcuate nucleus neurons and pituitary gonadal cell were observed with light microscope.Results Compared with the blank control group,the body mass growth rate of rhubarb water extract group was decreased (P ＜ 0.05),while those in the different extracts groups were increased (P ＜ 0.01);The hypothalamic arcuate nucleus neurons of rhubarb water extract group showed chromatin marginalization,nissl substance dissolving,fuzzy boundary of nuclear membrane,as well as hell cells,and the total number of adenohypophysis cells reduced and the cells arranged in irregular.However,there were no apparent pathologic changes in different extracts groups.Conclusion Rhubarb water extract administration by long-term dose can reduce weight growth rate and result in pathologic changes of hypothalamic arcuate nucleus and adenohypophysis,while the different extracts can increase weight growth rate significantly and have little effects on the organizational structure ofhypothalamic arcuate nucleus and adenohypophysis.
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Objective: To analyze the main components in different parts of Rheum palmatum. Methods: The anthraquinones, soluble polysaccharides, cellulose, and mineral elements in the taproots, root heads, fibrous roots, root barks, petioles and leaves were detected by HPLC, UV, Weende, and ICP-AES. Results: The contents of aloe-emodin, rhein, emodin, chrysophanol, and physcion in taproots, root heads, fibrous roots, and root barks were 3.22-4.33, 1.33-2.32, 3.21-3.68, 3.22-3.76 mg/g, 0.77-1.36, 2.46-2.52, 1.16-1.46, 1.02-1.21 mg/g, 0.27-0.39, 0.28-0.34, 0.30-0.42, 0.31-0.67 mg/g, 2.85-3.70, 2.78-3.01, 4.02-4.81, 4.05-4.72 mg/g and 1.88-2.44, 1.82-2.01, 2.48-3.02, 3.61-4.46 mg/g, respectively. The contents of aloe-emodin, rhein, and emodin in leaves were 0.56-1.07, 0.45-0.69, 1.41-1.91 mg/g. The soluble polysaccharides in the taproots, petioles and leaves were 9.76%-10.42%, 5.76%-7.63%, and 3.50%-5.72%. The cellulose contents in petioles and leaves were 15.54% and 10.20%. Ca was the most abundant with 88.53 mg/g in leaves, followed by K with 32.42 mg/g, Mg with 12.93 mg/g, Al with 1.22 mg/g, and Fe with 1.17 mg/g. In the petioles, Ca with 80.60 mg/g and K with 28.73 mg/g were higher than those in roots with 21.08 and 14.09 mg/g. Na with 2.66 mg/g was also higher than that in roots with 0.26 mg/g and in leaves with 0.57 mg/g. Conclusion: The types and contents of anthraquinones in roots are higher than those in petioles and leaves, with the understanding of traditional medicinal parts. Emodin in leaves is five times as those in roots, petioles, and leaves, and also contains a certain amount of cellulose and soluble polysaccharide component, a wide variety of elements. From above analysis, the petioles and leaves could be deeper utilized.
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Diagnostic ions filter method was used to rapidly detect and identify the phenolic compounds in Rheum palmatum based on ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MSE). The representative authentic standards of phenolic compounds, including gallic acid, (+)-catechin, (－)-epicatechin, (－)-epicatechin-3-O-gallate and procyanidin B2, were subjected to analysis by UPLC-Q-TOF/MSE system with negative ion mode. Fragmentation patterns of each standard were summarized based on assigned fragment ions. The prominent product ions were selected as diagnostic ions. Subsequently, diagnostic ions filter was employed to rapidly recognize analogous skeletons. Combined with retention time, accurate mass, characteristic fragments and previous literature data, the structures of the filtered compounds were identified or tentatively characterized. A total 63 phenolic compounds (36 phenolic acid derivatives, 8 flavonoid derivatives and 19 tennis derivatives) in R. palmatum were identified, including 6 potential new compounds. The method of diagnostic ions filter could rapidly detect and identify phenolic compounds in R. palmatum This study provides a method for rapid detection of phenolic compounds in R. palmatum and is expected to complete the material basis of rhubarb.
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OBJECTIVE:To study the influential factors related to efficacy of Peach kernel-Rheum palmatum couplet medi-cines in TCM formula,and to reveal the general regularity of compatibility environment,common ratio,processing variety and dosage forms of P. kernel-R. palmatum couplet medicines. METHODS:Using Chinese Medical Prescription Selected Dictionary ed-ited by Peng Huairen as data source,142 formulas of P. kernel-R. palmatum couplet medicines were collected. By establishing data-base,compatibility types of P. kernel-R. palmatum couplet medicines,as well as common ratio,processed prodact,dosage form were classified statistically. The influential factors related to efficacy of P. kernel-R. palmatum couplet medicines with different pro-portions were summarized. RESULTS:The efficacy of P. kernel-R. palmatum couplet medicines could be divided into 6 aspects and 11 roles,including activating blood circulation to dissipate blood stasis(activating blood to relieve pain,promoting blood circula-tion to eliminate disease,activating blood to promote menstruation,breaking stagnant and eliminating blood stasis),eliminating carbuncle and detoxicating(cleaning intestine and clearing away the pathogenic heat of lung,eliminating carbuncle and expelling pus,eliminating sore and detoxicating),expelling the pathogenic heat to loosen the bowels,warming yang for dispelling cold,forti-fying the spleen and nourishing the stomach,relaxing tendon and activating blood. The compatibility environment of P. kernel-R. pal-matum couplet medicines were mainly compatible with TCM for activating qi to eliminate stasis,activating blood to promote menstru-ation,breaking stagnant and eliminating blood stasis,expelling the pathogenic heat to expel stasis. The ratio of P. kernel to R. palma-tum ranged 1 : 8-4 : 1,and the ratio ranged 1 : 8-3 : 1 when performing the role of actirating blood circalation to dissipate blood stasis. Common processed products were crude P. kernel and prepared R. palmatum. Common dosage forms were mainly decoction,pill and powder. CONCLUSIONS:Compatibility environment,ratio,processing varieties,dosage forms influence the effects of P. kernel-R. palmatum couplet medicines,especially compatibility environment.
