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The traditional Chinese medicine Aralia elata (Miq.) Seem., also known as Aralia mandshurica, has the effect of "tonifying Qi and calming the mind, strengthening the essence and tonifying the kidneys, and dispelling wind and invigorating blood circulation". It is used in the treatment of neurasthenia, Yang deficiency and Qi deficiency, kidney Qi deficiency, spleen Yang deficiency, water-dampness stagnation, thirst, and bruises. Aralia elata saponins are the main components for the pharmacological effects. From the perspective of modern pharmacological science, Aralia elata has a wide range of effects, including anti-myocardial ischaemia and alleviation of secondary myocardium ischemic reperfusion injury by regulating ionic homeostasis, anti-tumor activity by inhibiting proliferation, promoting apoptosis and enhancing immunity, hypoglycemia and lipid lowering effects by regulating glucose and lipid metabolism, and hepato-protective, neuroprotective, anti-inflammatory/analgesic effects. The studies on pharmacological mechanisms of Aralia elata will be conducive to its development and application in the future. This article reviews the research progress of Aralia elata domestically and internationally in the last two decades and proposes new directions for further research.
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Aralia , Yang Deficiency , Apoptosis , Saponins/pharmacology , Myocardial IschemiaABSTRACT
OBJECTIVE@#Pseudostellaria heterophylla has been paid more attention in recent years, mainly as a medicine food homology plant. The content determination of P. heterophylla is not specified in the Chinese Pharmacopoeia (version 2020). The environmental conditions in different production areas could exert an influence on the quality of P. heterophylla. The purpose of this study is to discriminate P. heterophylla collected from different geographical origins of China.@*METHODS@#In this study, the content of polysaccharide in 28 batches of P. heterophylla was determined using phenol-sulfuric acid. HPLC fingerprints were established under optimised HPLC-PDA methods. Subsequently, the similarity analysis (SA) and the quantification of heterophyllin B were analyzed. The metabolites of P. heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and orthogonal PLS-DA (OPLS-DA) were performed based on all peak areas.@*RESULTS@#The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas, which varied significant from different origins. While the content of heterophyllin B in Anhui and Jiangsu was high. The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990, and the characteristic map can be used to identify and evaluate the quality of P. heterophylla. The samples from Fujian, Guizhou, Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA, PLS-DA, HCA, OPLS-DA, indicating that their metabolic compositions were significantly different. Ultimately, a total of 15 metabolites which were filtrated by a VIP-value > 1 and a P-value < 0.05 associated with the separation of different origins were identified.@*CONCLUSION@#HPLC fingerprint was established to evaluate the quality and authenticity of P. heterophylla. The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions. A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P. heterophylla from different origins in China for the first time. Overall, this study provides insights to metabolomics of P. heterophylla and supplies important reference values for the development of functional foods.
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Squalene is a key metabolic intermediate for sterols and various other triterpenoids. Its biosynthesis is catalyzed by squalene synthase (SQS), which converts two molecules of farnesyl pyrophosphate to squalene. The biosynthetic pathway of Fritillaria thunbergii Miq isosteroid alkaloids is similar to that of triterpenoids. In this study, a full-length cDNA of squalene synthase from Fritillaria thunbergii Mig (FtSQS) was cloned using rapid amplification from cDNA ends (RACE) technology. GenBank accession number was KF551097. 2. Bioinformatics methods were used to characterize the FtSQS in detail, including the detection of conserved regions, sequence homology analysis, secondary and tertiary structure prediction, and phylogenetic tree analysis. The results showed that its open reading frame (ORF) was 1 230 bp and encoded 409 amino acids. Protein-Blast alignment found that amino acid homology with SQS of Indian pine, Truncate alfalfa, Purple shirt, Potato, Bupleurum, Golden iron lock and Arabidopsis reached 73. 84%, 73. 23%, 72. 24%, 70. 66%, 70. 66%, 69. 44%, 68. 14%. Promoter analysis indicated that the 5' upstream region of FtSQS possessed various potential elements associated with physiological and environmental factors. To obtain a soluble recombinant protein, 24 hydrophobic amino acids were deleted from the carboxyl terminus, and the C-terminal truncated mutant FtSQS (FtSQSATM) was expressed in E. coli BL21 (DE3). SDS-PAGE analysis suggested that approximately 66 kD recombinant protein was checked. The in vitro enzymatic reaction proved that FtSQS could catalyze farnesyl pyrophosphate to generate squalene. Expression level of FtSQS mRNA in leaves was the highest, followed by stem and root, but in bulb was much lower than that in other tissues. It suggests that leaves are active organ for biosynthesis of peimine. The identification and function of FtSQS provides an important basis for the study of secondary metabolites of Fritillaria thunbergii Miq.
