Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration ...Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Iα-loop-helix Iβmotif which corresponds to helix I of other P 450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,theγ-domain.The structure reveals a homodimer in which theγ-domain of one molecule interacts with theβ-domain of another.The plane of heme group makes an angle of approximately 40°with the helix Iα-loop-helix Iβmotif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recognition.Phe-301,Ser-303 and Gly-305 from the helix Iα-loop-helix Iβmotif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.展开更多
Soybean seed isoflavones are a type of secondary metabolites that can provide health and nutrition benefits for humans. In our previous study, a stable quantitative trait locus(QTL) qIF05-1 controlling the seed isofla...Soybean seed isoflavones are a type of secondary metabolites that can provide health and nutrition benefits for humans. In our previous study, a stable quantitative trait locus(QTL) qIF05-1 controlling the seed isoflavone content in soybean was detected on chromosome(Chr.) 05 in a recombinant inbred line(RIL) population from a cross of Huachun 2×Wayao. In this study, the parental lines were re-sequenced using the Illumina Solexa System with deep coverage. A total of 63,099 polymorphic long insertions and deletions(InDels)(≥15 bp)were identified between the parents Huachun 2 and Wayao. The InDels were unevenly distributed on 20chromosomes of soybean, varying from 1,826 in Chr. 12 to 4,544 in Chr. 18. A total of 10,002 long InDels(15.85% of total) were located in genic regions, including 1,139 large-effect long InDels which resulted in truncated or elongated protein sequences. In the qIF05-1 region, 68 long InDels were detected between the two parents. Using a progeny recombination experiment and genotype analysis, the qIF05-1 locus was mapped into a 102.2 kb genomic region, and this region contained 12 genes. By RNA-seq data analysis, genome sequence comparison and functional validation through ectopic expression in Arabidopsis thaliana, Glyma.05G208300(described as GmEGL3), which is a basic helix-loop-helix(bHLH) transcription factor in plants, emerged as the most likely confirmed gene in qIF05-1. These long InDels can be used as a type of complementary genetic method for QTL fine mapping, and they can facilitate genetic studies and molecular-assisted selection breeding in soybean.展开更多
Geniposide,the principal active iridoid glucoside ingredient in Fructus gardeniae used in numerous traditional Chinese clinical prescriptions,has been shown to cause herbal hepatotoxicity because of its glycone metabo...Geniposide,the principal active iridoid glucoside ingredient in Fructus gardeniae used in numerous traditional Chinese clinical prescriptions,has been shown to cause herbal hepatotoxicity because of its glycone metabolite genipin.This study explored the role of gut microbiota in alleviating geniposide hepatotoxicity with isoflavones in soy products.Metabolic profiling using ultra high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UHPLC-Q/TOF-MS)revealed two metabolic pathways and six main forms of geniposides in vivo.Enzyme inhibitor experiments have shown that isoflavones alter geniposide metabolism by mediating specific enzymes,includingβ-glucosidase(β-GC)and sulfotransferase(SULT),in an established pseudo-sterile rat model.Isoflavones pretreatment by gavage for three weeks optimized the structure of the gut microbiota was linked to the regulation of key metabolic enzymes.Furthermore,experiments involving fecal microbiota transplantation(FMT)established the direct contribution of the gut microbiota to the regulation of enzyme activities and geniposide metabolism.This study demonstrated that isoflavones in soy products regulated the metabolic enzymes of geniposode dependent on gut microbiota,especially Lactobacillus spp.,which was further verified in our clinical trials analyzed using 16S ribosomal RNA(rRNA)and metagenomic sequencing,thus regulating geniposide metabolism.Furthermore,as dominant beneficial bacterium,Lactobacillus spp.were discovered to be promising microbial targets for the better management of geniposide hepatotoxicity.These findings provide valuable insights for the prevention and intervention of drug-induced liver injury.展开更多
