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OBJECTIVE To compare t he diff erences of the fingerprint and in vitro antioxidant activity between decoction pieces of Polygonum cuspidatum by integrated technology of habitat processing and processing （short for IPDP ）and traditional processing decoction pieces （short for TPDP ）. METHODS Ten batches of IPDP and ten batches of TPDP were prepared by integrated technology and traditional technology ，respectively. HPLC method was used to establish and compare the fingerprints of IPDP and TPDP. The scavenging rates of DPPH free radical ，ABTS free radical ，superoxide free radical and hydroxyl free radical and reducing activity of Fe 3+ were detected for IPDP and TPDP. In vitro antioxidant activities were compared between IPDP and TPDP. RESULTS There were 11 common peaks in the fingerprints of IPDP and TPDP ，among which 17 came from IPDP and 13 came from TPDP. The peak heights of peak 6（polydatin）and peak 15（emodin-8-O-β-D-glucoside）in IPDP were significantly higher than those in the TPDP ，and the peak heights of peak 13（resveratrol），peak 17（emodin）and peak 19（physcion）in the TPDP were significantly higher than those in the IPDP. The results of in vitro antioxidant test showed that IPDP and TPDP had a certain scavenging capacity on DPPH free radical ，ABTS free radical ，superoxide free radical and hydroxyl free radicals ，and also had a certain reducing capacity on Fe 3+. CONCLUSIONS The integrated processing technology of P. cuspidatum has a good retention effect on the glycosides in P. cuspidatum ，and the in vitro antioxidant activity of IPDP is stronger than that of TPDP.
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Pulmonary fibrosis(PF)is an irreversible lung disease that is characterized by excessive scar tissue with a poor median survival rate of 2-3 years.The inhibition of transforming growth factor-β receptor type-Ⅰ(TGF-β RI)by an appropriate drug may provide a promising strategy for the treatment of this disease.Polygonum cuspidatum(PC)is a well-known traditional Chinese herbal medicine which has an anti-PF effect.Accordingly,a combination of high resolution mass spectrometry with an in silico strategy was developed as a new method to search for potential chemical ingredients of PC that target the TGF-β RI.Based on this strategy,a total of 24 ingredients were identified.Then,absorption,distribution,meta-bolism,and excretion(ADME)-related properties were subsequently predicted to exclude compounds with potentially undesirable pharmacokinetics behaviour.Molecular docking studies on TGF-β RI were adopted to discover new PF inhibitors.Eventually,a compound that exists in PC known as resveratrol was proven to have excellent biological activity on TGF-β RI,with an ICso of 2.211 μM in vitro.Furthermore,the complex formed through molecular docking was tested via molecular dynamics simulations,which revealed that resveratrol had strong interactions with residues of TGF-β RI.This study revealed that resveratrol has significant potential as a treatment for PF due to its ability to target TGF-β RI.In addition,this research demonstrated the exploration of natural products with excellent biological activities toward specific targets via high resolution mass spectrometry in combination with in silico technology is a promising strategy for the discovery of novel drugs.
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The pathogenesis of nonalcoholic fatty liver disease (NAFLD) remains unclear, and currently no effective drugs have been approved for the treatment of NAFLD. Polygonum cuspidatum is a natural traditional Chinese medicine with a long history of application, and studies have shown that it plays an important role in the treatment of NAFLD. This article summarizes related research findings in the active components of Polygonum cuspidatum applied in the treatment of NAFLD, and it is found that the active components of Polygonum cuspidatum can improve insulin resistance, exert an anti-oxidative stress effect, regulate lipid metabolism, improve endoplasmic reticulum stress, and alleviate inflammatory infiltration by regulating the signaling pathways including Nrf2, AMPK, NF-κB, SIRT1, and PPARα, thereby exerting a preventive and therapeutic effect on NAFLD, so as to provide a basis and ideas for developing drugs for NAFLD and exploring related mechanisms.