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Objective: To establish a rapid and accurate analytical method for the identification of complex system of traditional Chinese medicine, and to systematically clarify the chemical composition of sesquiterpenes in Alpinia oxyphylla. Method: On the basis of optimizing the extraction process of sesquiterpenes, the accurate molecular weight and secondary fragment ions information of unknown compounds were captured by ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Compared with the relative retention time and mass spectrometry data of the reference substance, combined with relevant references and databases, the sesquiterpene unknown compounds in the fruits of A. oxyphylla were accurately and rapidly characterized. Results: A total of 24 sesquiterpenes were identified and classified into four categories according to their skeleton structure, including nine eudesmane-type, six cadinane-type, eight eremophilane-type, and one oplopanone-type. Conclusion: In this study, the established analytical method was used to realize the rapid and accurate identification of sesquiterpenes in the fruits of A. oxyphylla, which provided a theoretical basis for the research on the pharmacodynamic substance basis and quality control of the fruits of A. oxyphylla.
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Objective: To study the secondary metabolites of Penicillium oxalicum, an endophytic fungus isolated from the medicinal plant Pseudostellariaheterophylla. Methods: The secondary metabolites were isolated by silica gel, Sephadex LH-20 column chromatographies, and prep-TLC methods. Their structures were elucidated by using various spectroscopic techniques including HRESIMS and NMR spectra. Results: A benzofuran compound 5,7-dihydroxy-2-methylbenzofuran-3-carboxylic acid (1), five isocoumarins including (3R,4R)-(-)-4-hydroxymellein (2), O-methylmellein (3), acremonone G (4), decarboxycitrinone (5) and decarboxyhydroxycitrinone (6), four bisabolane-type sesquiterpenoids including (7S,11S)-(+)-12-acetoxysydonic acid (7), (S)-(+)-11-dehydrosydonic acid (8), sydonic acid (9) and 1-hydroxy-boivinianin A (10), as well as two meroterpenoid-type alkaloids decaturin D (11) and decaturinC (12) were isolated. Conclusion: Compound 1 is a new benzofuran compound and named as oxafuranone A, while compounds 2-6 and 10 are characterized from P. oxalicum for the first time.
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Objective: To establish an HPLC method for the simultaneous determination of syringin, chlorogenic acid, caffeic acid, p-coumaric acid, ferulic acid, ethylsyringin, ethylconiferin, (+)-fraxinol, (±)-rosin, (±)-syringin, taiwanin C, savinin, helioxanthin, and (-)-sesamin in 16 species and 33 batches of Acanthopanax plants. Methods: Separation was carried out on Promosil C18 column (250 mm × 4.6 mm, 5 μm), the mobile phase was eluted with acetonitrile-0.1% phosphoric acid. The flow rate was 1 mL/min, and the column temperature was 30 ℃, the detection wavelength was 280 nm. Results: Under the optimized chromatographic conditions, the linear relationship of the 14 constituents was good in the range of mass concentration, r was more than 0.999 1, with good precision, stability, repeatability and recovery. Conclusion: HPLC method established in this study is effective, accurate, and reproducible and can be used for the simultaneous determination of phenylpropanoids in Acanthopanax plants, which can provide reference for the further study of Acanthopanax plants.