3-(4-Chlorophenyl)-7-[2-(piperazin-1-yl)ethoxy]-4H-chromen-4-one(CPEO-43)is a derivative of soybean isoflavone(SI),synthesized by introducing a chlorine atom and a piperazine group into the structure of natural SI.In ...3-(4-Chlorophenyl)-7-[2-(piperazin-1-yl)ethoxy]-4H-chromen-4-one(CPEO-43)is a derivative of soybean isoflavone(SI),synthesized by introducing a chlorine atom and a piperazine group into the structure of natural SI.In vitro experiments have demonstrated that CPEO-43 exhibits a notable inhibitory effect on both A549 cells and HCT116 cells.For the further development and utilization of CPEO-43,this study aims to establish and validate a liquid chromatography-tandem mass spectrometry(LC-MS/MS)quantitative analysis method for the pharmacokinetic study of CPEO-43.Normal rats were intragastrically administered different doses(2,6,and 20 mg/kg)of CPEO-43,and blood was taken from the ocular venous plexus at different time points.The blood concentration of CPEO-43 at different time points was determined using LC-MS/MS technology,and the pharmacokinetic parameters of the compound were calculated using the pharmacokinetic software DAS.The results indicated that the established LC-MS/MS method complies with the standards for bioanalytical method validation in the Chinese Pharmacopoeia(CHP)and can be applied to the pharmacokinetic study of CPEO-43.The pharmacokinetic software DAS non-compartmental model was successfully used to calculate the C_(max),T_(max),t_(1/2),AUC_(0-∞),MRTs,CL,and Vd pharmacokinetic parameters of the compound at different doses.The results were as follows:62.0±10.5,222.0±28.7,and 1384.5±376.4 ng/mL;8.5±1.2,6.0±0.0,and 11.0±6.2 h;15.6,15.0,and 18.5 h;1517.8±317.0,5328.7±864.4,and 45556.3±22735.6 ng·h/mL;17.8±1.2,17.7±0.8,and 20.0±3.2 h;1370.3±305.9,1153.5±205.6,and 505.3±179.8 mL/kg/h;30843.0±7458.0,24344.0±5237.0,and 13950.3±5996.9 mL/kg.These characteristics are of great significance for understanding the in vivo process of the drug,formulating dosing regimens,and evaluating the safety and efficacy of the drug.展开更多
Identifying a potential dietary non-pharmacological treatment to prevent cerebrovascular damage in Alzheimer's disease is crucial for alleviating cognitive decline in older adults and enhancing quality of life.Thi...Identifying a potential dietary non-pharmacological treatment to prevent cerebrovascular damage in Alzheimer's disease is crucial for alleviating cognitive decline in older adults and enhancing quality of life.This study featured the combined supplementation of soy lecithin(SL)and soy isoflavones(SIF),using in vivo animal models,in vitro vascular ring preparation,and cell studies to investigate the potential advantages and mechanisms of SL combined with SIF on cognitive function and cerebrovascular health from multiple perspectives.The results show that Aβcan significantly induce learning and memory impairment in rats,as well as pathological changes in brain blood vessels,exacerbating damage to cerebral vasodilation function and subsequently reducing cerebral blood flow in the brain.The above-mentioned phenomena induced by Aβcan be significantly improved by the combined intervention of SL and SIF.Further research has revealed that the combined intervention of SL and SIF can reverse the downregulation of the PI3K/PIP3/PDK-1/Akt/eNOS signaling pathway and phosphorylated protein expression induced by Aβin rat brain vascular tissues and bEND.3 cells.Silencing PDK-1 expression in bEND.3 cells showed that the upregulation effect of SL and SIF on Akt and eNOS disappeared.Here we find that prophylactically supplementation with SL in conjunction with SIF appears to effectively activate the PI3K/PIP3/PDK-1/Akt/e NOS pathway within cerebral vascular.This activation improves cerebrovascular vasodilation,offering potential protective effects for both cerebral vascular health and cognitive function.展开更多
Objective: To analyze the possible mechanism of Pueraria isoflavones inhibiting XOD and GLUT9 to reduce uric acid production and promote uric acid excretion. Methods: August 2021-April 2022, a total of forty SPF male ...Objective: To analyze the possible mechanism of Pueraria isoflavones inhibiting XOD and GLUT9 to reduce uric acid production and promote uric acid excretion. Methods: August 2021-April 2022, a total of forty SPF male Kunming mice were divided into the healthy group (carboxymethylcellulose sodium at a dose of 250 mg/kg), the model group (HUA mice were given carboxymethylcellulose sodium at a dose of 250 mg/kg), the low group (HUA mice were given pueraria isoflavone at a dose of 125 mg/kg), HUA mice were given pueraria isoflavones at a dose of 250 mg/kg once d frequency)and the high group (HUA mice were given pueraria isoflavones at a dose of 500 mg/kg once d frequency) dosage groups, with 8 mice in each group. The contents of uric acid (SUA), urea nitrogen (BUN) and creatinine (SCr) in serum and urine of each group were compared before and after intervention (30 d). Statistical differences of xanthine oxidase (XOD) and human glucose transporter 9(GLUT9), cy- clooxygenase- 2(COX-2), tumor necrosis factor (TNF-α) and interleukin-1 (IL-1β) contents in renal tissues of each group after intervention (30 d) were compared. Results: After intervention, kidney inflammatory factors (COX-2, TNF-α and IL-1β) in the model group were compared. Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were higher than those of healthy group(P<0.05). Renal inflammatory cytokines (COX-2, TNF-α and IL-1β) in low, medium and high dose groups;Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were lower than those of model group, and there were low > medium > high dose groups, the comparison between the two groups had statistical significance(P< 0.05). After intervention, the contents of 3 indicators in blood or urine(COX-2, TNF-α and IL-1β) all decreased compared with before intervention, and the differences in intra-group comparison were statistically significant (P<0.05). Conclusion: Pueraria isoflavones can treat HUA mice by inhibiting the expression of XOD and GLUT9, and then play a role in reducing uric acid pro- duction and promoting uric acid excretion, as well as alleviating the degree of disease inflammation.展开更多