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Abstract Gut bacterial β-glucuronidase (GUS) can reactivate xenobiotics that exert enterohepatic circulation- triggered gastrointestinal tract toxicity. GUS inhibitors can alleviate drug-induced enteropathy and improve treatment outcomes. We evaluated the inhibitory effect of Polygonum cuspidatum Siebold & Zucc. and its major constituents against Escherichia coli GUS (EcGUS), and characterized the inhibitory mechanism of each of the components. Trans-resveratrol 4'-O-β-D-glucopyranoside (HZ-1) and (-)-epicatechin gallate (HZ-2) isolated from P. cuspidatum were identified as the key components and potent inhibitors. These two components displayed strong to moderate inhibitory effects on EcGUS, with Ki values of 9.95 and 1.95 μM, respectively. Results from molecular docking indicated that HZ-1 and HZ-2 could interact with the key residues Asp163, Ser360, Ile 363, Glu413, Glu504, and Lys 568 of EcGUS via hydrogen bonding. Our findings demonstrate the inhibitory effect of P. cuspidatum and its two components on EcGUS, which supported the further evaluation and development of P. cuspidatum and its two active components as novel candidates for alleviating drug-induced damage in the mammalian gut.
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OBJECTIVE：To establish the UPLC fingerprint of Polygonum cuspidatum ，and to determine the contents of four active ingredients and to provide reference for the quality evaluation of P. cuspidatum . METHODS ：The determination was performed on Waters BEH C 18 column（100 mm×2.1 mm，1.7 μm）with mobile phase consisted of acetonitrile- 0.2％ formic acid （gradient elution ）at flow rate of 0.4 mL/min. The column temperature was 40 ℃，and detection wavelength was 290 nm. The sample size was 1 μL. The fingerprints were evaluated by similarity calculation，cluster analysis and orthogonal partial least square discriminant analysis （OPLS-DA）. Using polydatin as internal standard ，relative calibration factors of resveratrol ，emodin-8-O- β-D-glucoside and emodin were determined to develop a method of QAMS. The contents of 4 above components in 15 batches of P. cuspidatum were calculated by relative calibration factors. The results of QAMS were compared with those of external standard. RESULTS：UPLC fingerprints of 15 batches of P. cuspidatum were established ，and 12 common peaks were confirmed. Five components were identified ，i.e. polydatin ，resveratrol，emodin-8-O-β-D-glucoside，emodin，emodin methyl ether. The fingerprint similarity of 15 batches of P. cuspidatum was in the range of 0.865-0.976. According to cluster analysis ，15 batches of P. cuspidatum were classified into 4 categories，showing certain regularity of origin. Seven markers were identified by OPLS-DA method. The order of difference significance was peak 7＞emodin-8-O-β-D-glucoside＞resveratrol＞peak 8＞polydatin＞peak 1＞ peak 10. The relative deviation among the contents of resveratrol ，emodin-8-O-β-D-glucoside and emodin in 15 batches of P. cuspidatum determined by QAMS and external standard method was less than 5.0％，indicating that there was no significant difference between the two methods. CONCLUSIONS ：UPLC fingerprint combined with QAMS method is convenient and reliable for the quality evaluation of P. cuspidatum ；the quality of P. cuspidatum produced in Chongqing and Anhui province is better.
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OBJECTIVE: To study the chemical constituents from Polygonum cuspidatum. METHODS: The petroleum ether-ethyl acetate solvents at different ratios of 9:1, 7:1, 5:1, 3:1 and 1:1 were used as gradient eluents for silica gel column chromatography, and samples collected were further isolated and purified by preparative high performance liquid chromatography. Their structures were elucidated by physico-chemical constants and spectroscopic methods. RESULTS: Eight compounds were obtained and identified as 22(Z)-ergosterol-4,6,8,22-tetraene-3-one(1), (22E,24R)-stigma-1,4-diene-3-one(2), (E)-ethyl octadecanoic-16-enoic acid ethyl ester(3), oleanolic acid(4), emodin(5), resveratrol(6), trans-resveratrol glycoside(7) and 6'-gallic acid-4-O-D-resveratrol ester(8).CONCLUSION: Compound 1 is a new natural compound (patent No. ZL 2013 10205435.5), compound 2 and 3 are isolated from this plant for the first time.