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Objective To compare the quality of Rheum palmatum L. in different areas and ages in Gansu Province by comparing five components in Rheum palmatum L. through fingerprints and QAMS (quantitative analysis of multi-components) methods; To determine the most suitable growing areas and the best development areas in Gansu Province. Methods The water content, total ashes, and water-soluble extract content in 15 batches of Rheum palmatum L. from different areas and ages in Gansu Province were determined according to the approaches listed in the Chinese Pharmacopeia. The contents of five index components were determined through HPLC and the quality of 15 batches of Rheum palmatum L were evaluated by fingerprint and QAMS method. Results The results from 15 batches of Rheum palmatum L. showed that the water content was less than 15％, total ashes less than 10％, and water-soluble extract content less than 25％, conforming to Pharmacopoeia. There were 23 common peaks in the fingerprints of 15 batches of Rheum palmatum L.. Five of them were identified as aloe-emodin, rhein, emodin, chrysophanol and physcion, which showed good linear relationship in the range of 0.0122–0.7344 μg (r=0.9999), 0.00714–5.7120 μg (r=0.9999), 0.0088–0.7040 μg (r=1.0000), 0.1224–3.6720 μg (r=0.9999) and 0.0148–5.9200 μg (r=0.9997), respectively. There was no significant difference between the calculated value and the measured value by using the relative correction factor (RCF), and the reproducibility of RCF was good. The quality of Rheum palmatum L. from different areas and ages in Gansu Province significantly differed (P<0.05). Two year old Rheum palmatum L. from Pingxiang village of Li county and Rushu village of Tanchang county had the best quality, and that of three years old was better than two years old from Tanchang county. Conclusion The established fingerprints and QAMS method is accurate, feasible, and can be used for the quality comparison of Rheum palmatum L. from different areas and ages in Gansu Province. Li County and Tanchang County areas in Gansu Province can be used as suitable planting areas and development industry. Therefore, it is recommended to select more than two yeas old Rheum palmatum L. from Tanchang County and Li County in Gansu Province for medicinal application.
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A new leaf spot disease was observed on leaves of Rheum palmatum (Chinese rhubarb) in Northwest China (Gansu Province) starting in 2005. A Septoria-like fungus was isolated and completion of Koch's postulates confirmed that the fungus was the casual agent of the leaf spot disease. Morphology and molecular methods were combined to identify the pathogen. The fungus produced conidiomata pycnidia and the conidia were 2~5 septate, 61.2~134.1 µm in length and 3.53~5.3 µm in width, which is much larger than the known Spetoria species that infects Polygonaceae species. Phylogenic analysis of the internal transcribed spacer region confirmed that this Spetoria-like fungus is within the Spetoria genus but distinct from known Spetoria species. Together, these morphological and phylogenetic data support that the R. palmatum infecting Spetoria strain is a newly-described plant pathogenic species.
Sujets)
Chine , Classification , Champignons , Plantes , Polygonaceae , Rheum , Spores fongiquesRésumé
OBJECTIVE:To study the effects of Heishun tablets combined with Rheum palmatum on the pharmacokinetics of hypaconitine in rats. METHODS:Rats were randomly divided into single drug group(Heishun tablets decoction)and drug combi-nation group(Heishun tablets-R. palmatum mixture decoction),with 18 rats in each group. They were given relevant drugs intragas-trically,by 10 g(medicinal materials)/kg of Heishun tablets. 0.3 ml blood samples were collected before(0 h)and 0.083,0.167, 0.333,0.5,0.75,1,1.5,2,3,4,6,8,10 h after medication with 6 rats at each time point,respectively. The blood concentra-tion of hypaconitine was determined by HPLC-MS using palmatine hydrochloride as internal standard. DAS 2.0.1 software was used to calculate pharmacokinetic parameters. RESULTS:The linear range of hypaconitine was 0.102 4-100 ng/ml (r=0.998 7),and the limit of quantification was 0.1 ng/ml. The pharmacokinetic parameters of single drug group vs. drug combination group were as follows as tmax of (0.50 ± 0.086) h vs. (0.75 ± 0.132)h;t1/2 of (9.967 ± 1.123) h vs. (3.708 ± 0.507) h;AUC0-10 h of (26.087 ± 0.672) μg·h/L vs.(6.516 ± 1.135) μg·h/L;cmax of (6.124 ± 2.312) μg/L vs. (1.592 ± 0.051) μg/L. Compared with single drug group,t1/2,AUC0-10 h and cmax of hypaconitine were decreased in drug combination group,with statistical significance (P<0.05). CONCLUSIONS:R. palmatum can inhibit the absorption of hypaconitine in rats,and speed up the elimination of it in rats.