Two new isoflavones (8, 3'-dihydroxy-7,4'-dimethoxyisoflavone, odoratin-7-0-[3-D-glu-copyranoside) and four known isoflavones (formononetin, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, calycosin, calycosi...Two new isoflavones (8, 3'-dihydroxy-7,4'-dimethoxyisoflavone, odoratin-7-0-[3-D-glu-copyranoside) and four known isoflavones (formononetin, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, calycosin, calycosin-7-0-(3-D-glucopyranoside) were isolated from the roots of Astragalus mem-branaceus (Fisch.) Bunge. Their structures were established by spectral analysis.展开更多
The high isoflavones content of soybeans is an important breeding goal due to the demonstrated benefits of isoflavones to human health and their association with plant resistance.In this study,quantitative trait loci(...The high isoflavones content of soybeans is an important breeding goal due to the demonstrated benefits of isoflavones to human health and their association with plant resistance.In this study,quantitative trait loci(QTL)mapping for soybean isoflavone aglycones,including daidzin,glycerin,and genistin,and total isoflavones content was performed in a population of 178 F2:6 recombinant inbred lines(RILs)which was generated from a cross between varieties Jinong 17 and Jinong 18.A genetic linkage map covering 1248 cM was constructed using the simple sequence repeat(SSR)molecular markers.The results revealed 22 isoflavone-related QTLs,5 for daidzin,7 for genistin,6 for glycerin,and 4 for total isoflavone content.Seven of these represented new QTLs.All QTL regions contained 6462 genes,of which 58 have been annotated for flavonoid synthesis.Using public databases,three candidate genes,namely Glyma.11G164400,Glyma.16G158400,and Glyma.19G217700,were subsequently identified.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)demonstrated that the three genes exhibited specific,high expression in soybean seeds and a positive correlation with flavonoid content.These findings might be helpful in the efforts to breed new soybean varieties with improved isoflavone composition and content.展开更多
[Objective]The aim of this study was to set up a high performance liquid chromatography for rapid determination of isoflavones from soybean and analyze the correlation between isofalvone content and protein or fat con...[Objective]The aim of this study was to set up a high performance liquid chromatography for rapid determination of isoflavones from soybean and analyze the correlation between isofalvone content and protein or fat content. [Method]The isoflavones were firstly extracted by 80% methanol and then hydrolyzed at 100 ℃. The chromatographic separation adopted a reversed-phase C18 analytical column with binary high-pressure gradient elution,while its analysis time was 25 min and column temperature was 40 ℃. The diode array detector was used for monitoring with wavelength of 260 nm. The correlation between isofalvone content and protein or fat content was analyzed by data processing system Origin 6.0. [Result]The high performance liquid chromatograph for determination of isoflavones from soybean was verified to be accurate and reliable by methodology. The isoflavones of 85 soybean lines cultivated in Jilin Province were determined,and the results primarily showed the characters and ranges of isoflavones from soybean lines cultivated in Jilin Province,while the isoflavone content of soybeans ranged from 2.29 to 4.89 mg/g,and the average content was 3.36 mg/g. The isoflavone content of 5 soybean lines exceeded 4 mg/g,while there was a remarkably negative correlation between isoflavone content and protein content,and there was no significant positive correlation between isoflavone content and fat content. [Conclusion]The isoflavone content of soybean lines cultivated in Jilin Province is higher,so it is feasible for breeding the soybean lines with high isoflavone content and fat contetnt.展开更多
The fragmentation behavior of isoflavones was studied using electrospray ionization-ion trap-time of flight mass spectrometry （ESI-IT-TOF-MSn）. It was found that the isoflavone glycoside bond was easily broken. The ...The fragmentation behavior of isoflavones was studied using electrospray ionization-ion trap-time of flight mass spectrometry （ESI-IT-TOF-MSn）. It was found that the isoflavone glycoside bond was easily broken. The fragmentation occurred mostly on the C-ring, and the fragment ions of A^1,3＋ produced by the RDA cracking will predict the hydroxylation replacement on A-ring or B-ring. In addition, four carbonyl groups on the C-ring were fragmented through neutral loss of 28 （-CO）. A and B-rings primarily lose substituents which including a neutral losses of 32 （-CH3OH）, 16 （-CH4）, or 16 （-O）, and 18 （-H2O）. A-ring in the presence of adjacent hydroxylation, also easily made to be a neutral losses of 28 （-CO） or 18 （-H20）. It is likewise common to see methoxy replaced with a neutral losses of 16 （-CH4） or 32 （-CHaOH） in B-ring, also the hydroxylation on benzene ring can occasionally results with the neutral loss of 28 （-CO）.展开更多