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Objective: To identify the transcriptional activity and expression profile of PcMYB1, encoding a new R2R3-MYB transcription factor from Polygonum cuspidatum, and evaluate the biological functions of PcMYB1 in transgenic Arabidopsis thaliana. Methods: The yeast one-hybrid system assay was conducted to test the transcriptional activity of PcMYB1. The tissue-specific expression profiles of PcMYB1 and the gene expression of P. cuspidatum leaves in response to UV-C irradiation were analyzed by RT-PCR analysis. To assess the biological functions of PcMYB1, the gene was expressed in A. thaliana under the control of CaMV 35S promoter. To obtain information about the lignin composition, cross-sections of the basal part of the inflorescence stem of wild-type and transgenic A. thaliana plants were treated with Wiesner staining, and the lignin content was measured by acetyl bromide method. RT-PCR analysis was used to determine expression levels of the genes encoding the enzymes of lignin biosynthesis. Results: After expression of reporter and effector constructed in yeast, β-galactosidase assays showed that the transcriptional activation activity of VP16 domain was reduced markedly when fused to PcMYB1 protein, indicating that PcMYB1 has transcriptional repression activities. Expression pattern analysis showed that PcMYB1 was widely expressed in all tissues examined, but predominantly in leaves. PcMYB1 showed a peak of transcription at 6 h post UV-C treatment. The transgenic lines with reduction in height was 24.07% shorter than the wild-type plants. Wiesner staining of lignin in stem cross-sections revealed the typical intense red stain of secondary cell walls in wild-type plants, but less intense staining was detected in transgenic plants, and lignin accumulation was significantly decreased (about 14.81%) in transgenic plants stems. The expression of genes involved in the lignin biosynthetic pathway, including AtC4H, AtC3H, AtF5H, AtCOMT, and AtCAD, were down-regulated in transgenic lines compared to wild-type plants. Conclusion: Taken together, this study provided the evidence for the biological functions of PcMYB1 as a negative regulator of lignin pathway.
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Objective: To investigate the inhibitory effect and targets of enriched components from Polygonum cuspidatum on HIV-1 in vitro. Methods: Four extracts of P. cuspidatum were screened by HIV-1 multi-target screening system based on the surface plasmon resonance. The highly active components were obtained by NHS-activated HiTrap conjugated with integrase. The anti-HIV activity of the sample was determined with TZM-bl infection assay and PBMCs infection assay. The inhibition of HZ60-IN on integrase 3’ processing was detected by fluorescence resonance energy transfer analysis. High-throughput ELISA was used to determine the effect of enriched active ingredients of P. cuspidatum (HZ60-IN) on integrase chain transfer; Effects of HZ60-IN on reverse transcriptase and protease were detected by kit. Results: HZ60-IN displayed higher affinity with integrase. HZ60-IN demonstrated potent antiviral activity against NL4.3 and 1084i strains in TZM-bl cells with the IC50 of (31.94 ± 8.96) and (38.07 ± 11.25) μg/mL, respectively. HZ60-IN showed significant inhibition on HIV-1 NL4.3 strains in PBMCs from two donors. HZ60-IN acted on the integrase with the IC50 of (6.54 ± 1.69) μg/mL for 3’ processing and (2.56 ± 0.97) μg/mL for strand transfer activity. It showed weak effects on the entry stage of HIV infection, with weak inhibitory activity on reverse transcriptase and no effect on protease activity. Conclusion: HZ60-IN showed significant inhibitive effect on HIV-1 replication and might specifically interfere with integrase activities.
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The aim of this paper was to study the effect and mechanism of alcohol extract from Polygonum cuspidatum(PCE) on acute gouty arthritis in C57 BL/6 mice through NLRP3/ASC/caspase-1 axis. The model mice which injected with ankle joint injection of sodium urate crystals(MSU) were orally administrated with three different concentration of PCE, with colchicine as positive control. HE staining was used for observing the morphological changes of synovial tissue; concentration of IL-1β, IL-6 and TNF-α secreted by synovial tissue of the ankle joint were detected by ELISA; mRNA and protein expression of NLRP3, ASC and caspase-1 in synovial tissue were detected by RT-PCR and Western blot respectively. The results showed that the swelling degree of ankle joint in model mice were significantly elevated; expression of IL-1β, IL-6 and TNF-α were significantly increased; mRNA and protein expression of NLRP3, ASC and caspase-1 also significant increase, compared with normal control group. The swelling degree of ankle joint significantly relief; expression of IL-1β, IL-6 and TNF-α in joint synovium significantly decrease; mRNA and protein expression of NLRP3, ASC and caspase-1 were significantly decrease in PCE treatment group compared with model group. Our research implied that alcohol extract from P. cuspidatum had positive effect on acute gouty arthritis in mice, and the regulation of NLRP3/ASC/caspase-1 axis may be its mechanism.