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Objective: To study the chemical constituents in the fruits of Actinidia arguta. Methods: The compounds were isolated by column chromatography on silica gel, ODS, and Sephadex LH-20, preparative TLC, and semi-preparative HPLC. The structures were established by the analyses of the spectroscopic data. Results: Fifteen compounds were obtained from the n-BuOH fraction of the 75% ethanol extract of the fruits of A. arguta and identified as (2R,6R,9R)-trihydroxy-megastigmane-4,7E-dien-3-one-9-O-β-D- glucopyranoside (1), (6S,9R)-roseoside (2), quercetin-3-O-b-D-galactopyranside (3), astragalin (4), vanillic acid-4-O-β-D- glucopyranoside (5), 1-O-feruloyl-β-D-glucopyranoside (6), ferulic acid-4-O-β-D-glucopyranoside (7), rhodioloside (8), 3-hydroxy-1-(4-O-β-D-glucopyranosyl-3- methoxyphenyl) propan-1-one (9), 5-O-caffeoyl quinic acid methyl ester (10), 5-O-caffeoyl quinic acid butyl ester (11), 5-O-feruloyl quinic acid methyl ester (12), 5-O-coumaroyl quinic acid methyl ester (13), caffeic acid (14), and protocatechuic acid (15). Conclusion: Compound 1 is a new norsesquiterpene glycoside with the megastigmane scaffold, named actinargutaside A. Compounds 2, 5, and 7-13 are isolated from the Actinidia genus for the first time and compound 6 is firstly isolated from A. arguta.
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Objective: To study the chemical constituents of Alpinia oxyphylla. Methods: The ethyl acetate fraction of 95% ethanol extract from A. oxphylla was isolated and purified by silica, MCI, Sephadex LH-20 and semi-preparative HPLC, then the structures of obtained compounds were identified by physicochemical properties and spectral data. Results: Sixteen compounds were isolated and their chemical structures were identified as phthalic acid-bis (2’-ethyl heptanyl) easter (1), (E)-1-(4’-hydroxy-3’-methoxyphenyl)-7- (4″-hydroxy-phenyl)-hept-4-en-3-one (2), 5-hydroxy-7-(4″-hydroxy-3″-methoxyphenyl)-1-phenyl-3-heptanone (3), 1β,4β,7β- trihydroxyeudesmane (4), bullatantriol (5), 1,5-epoxy-3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxy- phenyl) heptanes (6), 1-tetratriacontanol (7), dihydrogingerenone B (8), tectochrysin (9), chrysin (10), oxyphyllenone B (11), (1R,4R,10R)-1β,4α-dihydroxy-11,12,13-trinor-5,6-eudesmen-7-one (12), daucosterol (13), 1-(4’-hydroxyphenyl)-7-(3″-methoxy- 4″-hydroxyphenyl)-4-ene-3-heptanone (14), yakuchinone A (15) and 5-dehydroxy-hexahydro-demethoxycurcumin B (16). Conclusion: A total of 16 compounds were isolated and identified. The compounds 1-2, 4-8 are isolated from the genus for the first time, and compounds 3 were isolated from the plant for the first time.
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Na região serrana do estado do Rio de Janeiro são comuns a prática da truticultura e o consumo dessa espécie de peixe. A aceitação do pescado está estritamente relacionada às características de frescor do mesmo, que pode sofrer alterações devido a processos físico-químicos, microbiológicos e sensoriais de deterioração. Objetivou-se com esse trabalho avaliar o frescor de trutas arco-íris (Oncorhynchus mykiss) e, com isso, desenvolver um padrão específico através da adaptação do Método do Índice de Qualidade (MIQ) para essa espécie, ao longo de 11 dias de armazenamento sob refrigeração. A análise consistiu na avaliação de características determinantes para o frescor das trutas e o registro dos parâmetros foi realizado por meio de um sistema de pontuação de demérito, tendo em vista que quanto menor a pontuação, mais fresco o peixe. Foi concluído que, aos 11 dias, a truta inteira não apresentava aspectos de repugnância ou rejeição de acordo com os parâmetros preconizados pelo RIISPOA.