Two major isoflavone glycosides [calycosin 7-O-β-D-glucopyranoside （1） and ononin （2）] and their aglycones [calycosin （3） and formononetin （4）] were simultaneously quantified with HPLC/DAD method. Two unknown...Two major isoflavone glycosides [calycosin 7-O-β-D-glucopyranoside （1） and ononin （2）] and their aglycones [calycosin （3） and formononetin （4）] were simultaneously quantified with HPLC/DAD method. Two unknown compounds were identified as calycosin 7-O-β-D-glucopyranoside-6＇＂-O-malonate （U1） and formononetin 7-O-β-D-glucopymnoside-6＇＂-O-malonate （U2）, respectively, with LC/MS^n. Raw Radix astragli were shown to have higher contents of isoflavone glycosides （1, 2）, but lower contents of aglycones （3, 4） than the processed herbal materials. After being moistened with water and stored up for 24 h at 35 ℃, the glycosides and their m_alonates were almost completely transformed to their corresponding aglycones. The different contents of the isoflavone glycosides and their aglycones in raw and processed Radix astragali materials might be due to enzymolysis of the glycosides during processing.展开更多
In this paper, we proposed a novel resolution criterion（improved calibrated normalized resolution product, r＊＇） to evaluate separation quality of fingerprints. By comparing with the calibrated normalized resolutio...In this paper, we proposed a novel resolution criterion（improved calibrated normalized resolution product, r＊＇） to evaluate separation quality of fingerprints. By comparing with the calibrated normalized resolution product（r＊） and the hierarchical chromatographic response function（HCRF）, the validity of this criterion was demonstrated by experimental chromatograms. The soy isoflavone extract was selected as the analytical object. The initial and end percentages of methanol and elution time affecting gradient elution were tested by orthogonal design. The final optimized conditions were as follows. It was detected by UV absorbance at 254 nm, column temperature was maintained at 36 oC, solvent A was 0.1%（v/v） acetic acid, solvent B was methanol, gradient elution was from 34% to 65% B in a linear gradient in 25 min, and the flow-rate was set at 1.0 m L/min. In addition, the main ingredients of the soy isoflavone extract were confirmed by LC-ESI/MS.展开更多
The genetic analysis of soybean isoflavone content and its components were carried out based on the NC Ⅱ matingdesign in eight soybean varieties. The results showed that the isoflavone contents and its components of ...The genetic analysis of soybean isoflavone content and its components were carried out based on the NC Ⅱ matingdesign in eight soybean varieties. The results showed that the isoflavone contents and its components of soybean seedare quite differences among the tested materials, the contents of isoflavone and daidzein are controlled not only byadditive effects and but also by non-additive effects, while the content of genistin is dominated by non-additive effects,and genistein, glycitin and daidzin are mainly controlled by additive effects. There are significant differences in thecontents of isoflavone and its components among the combinations derived from different parents. Results also indicatedthat the tested traits are negatively heterosis except for the contens of daidzein and daidzin are positively heterosis basedon the data of the GCA and SCA in average heterosis values. In this research we have a suggestion that soybean varietywith high isoflavone should be used as one of the parents in the breeding program, and it is the best choice that thecombinations crossed between two high isoflavone varieties or a high variety and a low one.展开更多
A series of novel N-[α-(isoflavone-7-O-)acetyl] amino acid methyl esters were prepared from the efficient and regioselective alkylation of isoflavones with chloroacetyl amino acid derivatives under mild condition.
Vilsmeier reagent formed from phthaloyl dichloride and DMF was found to be very effective for converting 2-hydroxyacetophenones, deoxybenzoins and dihydrochalcones into corresponding chromones, isoflavones and homoiso...Vilsmeier reagent formed from phthaloyl dichloride and DMF was found to be very effective for converting 2-hydroxyacetophenones, deoxybenzoins and dihydrochalcones into corresponding chromones, isoflavones and homoisoflavones with excellent yield. This method offers significant advantages such as efficiency and mild reaction conditions with shorter reaction time.展开更多
A new isoflavone glucoside, 3'-methoxy-5'-hydroxy-isoflavone-7-O-beta-D-glucoside, was isolated from the roots of Astragalus membranaceus and its structure was elucidated by spectroscopic data.