Subject(s)
Animals , Mice , Ankle Joint , Arthritis, Gouty , Drug Therapy , CARD Signaling Adaptor Proteins , Metabolism , Caspase 1 , Metabolism , Fallopia japonica , Chemistry , Interleukin-1beta , Metabolism , Interleukin-6 , Metabolism , Mice, Inbred C57BL , NLR Family, Pyrin Domain-Containing 3 Protein , Metabolism , Plant Extracts , Pharmacology , Tumor Necrosis Factor-alpha , Metabolism , Uric AcidABSTRACT
BACKGROUND@#To observe the effects of Chinese medicine (CM) Polygonum cuspidatum (PC) on adenosine 5'-monophosphate-activated protein kinase (AMPK), forkhead box O3α (FOXO3α), Toll-like receptor-4 (TLR4), NACHT, LRR and PYD domains-containing protein 3 (NLRP3), and monocyte chemoattractant protein-1 (MCP-1) expression in a rat model of uric acid-induced renal damage and to determine the molecular mechanism.@*METHODS@#A rat model of uric acid-induced renal damage was established, and rats were randomly divided into a model group, a positive drug group, and high-, medium-, and low-dose PC groups (n=12 per group). A normal group (n=6) was used as the control. Rats in the normal and model groups were administered distilled water (10 mL•kg) by intragastric infusion. Rats in the positive drug group and the high-, medium-, and low-dose PC groups were administered allopurinol (23.33 mg•kg), and 7.46, 3.73, or 1.87 g•kg•d PC by intragastric infusion, respectively for 6 to 8 weeks. After the intervention, reverse transcription polymerase chain reaction, Western blot, enzyme linked immunosorbent assay, and immunohistochemistry were used to detect AMPK, FOXO3α, TLR4, NLRP3, and MCP-1 mRNA and protein levels in renal tissue or serum.@*RESULTS@#Compared with the normal group, the mRNA transcription levels of AMPK and FOXO3α in the model group were significantly down-regulated, and protein levels of AMPKα1, pAMPKα1 and FOXO3α were significantly down-regulated at the 6th and 8th weeks (P<0.01 or P<0.05). The mRNA transcription and protein levels of TLR4, NLRP3 and MCP-1 were significantly up-regulated (P<0.01 or P<0.05). Compared with the model group, at the 6th week, the mRNA transcription levels of AMPK in the high- and medium-dose groups, and protein expression levels of AMPKα1, pAMPKα1 and FOXO3α in the high-dose PC group, AMPKα1 and pAMPKα1 in the mediumdose PC group, and pAMPKα1 in the low-dose PC group were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription and protein levels of TLR4 and NLRP3 in the 3 CM groups, and protein expression levels of MCP-1 in the medium- and low-dose PC groups were down-regulated (P<0.01 or P<0.05). At the 8th week, the mRNA transcription levels of AMPK in the high-dose PC group and FOXO3α in the medium-dose PC group, and protein levels of AMPKα1, pAMPKα1 and FOXO3α in the 3 CM groups were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription levels of TLR4 in the medium- and low-dose PC groups, NLRP3 in the high- and low-dose PC groups and MCP-1 in the medium- and low-dose PC groups, and protein expression levels of TLR4, NLRP3 and MCP-1 in the 3 CM groups were down-regulated (P<0.01 or P<0.05).@*CONCLUSION@#PC up-regulated the expression of AMPK and its downstream molecule FOXO3α and inhibited the biological activity of TLR4, NLRP3, and MCP-1, key signal molecules in the immunoinflammatory network pathway, which may be the molecular mechanism of PC to improve hyperuricemia-mediated immunoinflflammatory metabolic renal damage.