Subject(s)
Animals , Cooled Foods , Meat/analysis , Food Quality , Oncorhynchus mykissABSTRACT
Objective: To investigate and analyze antioxidant activities in vitro and the active ingredients of the methanol extract of Actinidia arguta. Methods: In this study, antioxidant activities of extract of A. arguta were carried out by using DPPH and ABTS radical scavenging assays in vitro. Qualitative analysis of major active components was performed by HPLC-DAD-ESI-MS/MS. Results: Extract of A. arguta had good scavenging effect on DPPH and ABTS free radical in vitro, and the EC50 values of scavenging effect of extract of A. arguta on DPPH and ABTS free radical were (26.275 ± 1.464) and (29.826 ± 1.309) mg/mL, respectively. On the basis of UV and mass spectral analysis, a total of 19 chemical compositions were preliminarily identified. Conclusion: extract of A. arguta has good antioxidant activity in vitro, and polyhydroxyl and unsaturated double bonds are the main active constituents.
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Objective: To clone the enzyme genes related with methyl eugenol synthesis and characterize the corresponding sequence information based on the transcriptome sequencing of Asarum heterotropoides. Methods: RT-PCR was performed to obtain the full length cDNA of the phenylalanine lyase (PAL) gene, cinnamic acid-4-hydroxylase (C4H) gene, 4-hydroxycinnamoyl-CoA ligase (4CL), and cinnamyl alcohol dehydrogenase (CAD) genes by using young leaves as materials. The acquired enzyme genes were analyzed by bioinformatics. Results: The ORF lengths of AhPAL, AhC4H, Ah4CL, and AhCAD were 2 157, 1 278, 1 623 and 1 071 bp, which respectively encoded 718, 425, 540, and 356 amino acids. Four proteins had respective conserved domains. The amino acid sequences of AhPAL, Ah4CL, and AhCAD were similar to those of other reported species except for AhC4H. Conclusion: Four enzyme genes related with methyl-eugenol synthesis in A. heterotropoides were separated and analyzed using bioinformatics method. These results would lay the important foundation for functional analysis of corresponding genes and for elucidating the regulation mechanism of methyl-eugenol biosynthesis.
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Objective To analyze the material basis and molecular mechanisms of Danggui Beimu Kushen (DBK) Pills in treating prostatic diseases based on the method of integrated pharmacology. Method The platform of Integrative Pharmacology of Traditional Chinese Medicine (TCM-IP, www.tcmip.cn) was utilized to predict the main active ingredients and functional targets of DBK Pills in treating prostatic disease, key targets were screened for enrichment analysis of pathways, and the network of “herb-core component-key target-main pathway” was constructed, and the possible mechanisms of DBK Pills in treating prostatic diseases were explored. Results A total of 532 candidate key targets for the treatment of prostatic diseases by DBK Pills were predicted, and 1 840 terms of gene function and 194 signal pathways were analyzed by gene ontology (GO) and KEGG, respectively. The network analysis of “herb-core component-key target-main pathway” showed that 65 core components were predicted, including 29 ingredients from Angelica sinensis, 11 from Fritillaria thunbergii and 26 from Sophora flavescens. Those predicted components acted on the key targets of prostatic diseases, such as transcription factor binding, negative regulation of apoptosis, et al, through the estrogen, apoptosis, chemokines and other signal pathways, and thus played a role in the regulation of cell cycle, apoptosis and proliferation imbalance, which might be the molecular mechanisms of DBK Pills for the treatment of prostatic disease. Conclusion DBK Pills regulate the development of BPH, prostate cancer and other diseases through multiple pathways with multi-component interacting with multiple targets.