Objective:To clarify the effect of soy isoflavones on prevention of osteoporosis,and the effective dosage of soy isoflavones and its duration.Methods:Random control trials that investigated the association of soy isof...Objective:To clarify the effect of soy isoflavones on prevention of osteoporosis,and the effective dosage of soy isoflavones and its duration.Methods:Random control trials that investigated the association of soy isoflavones and osteoporosis were included in the meta-analysis by researching MEDLINE,EMBASE and the Chinese Biomedical Database up to October 2011.The RevMan software was used for all of the statistical analysis.Results:The present meta-analysis found that soy isoflavones significantly increased the bone mineral density by 54%and decreased the bone resorption marker urinary deoxypyridinoline(DPD) by 23%compared to baseline in women. Using random effects model,the effect of isoflavones on bone mineral density(BMD) regarding menopausal status and isoflavone dose revealed higher weighted mean difference changes were found in postmenopausal women and isoflavone dose above 75 mg/d.Subgroup analysis of trials with menopausal status,supplement type,isoflavone dose and intervention duration that used soy isoflavone extracts resulted in significant diflerent overall effect of DPD using by random effects model.Sensitivity analysis indicated that the effect of soy isoflavones on BMD and DPD was robust.Conclusions:The present meta-analysis reveals that soy isoflavone supplements significandy increase bone mineral density and decrease the bone,resorption marker urinary DPD.It shows no significant effect on bone formation markers serum bone alkaline phosphatase. The significant effect of soy isoflavones on BMD and urinary DPD is relative to menopausal status, supplement type,isoflavone dose and intervention duration.展开更多
It is important to determine the isoflavone components by high-performance liquid chromatography （HPLC） for the molecular assistant selection of isoflavone in soybean. Based on the standard samples of 12 isoflavone ...It is important to determine the isoflavone components by high-performance liquid chromatography （HPLC） for the molecular assistant selection of isoflavone in soybean. Based on the standard samples of 12 isoflavone components, the isoflavone components were analyzed using the determination of absorbance peaks method by HPLC. The results showed that there were different maximum ultraviolet （UV） absorbance for the aglycones of daidzein, glycitein, and genistein, which were at 250, 257, and 260 nm, respectively. A linear gradient elution of acetonitrile （13-30%） containing 0. 1% acetic acid as a mobile phase was applied on a YMC-C18 column at 35℃. The 12 isoflavone components were determined using the UV detector by HPLC. We concluded that this is a rapid and precise method which adapted to determine the large numbers of samples with microanalysis.展开更多
CGRP receptor（CLR） is a B class GPCR that functions only when combined with RAMPs. CLR/RAMP1 has been regarded as a promising target for migraine treatment, as its antagonists have been proved to be effective recent...CGRP receptor（CLR） is a B class GPCR that functions only when combined with RAMPs. CLR/RAMP1 has been regarded as a promising target for migraine treatment, as its antagonists have been proved to be effective recently. In the present study we designed and synthesized small molecular antagonists against CLR/RAMP1, resulting in a novel type of structure with acceptable high potency. The molecules were designed via virtual screening. Afterwards, a series of modification were conducted on the hit compounds, resulting in compound 8 as the best scored compound in docking, which was further validated in vitro by cell-based functional assay.展开更多
Soybean （Glycine max L. Merr.） is the world＇s foremost source of edible plant oil and proteins, meantime, the biologically active secondary metabolites such as saponins and isoflavones are benefit to human health. ...Soybean （Glycine max L. Merr.） is the world＇s foremost source of edible plant oil and proteins, meantime, the biologically active secondary metabolites such as saponins and isoflavones are benefit to human health. The objective of this study was to identify quantitative trait loci （QTL） and epistatic interactions associated with isoflavone, protein, and oil contents in soybean seeds. An F13 recombinant inbred line （RIL） comprising 474 lines was derived from a cross between Jindou 23 and Huibuzhi cultivars. SSR technique was employed for mapping of the QTLs. The QTLs for isoflavone, protein, and oil contents were analyzed and 23 QTLs were detected based on the constructed linkage map. Six QTLs for isoflavone content were localized in linkage groups J, N, D2, and G, eleven QTLs for oil content were localized in the linkage groups A1, A2, B2, C2, and D2, and six QTLs for protein content were localized in linkage groups B2, C2, G, and H1. The correlative analysis demonstrated that the isoflavone content had significant correlation with protein content, while significantly negative correlations was existed between oil and protein content, and significantly positive correlations was existed between protein and oil content. All these findings have laid an important basis for the marker assisted breeding in soybean. The phenotypic correlations of quantitative traits may be resulted from the correlation of the QTL controlling those traits.展开更多
文摘Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Iα-loop-helix Iβmotif which corresponds to helix I of other P 450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,theγ-domain.The structure reveals a homodimer in which theγ-domain of one molecule interacts with theβ-domain of another.The plane of heme group makes an angle of approximately 40°with the helix Iα-loop-helix Iβmotif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recognition.Phe-301,Ser-303 and Gly-305 from the helix Iα-loop-helix Iβmotif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.
基金supported by the China Agriculture Research System of MOF and MARA(CARS-04-PS12)the Research and Development Program in the Key-Areas of Guangdong Province,China(2020B020220008)the Guangdong Agricultural Research System,China(2023KJ136-03).