Subject(s)
Animals , Male , Rats , AMP-Activated Protein Kinases , Physiology , Chemokine CCL2 , Blood , Disease Models, Animal , Fallopia japonica , Forkhead Box Protein O3 , Physiology , Hyperuricemia , Kidney Diseases , Drug Therapy , Plant Extracts , Pharmacology , Rats, Sprague-Dawley , Signal Transduction , Uric AcidABSTRACT
Objective To prepare solid dispersions (SD) of Polygonum cuspidatum extract (PCE) with the intention of improving the dissolution rate of its active ingredients in vitro and exploring the applicability of hot melt extrusion in SD of traditional Chinese medicine extract. Methods The SD of PCE were prepared via hot melt extrusion based on Eudragit EPO. The cumulative dissolution of resveratrol and emodin in the extract of PCE was used as an evaluation index, and the single factor experiment was optimized. Then physicochemical properties of the SD prepared by optimized process were investigated by differential scanning calorimetry (DSC), powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). Results The optimized process was 120 r/min for screw speed, 160 ℃ for barrel temperature and liquid nitrogen cooling. Under the optimized conditions, the dissolution rate of resveratrol and emodin in the extract was significantly improved, and the drug was present in the amorphous state in the carrier. Conclusion Hot melt extrusion can prepare SD of PCE to improve the dissolution rate of resveratrol and emodin, which provided experimental reference and data support for the study ofpreparation of SD of traditional Chinese medicine by hot melt extrusion.
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Objective To study the fingerprint of Polygonum cuspidatum from various regions by HPLC and provide technical support for the identification.Methods HPLC conditions :the chromatographic column was Agilent ZORBAX SB-C18 (4.6 mm × 250 mm,5 μm);the mobile phase was 0.1% formic acid in water and acetonitrile (A-B);gradient elution ;the de-tection wavelength was 306 nm ;the flow rate was 1 ml/min;the column temperature was 30 ℃ ;injection volume was 20 μl. Results The HPLC gradient conditions could effectively separate polydatin,resveratrol,emodin and emodin ether,etc.Con-clusion This method is stable,effective and durable,which is instructive to the herbs data identification of Polygonum cuspi-datum.
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Anti-Helicobacter pylori activity guided fractionation led to the isolation of five anthraquinones, two stilbenes and one naphthoquinone from the EtOAc fraction of Polygonum cuspidatum, using silica gel column chromatography, Sephadex-LH20, MPLC and recrystallization. The chemical structures were identified to be physcion (1), emodin (2), anthraglycoside B (3), trans-resveratrol (4), anthraglycoside A (5), polydatin (6), 2-methoxy-6-acetyl-7-methyljuglone (7) and citreorosein (8) by UV, ¹H-NMR, ¹³C-NMR and mass spectrometry. Anti-Helicobacter pylori activity including MIC values of each compound was evaluated. All of the isolates exhibited anti-H. pylori activity of which MIC values were lower than that of a positive control, quercetin. Compounds 2 and 7 showed potent growth inhibitory activity. Especially, a naphthoquinone, compound 7 displayed most potent antibacterial activity with MIC₅₀ value of 0.30 µM and MIC₉₀ value of 0.39 µM. Although anti-H. pylori activity of this plant was previously reported, this is the first report on that of compounds isolated from this species. From these findings, P. cuspidatum roots or its isolates may be useful for H. pylori infection and further study is needed to elucidate mechanism of action.
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Anthraquinones , Chromatography , Emodin , Fallopia japonica , Mass Spectrometry , Plants , Polygonum , Quercetin , Silica Gel , StilbenesABSTRACT
The present study investigated the chemical composition of ethylacetate extracts from an endophytic actinomycete Streptomyces sp. A0916 and its host Polygonum cuspidatum. A comparative analysis of the antimicrobial and antioxidant properties of the extracts was also conducted. 32 compounds of P. cuspidatum and 23 compounds of Streptomyces sp. A0916 were isolated and identified by GC/MS. Antimicrobial activities of the extracts were evaluated using eight microbial strains (3 Gram-positive bacteria, 3 Gram-negative bacteria, and 2 fungi). The Streptomyces sp. A0916 extracts showed a wide range of antimicrobial activities and presented greater antimicrobial effectiveness than the P. cuspidatum extracts. The minimum inhibitory concentration (MIC) of Streptomyces sp. A0916 extracts against the ampicillin-resistant strain Enterococcus faecium SIIA843 was 32 μg·mL(-1). Furthermore, the extracts had greater antimicrobial effect against Gram-positive bacteria than Gram-negative bacteria. Finally, the antioxidant activity of the Streptomyces sp. A0916 extracts was equal to that of the P. cuspidatum extracts. In conclusion, our results suggest that the endophytic actinomycetes of the medicinal plants are an important source of bioactive substances.