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Objective: Based on structural modification of monomeric compound calenduloside E from Aralia elata, to evaluate anti-inflammatory activity of the analogues. Methods: Applying oleanolic acid as starting material, the target compounds were prepared by seven steps reactions and evaluated for anti-inflammatory effects by RAW264. 7 cells in vitro. Results: Ten analogues G1-G5 and H1-H5 were synthesized. The structures of the target compounds were identified by spectrum. Pharmacological results showed that all of the compounds had different levels potency of anti-inflammatory effects in cells. In particular, compounds G1-G4 and H1-H3 showed significant anti-inflammatory activity comparing wiht lead compounds. Conclusion: The new compounds G1-G5 and H1-H5 which showed potential of anti-inflammatory biological activity, had not been reported in any literatures and deserved further research.
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Objective: To clone the full-length cDNA sequences of psoralen synthase (PS) genes from Glehnia littoralis so as to perform the bioinformatic and expression pattern analysis. Methods: Based on our previous transcriptome sequencing data of G. littoralis, the gene sequences GlPS1 and GlPS2 with high expression level were screened. The 3’cDNA ends of GlPS1 and GlPS2 genes were cloned by the RACE (rapid amplification of cDNA ends) method and the full-length cDNA of genes were assembled by using DNAMAN software. And then encoded GlPS proteins were analyzed by the bioinformatics tools. The issue specific expression of GlPS1 and GlPS2 genes were detected using qPCR. Results: The full-length cDNA of GlPS1 gene was 1 885 bp, which encoding a protein of 495 amino acids with a predicted molecular weight of 55 740.7 and isoelectric point of 8.28; The full-length cDNA of GlPS2 gene was 1 971 bp, which encoding a protein of 502 amino acids with a predicted molecular weight of 56 363.9 and isoelectric point of 6.62. GlPS1 and GlPS2 proteins belong to the cytochrome P450 superfamily, which share one transmembrane zone acting as hydrophilic protein. Phylogenetic analysis showed GlPS1 and GlPS2 were genetically closely related to the PS of Pastinaca sativa, Apium graveolens, Ammi majus. Higher expression of GlPS1 gene was observed in roots than leaves. However, GlPS2 gene was expressed at a relatively higher level in flowers than in roots. Conclusion: The full-length cDNA of GlPS1 and GlPS2 genes were obtained and the expression patterns were explored in G. littoralis for the first time, which provided a foundation for further studies on gene function and genetic regulatory mechanism of GlPS.
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Objective: To evaluate the antioxidant activity of extracts and fractions from Stachys sieboldii Miq., and to examine its effect on the cellular reactive oxygen species (ROS) and glutathione (GSH) production and genomic DNA oxidation in HT-1080 cells. Methods: The ROS generation induced by H
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Objective: To evaluate the antioxidant activity of extracts and fractions from Stachys sieboldii Miq., and to examine its effect on the cellular reactive oxygen species (ROS) and glutathione (GSH) production and genomic DNA oxidation in HT-1080 cells.Methods: The ROS generation induced by H2O2 was measured by the dichlorofluorescein-diacetate assay. GSH levels were measured using a fluorescent method with mBBr. Genomic DNA oxidative damage was measured with levels of oxidative DNA induced by the reaction of ferritin with H2O2.Results: Then-hexane, 85％ aqueous methanol andn-butanol fractions (0.05 mg/mL concentrations) inhibited H2O2-induced ROS generation by 63％, 35％ and 45％, respectively. GSH levels were significantly increased in both acetone+methylene chloride and methanol extracts (P<0.05). Supplementation of cells withn-hexane significantly increased GSH levels at concentrations of 0.05 mg/mL (P<0.05). Both the acetone+methylene chloride and methanol extracts, as well as all fractions significantly inhibited oxidative DNA damage (P<0.05). Conclusions: These results indicate that cellular oxidation was inhibited by then-hexane fraction and this fraction may contain valuable active compounds.