文摘Soybean seed isoflavones are a type of secondary metabolites that can provide health and nutrition benefits for humans. In our previous study, a stable quantitative trait locus(QTL) qIF05-1 controlling the seed isoflavone content in soybean was detected on chromosome(Chr.) 05 in a recombinant inbred line(RIL) population from a cross of Huachun 2×Wayao. In this study, the parental lines were re-sequenced using the Illumina Solexa System with deep coverage. A total of 63,099 polymorphic long insertions and deletions(InDels)(≥15 bp)were identified between the parents Huachun 2 and Wayao. The InDels were unevenly distributed on 20chromosomes of soybean, varying from 1,826 in Chr. 12 to 4,544 in Chr. 18. A total of 10,002 long InDels(15.85% of total) were located in genic regions, including 1,139 large-effect long InDels which resulted in truncated or elongated protein sequences. In the qIF05-1 region, 68 long InDels were detected between the two parents. Using a progeny recombination experiment and genotype analysis, the qIF05-1 locus was mapped into a 102.2 kb genomic region, and this region contained 12 genes. By RNA-seq data analysis, genome sequence comparison and functional validation through ectopic expression in Arabidopsis thaliana, Glyma.05G208300(described as GmEGL3), which is a basic helix-loop-helix(bHLH) transcription factor in plants, emerged as the most likely confirmed gene in qIF05-1. These long InDels can be used as a type of complementary genetic method for QTL fine mapping, and they can facilitate genetic studies and molecular-assisted selection breeding in soybean.
基金supported by the National Natural Science Foundation of China(82461160264,82474056,and 82104124)the grant from the Noncommunicable Chronic Diseases-National Science and Technology Major Project(2023ZD0502605)+2 种基金the Science and Technology Development Fund,Macao SAR(FDCT 0025/2021/A1)the Shanghai Municipal Health Commission(2022XD037)the Shanghai Magnolia Talent Plan Pujiang Project(23PJD113)。
文摘Geniposide,the principal active iridoid glucoside ingredient in Fructus gardeniae used in numerous traditional Chinese clinical prescriptions,has been shown to cause herbal hepatotoxicity because of its glycone metabolite genipin.This study explored the role of gut microbiota in alleviating geniposide hepatotoxicity with isoflavones in soy products.Metabolic profiling using ultra high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UHPLC-Q/TOF-MS)revealed two metabolic pathways and six main forms of geniposides in vivo.Enzyme inhibitor experiments have shown that isoflavones alter geniposide metabolism by mediating specific enzymes,includingβ-glucosidase(β-GC)and sulfotransferase(SULT),in an established pseudo-sterile rat model.Isoflavones pretreatment by gavage for three weeks optimized the structure of the gut microbiota was linked to the regulation of key metabolic enzymes.Furthermore,experiments involving fecal microbiota transplantation(FMT)established the direct contribution of the gut microbiota to the regulation of enzyme activities and geniposide metabolism.This study demonstrated that isoflavones in soy products regulated the metabolic enzymes of geniposode dependent on gut microbiota,especially Lactobacillus spp.,which was further verified in our clinical trials analyzed using 16S ribosomal RNA(rRNA)and metagenomic sequencing,thus regulating geniposide metabolism.Furthermore,as dominant beneficial bacterium,Lactobacillus spp.were discovered to be promising microbial targets for the better management of geniposide hepatotoxicity.These findings provide valuable insights for the prevention and intervention of drug-induced liver injury.
基金Scientific Research Projects of Hebei North University(Grant No.XJ2024022).
文摘3-(4-Chlorophenyl)-7-[2-(piperazin-1-yl)ethoxy]-4H-chromen-4-one(CPEO-43)is a derivative of soybean isoflavone(SI),synthesized by introducing a chlorine atom and a piperazine group into the structure of natural SI.In vitro experiments have demonstrated that CPEO-43 exhibits a notable inhibitory effect on both A549 cells and HCT116 cells.For the further development and utilization of CPEO-43,this study aims to establish and validate a liquid chromatography-tandem mass spectrometry(LC-MS/MS)quantitative analysis method for the pharmacokinetic study of CPEO-43.Normal rats were intragastrically administered different doses(2,6,and 20 mg/kg)of CPEO-43,and blood was taken from the ocular venous plexus at different time points.The blood concentration of CPEO-43 at different time points was determined using LC-MS/MS technology,and the pharmacokinetic parameters of the compound were calculated using the pharmacokinetic software DAS.The results indicated that the established LC-MS/MS method complies with the standards for bioanalytical method validation in the Chinese Pharmacopoeia(CHP)and can be applied to the pharmacokinetic study of CPEO-43.The pharmacokinetic software DAS non-compartmental model was successfully used to calculate the C_(max),T_(max),t_(1/2),AUC_(0-∞),MRTs,CL,and Vd pharmacokinetic parameters of the compound at different doses.The results were as follows:62.0±10.5,222.0±28.7,and 1384.5±376.4 ng/mL;8.5±1.2,6.0±0.0,and 11.0±6.2 h;15.6,15.0,and 18.5 h;1517.8±317.0,5328.7±864.4,and 45556.3±22735.6 ng·h/mL;17.8±1.2,17.7±0.8,and 20.0±3.2 h;1370.3±305.9,1153.5±205.6,and 505.3±179.8 mL/kg/h;30843.0±7458.0,24344.0±5237.0,and 13950.3±5996.9 mL/kg.These characteristics are of great significance for understanding the in vivo process of the drug,formulating dosing regimens,and evaluating the safety and efficacy of the drug.