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Anti-Infective Agents , Chemistry , Pharmacology , Antioxidants , Chemistry , Pharmacology , Bacteria , Fallopia japonica , Chemistry , Microbiology , Fungi , Plant Extracts , Chemistry , Pharmacology , Streptomyces , Chemistry , Classification , GeneticsABSTRACT
Objective To evaluate the effect of polydatin on neuropathic pain in rats.Methods Forty male Sprague-Dawley rats,aged 6-8 weeks,weighing 200-230 g,were randomly divided into 5 groups (n =8 each) using a random number table:sham operation group (group S),neuropathic pain group (group NP),polydatin 5 mg/kg group (group P1),polydatin 10 mg/kg group (group P2),and polydatin 20 mg/kg group (group P3).Neuropathic pain was induced by chronic constriction injury in NP and P1-3 groups.In group S,the sciatic nerve was only exposed but not ligated.In S and NP groups,normal saline 0.1 ml was injected intraperitoneally immediately after operation and at 1,3,5 and 7 days after operation (T1-4).In P1-3 groups,polydatin 5,10 and 20 mg/kg (in normal saline 0.1 ml) were injected intraperitoneally immediately after operation and at T1-4.At 1 day before operation (T0) and T1-4,the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.After measurement of pain threshold at T4,the rats were sacrificed,and L4-6 segments of the spinal cords were removed for determination of the expression of high-mobility group box 1 (HMGB1),Toll-like receptor 4 (TLR4),interleukin-1beta (IL-1β),tumor necrosis factor-alpha (TNF-α) and monocyte chemotactic protein-1 (MCP-1) by Western blot.Results Compared with group S,the MWT was significantly decreased,and the TWL was significantly shortened at T1-4 in group NP,the MWT was significantly decreased at T1-4,and the TWL was significantly shortened at T2-4 in group P1,the MWT was significantly decreased at T1-4,and the TWL was significantly shortened at T3.4 in group P2,the MWT was significantly decreased at T1-4 in group P3,and the expression of HMGB1,TLR4,IL-1β,TNF-α and MCP-1 was significantly up-regulated in NP,P1 and P2 groups (P＜0.05).Compared with group NP,the MWT was significantly increased at Tt-4,and the TWL was significantly prolonged at T1,2 in group P2,the MWT was significantly increased,and the TWL was significantly prolonged at T1-4 in group P3,the expression of HMGB1,TLR4,IL-1β,TNF-α and MCP-1 was significantly down-regulated in P2 and P3 groups (P＜0.05),and no significant change was found in the parameters mentioned above in group P1 (P＞0.05).Compared with group P1,the MWT was significantly increased at T4 in group P2,and the MWT was significantly increased at T14,the TWL was significantly prolonged at T3,4,and the expression of HMGB1,TLR4,IL-1β,TNF-α and MCP-1 was significantly down-regulated in group P3 (P＜0.05).Compared with group P2,the MWT was significantly increased at T3,4,and the expression of TLR4,IL-1β,TNF-α and MCP-1 was significantly down-regulated in group P3 (P＜0.05).Conclusion Polydatin can alleviate neuropathic pain through inhibiting inflammatory responses in the spinal cord of rats.
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Objective:To establish the HPLC fingerprint for Polygonum cuspidatum dispensing granules. Methods:The HPLC fin-gerprint of 12 batches of Polygonum cuspidatum from different manufacturers were determined. The analysis was performed on a Waters SunFire C18 column(250 mm × 4. 6 mm,5μm)with acetonitrile as the mobile phase A and 0. 05% phosphoric acid solution as the mo-bile phase B with gradient elution. The flow rate was 1. 0 ml·min-1 ,the detection wavelength was 230 nm,the column temperature was 30℃,and the injection volume was 10μl. Results:The results were calculated according to“similarity evaluation system for tradi-tional Chinese medicine chromatographic fingerprint”nominated by CFDA combined with the analysis of the HPLC fingerprints. Totally 14 common peaks with similarity above 0. 98 were found in the HPLC fingerprint of Polygonum cuspidatum,including the peak respec-tively for polydatin and emodin. Conclusion:The method can provide more information for the quality control of Polygonum cuspidatum dispensing granules.