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Objective: To obtain the transcriptome sequence database and differentially expressed genes of Asarum sieboldii, and to identify the genes related to the biosynthesis of methyleugenol, the main chemical compound in this species. Methods: Roots and leaves of the plants were chosen as experiment materials. The transcriptome sequence database was constructed by applying an Illumina Hiseq 4000 Sequencing Platform. Unigenes were assembled by BLAST similarity searches and annotated with GO and KEGG orthologs identifiers. Moreover, differentially expressed genes were analyzed. Results: 12.25 Gb database was obtained, among which 129 003 unigenes were annotated to be involved in 52 GO-terms and 363 metabolic pathways. After analysis, 439 differentially expressed genes were observed, the up-regulated genes account for 38.3% and the down-regulated genes account for 61.7%. In addition, 136 unigenes involved in phenylpropanoid biosynthesis in A. sieboldii, and 44 unigenes that were associated with biosynthesis of methyleugenol were identified. Conclusion: Unigenes explored in this study will significantly contribute to genome-wide research and analysis of this species.
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Objective: To study the influence of sulfur-fumigation on pharmacokinetic of peimine and peiminine of Fritillaria thunbergii in rat plasma by LC-MS/MS. Methods: After random grouping, 18 SD rats were given the solution of fresh-cut and sulfur-fumigated F. thunbergii by ig administration. The blood drug concentration of peimine and peiminine in rat plasma was determined by HPLC-MS/MS, and the pharmacokinetic parameters were calculated with 3P97 software. Results: The pharmacokinetic parameters of peimine and peiminine of sulfur-fumigated F. thunbergii in rat plasma were (66.40 ± 4.65), (146.72 ± 10.88) ng/mL for Cmax, and (181.79 ± 7.85), (457.38 ± 58.81) ng∙h/mL for AUC, respectively. Those of fresh-cut sample in rat plasma were (186.37 ± 18.8), (227.65 ± 7.01) ng/mL for Cmax, and (197.70 ± 18.69), (566.16 ± 41.55) ng∙h/mL for AUC, respectively. The pharmacokinetic parameters of perimine and peiminine of sulfur-fumigated sample in rat plasma were less than those of fresh-cut sample. Conclusion: The results showed that sulfur-fumigation decreased the bioavailability of peimine and peiminine. This study could provide a basis for further clarifying the influence of sulfur-fumigation on efficacy material base of F.thunbergii.
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Objective: To clone the full-length cDNA encoding squalene epoxidase 1 (SQE1), a key enzyme of triterpenes biosynthesis, from Pseudostellaria heterophylla and to perform functional analysis. Methods: With the total RNA as template, the full-length cDNA of SQE1 in P. heterophylla was cloned via RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The bioinformatics of the cloned SQE1 gene was performed. The target gene was transfered into tobacco by Agrobacterium-mediated transformation. Results: The full-length cDNA (2 038 bp) of SQE1 gene was obtained with an open reading frame of 1 554 bp, encoding 517 amino acid polypeptides, which had higher homology with the known SQEs in other medicinal species. The calculated relative molecular mass was 5.67 × 104, the isoelectric point was 8.8. The deduced protein sequence exhibited FAD-binding domains and four transmembrane regions. The content of total triterpenes was increased in transgenic tobacco plants. Conclusion: This is the first report that the full-length cDNA encoding SQE1 from P. heterophylla is cloned. The ectopic expression of SQE1 could promote to increase the content of total triterpenes in transgenic plant. This work provides a foundation for exploring the biosynthetic pathway of triterpenes in P.heterophylla and their applications in bioengineering.
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Peucedanum praeruptorum is a traditional Chinese medicine, which has been recorded in Chinese Pharmacopoeia. Phytochemical studies have shown that the active ingredients of P. praeruptorum can treat anemopyretic cold and cough with abundance of phlegm. Furthermore, the herb is well-known for the treatment of respiratory diseases as well as pulmonary hypertension, and shows great medical values involved in cardiovascular diseases, especially. This paper shows several research achievements involved in plant resources and its adulterants and substitutes identification based on pertinent literatures. DNA barcoding technology shows strong discriminability for medicinal plant and medicinal materials identification of Peucedani Radix. In this paper, recent research advances for identificating Peucedani Radix adulterants and substitutes were elaborated based on modern molecular biotechnology and traditional identification method, merits and drawbacks and practicability of each method are summarized at the same time. The main purpose is to provide new research ideas and solutions for rapid, reliable, and accurate identification of Peucedani Radix.