基金supported by the National Natural Science Foundation of China(82273620,81302427)。
文摘Identifying a potential dietary non-pharmacological treatment to prevent cerebrovascular damage in Alzheimer's disease is crucial for alleviating cognitive decline in older adults and enhancing quality of life.This study featured the combined supplementation of soy lecithin(SL)and soy isoflavones(SIF),using in vivo animal models,in vitro vascular ring preparation,and cell studies to investigate the potential advantages and mechanisms of SL combined with SIF on cognitive function and cerebrovascular health from multiple perspectives.The results show that Aβcan significantly induce learning and memory impairment in rats,as well as pathological changes in brain blood vessels,exacerbating damage to cerebral vasodilation function and subsequently reducing cerebral blood flow in the brain.The above-mentioned phenomena induced by Aβcan be significantly improved by the combined intervention of SL and SIF.Further research has revealed that the combined intervention of SL and SIF can reverse the downregulation of the PI3K/PIP3/PDK-1/Akt/eNOS signaling pathway and phosphorylated protein expression induced by Aβin rat brain vascular tissues and bEND.3 cells.Silencing PDK-1 expression in bEND.3 cells showed that the upregulation effect of SL and SIF on Akt and eNOS disappeared.Here we find that prophylactically supplementation with SL in conjunction with SIF appears to effectively activate the PI3K/PIP3/PDK-1/Akt/e NOS pathway within cerebral vascular.This activation improves cerebrovascular vasodilation,offering potential protective effects for both cerebral vascular health and cognitive function.
基金National Innovation and Entrepreneurship Training Program for College Students(No.S202010823014)Hunan Provincial College Student Innovation Training Project,No.(2021)199(S202110823045)。
文摘Objective: To analyze the possible mechanism of Pueraria isoflavones inhibiting XOD and GLUT9 to reduce uric acid production and promote uric acid excretion. Methods: August 2021-April 2022, a total of forty SPF male Kunming mice were divided into the healthy group (carboxymethylcellulose sodium at a dose of 250 mg/kg), the model group (HUA mice were given carboxymethylcellulose sodium at a dose of 250 mg/kg), the low group (HUA mice were given pueraria isoflavone at a dose of 125 mg/kg), HUA mice were given pueraria isoflavones at a dose of 250 mg/kg once d frequency)and the high group (HUA mice were given pueraria isoflavones at a dose of 500 mg/kg once d frequency) dosage groups, with 8 mice in each group. The contents of uric acid (SUA), urea nitrogen (BUN) and creatinine (SCr) in serum and urine of each group were compared before and after intervention (30 d). Statistical differences of xanthine oxidase (XOD) and human glucose transporter 9(GLUT9), cy- clooxygenase- 2(COX-2), tumor necrosis factor (TNF-α) and interleukin-1 (IL-1β) contents in renal tissues of each group after intervention (30 d) were compared. Results: After intervention, kidney inflammatory factors (COX-2, TNF-α and IL-1β) in the model group were compared. Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were higher than those of healthy group(P<0.05). Renal inflammatory cytokines (COX-2, TNF-α and IL-1β) in low, medium and high dose groups;Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were lower than those of model group, and there were low > medium > high dose groups, the comparison between the two groups had statistical significance(P< 0.05). After intervention, the contents of 3 indicators in blood or urine(COX-2, TNF-α and IL-1β) all decreased compared with before intervention, and the differences in intra-group comparison were statistically significant (P<0.05). Conclusion: Pueraria isoflavones can treat HUA mice by inhibiting the expression of XOD and GLUT9, and then play a role in reducing uric acid pro- duction and promoting uric acid excretion, as well as alleviating the degree of disease inflammation.
基金This project is supported by National Center of New Drug Research,Science and Technology Committee and Educational Committee of Shanghai.
文摘Two new isoflavones (8, 3'-dihydroxy-7,4'-dimethoxyisoflavone, odoratin-7-0-[3-D-glu-copyranoside) and four known isoflavones (formononetin, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, calycosin, calycosin-7-0-(3-D-glucopyranoside) were isolated from the roots of Astragalus mem-branaceus (Fisch.) Bunge. Their structures were established by spectral analysis.
基金funded by the Key Research and Development Program of Science and Technology of Jilin Province(No.20220202011NC).