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Objective: To construct the RNAi expression vector of Polygonum cuspidatum chalcone synthase (PcCHS1) gene, and to obtain the transgenic plants in which PcCHS1 expression was down-regulated. Methods: According to known sequence (EF090604) of PcCHS1 gene in GenBank, right primers were designed and the conserved sequence was cloned. The conserved fragment (574 bp) targeting at PcCHS1 gene was inserted into the expression vector pYLRNAi in both forward and reverse directions, and RNA interference (RNAi) expression vector pYLRNAi-PcCHS1 was constructed. Using the method of Agrobacterium-mediated transformation, the expression vector was used to transform the shoot tips of P. cuspidatum, and transgenic plants were obtained. The expression of PcCHS1 was confirmed by Northern blotting and the accumulation of polydatin was detected by HPLC. Results: RNAi expression vector of PcCHS1gene was constructed successfully, and five transgenic plants were obtained. Northern blotting analyses indicated that the expression levels of PcCHS1 were significantly down-regulated in the transgenic plants. Polydatin concentration in the transgenic plants was up to 3.8 times higher than that in non-transformed control plants. Conclusion: Transgenic P. cuspidatum plants with down-regulated expression of PcCHS1 gene were obtained successfully. The content of polydatin in the transgenic P. cuspidatum was significantly increased by RNAi against PcCHS1. This work might establish an experimental basis for the effective application of PcCHS1 in improving polydatin accumulation in P. cuspidatum.
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Objective: To establish the chromatography fingerprint of Polygoni Cuspidati Rhizoma et Radix with hyphenated technique of HPLC-DAD-ELSD and to evaluate Polygoni Cuspidati Rhizoma et Radix from 10 different origins. Methods: Luna C18 (2) column (250 mm × 4.6 mm, 5 μm) was used. Mobile phase was acetonitrile-H2O; Flow speed was 1.0 mL/min; Temperature of column was set at 35℃; Detective wavelength was at 254 nm; Injection volume was 10 μL. The temperature of drift tube was 109℃ and the flow speed was 3.0 L/min. Results: The chromatography fingerprint of Polygoni Cuspidati Rhizoma et Radix from 10 different origins was established. In the chromatography fingerprint with HPLC-DAD of Polygoni Cuspidati Rhizoma et Radix, 19 common peaks were demarcated and the similarities of Polygoni Cuspidati Rhizoma et Radix were between 0.938-0.993. In the chromatography fingerprint with HPLC-ELSD of Polygoni Cuspidati Rhizoma et Radix, 14 common peaks were demarcated and the similarities of Polygoni Cuspidati Rhizoma et Radix were between 0.905-0.999. Polydatin, resveratrol, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside, emodin, and physcion were identified. Conclusion: The method is accurate and stable, which can be used as the evidence for the quality evaluation of Polygoni Cuspidati Rhizoma et Radix.
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Objective: To investigate the chemical constituents in the flowers of Polygonum cuspidatum. Methods: The components were separated by means of solvent extraction, repeated chromatography with silica and Sephadex LH-20 column. The structures were determined by spectral analysis and physicochemical properties. Results: Sixteen compounds were isolated from the ethyl ether extract and methanol extract from the flowers of P. cuspidatum and identified as β-sitosterol (1), aloe-emodin (2), physcion (3), emodin (4), daucosterol (5), chrysophanol (6), luteolin (7), kaempferol (8), anthraglycoside B (9), rhein (10), apigenin (11), hesperetin (12), 4-hydroxyacetophenone (13), rutin (14), sucrose (15), and genistein (16). Conclusion: Compounds 2, 8, 12, 13, 15, and 16 are obtained from the flowers of P. cuspidatum for the first time.
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Objective:To compare the content of polydatin and emodin between polygonum cuspidatum pieces and dispensing gran-ules to provide theoretical basis for the improvement of preparation process of the granules. Methods:An HPLC method with linear gra-dient elution was used to determine the content of polydatin and emodin in the pieces and dispensing granules, and the difference in the two contents was compared. Results: The content of polydatin and emodin in polygonum cuspidatum pieces met the requirement de-scribed in China pharmacopoeia, which was lower than that in the dispensing granules, and the contents of the two components in the commercial dispensing granules were not equivalent to the labeled amount, which was 1/15 and 1/27 of that in the pieces. Conclu-sion:The transport rate of the traditional preparation process of dispensing granules is with poor efficiency, which should be improved.