文摘The high isoflavones content of soybeans is an important breeding goal due to the demonstrated benefits of isoflavones to human health and their association with plant resistance.In this study,quantitative trait loci(QTL)mapping for soybean isoflavone aglycones,including daidzin,glycerin,and genistin,and total isoflavones content was performed in a population of 178 F2:6 recombinant inbred lines(RILs)which was generated from a cross between varieties Jinong 17 and Jinong 18.A genetic linkage map covering 1248 cM was constructed using the simple sequence repeat(SSR)molecular markers.The results revealed 22 isoflavone-related QTLs,5 for daidzin,7 for genistin,6 for glycerin,and 4 for total isoflavone content.Seven of these represented new QTLs.All QTL regions contained 6462 genes,of which 58 have been annotated for flavonoid synthesis.Using public databases,three candidate genes,namely Glyma.11G164400,Glyma.16G158400,and Glyma.19G217700,were subsequently identified.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)demonstrated that the three genes exhibited specific,high expression in soybean seeds and a positive correlation with flavonoid content.These findings might be helpful in the efforts to breed new soybean varieties with improved isoflavone composition and content.
文摘[Objective]The aim of this study was to set up a high performance liquid chromatography for rapid determination of isoflavones from soybean and analyze the correlation between isofalvone content and protein or fat content. [Method]The isoflavones were firstly extracted by 80% methanol and then hydrolyzed at 100 ℃. The chromatographic separation adopted a reversed-phase C18 analytical column with binary high-pressure gradient elution,while its analysis time was 25 min and column temperature was 40 ℃. The diode array detector was used for monitoring with wavelength of 260 nm. The correlation between isofalvone content and protein or fat content was analyzed by data processing system Origin 6.0. [Result]The high performance liquid chromatograph for determination of isoflavones from soybean was verified to be accurate and reliable by methodology. The isoflavones of 85 soybean lines cultivated in Jilin Province were determined,and the results primarily showed the characters and ranges of isoflavones from soybean lines cultivated in Jilin Province,while the isoflavone content of soybeans ranged from 2.29 to 4.89 mg/g,and the average content was 3.36 mg/g. The isoflavone content of 5 soybean lines exceeded 4 mg/g,while there was a remarkably negative correlation between isoflavone content and protein content,and there was no significant positive correlation between isoflavone content and fat content. [Conclusion]The isoflavone content of soybean lines cultivated in Jilin Province is higher,so it is feasible for breeding the soybean lines with high isoflavone content and fat contetnt.
基金Hong Kong Baptist University Faculty Research Foundation(Grant No.FRG2/12-13/058)the Key Program of Guangxi Natural Science Foundation of China(Grant No.2013-GXNSFBA019187)
文摘The fragmentation behavior of isoflavones was studied using electrospray ionization-ion trap-time of flight mass spectrometry （ESI-IT-TOF-MSn）. It was found that the isoflavone glycoside bond was easily broken. The fragmentation occurred mostly on the C-ring, and the fragment ions of A^1,3＋ produced by the RDA cracking will predict the hydroxylation replacement on A-ring or B-ring. In addition, four carbonyl groups on the C-ring were fragmented through neutral loss of 28 （-CO）. A and B-rings primarily lose substituents which including a neutral losses of 32 （-CH3OH）, 16 （-CH4）, or 16 （-O）, and 18 （-H2O）. A-ring in the presence of adjacent hydroxylation, also easily made to be a neutral losses of 28 （-CO） or 18 （-H20）. It is likewise common to see methoxy replaced with a neutral losses of 16 （-CH4） or 32 （-CHaOH） in B-ring, also the hydroxylation on benzene ring can occasionally results with the neutral loss of 28 （-CO）.
基金National Natural Science Foundation of China(Grant No.20432030 and 20742005).
文摘Two major isoflavone glycosides [calycosin 7-O-β-D-glucopyranoside （1） and ononin （2）] and their aglycones [calycosin （3） and formononetin （4）] were simultaneously quantified with HPLC/DAD method. Two unknown compounds were identified as calycosin 7-O-β-D-glucopyranoside-6＇＂-O-malonate （U1） and formononetin 7-O-β-D-glucopymnoside-6＇＂-O-malonate （U2）, respectively, with LC/MS^n. Raw Radix astragli were shown to have higher contents of isoflavone glycosides （1, 2）, but lower contents of aglycones （3, 4） than the processed herbal materials. After being moistened with water and stored up for 24 h at 35 ℃, the glycosides and their m_alonates were almost completely transformed to their corresponding aglycones. The different contents of the isoflavone glycosides and their aglycones in raw and processed Radix astragali materials might be due to enzymolysis of the glycosides during processing.
基金National Higher-Education Institution General Research and Development Funding(Grant No.JKP2011010)
文摘In this paper, we proposed a novel resolution criterion（improved calibrated normalized resolution product, r＊＇） to evaluate separation quality of fingerprints. By comparing with the calibrated normalized resolution product（r＊） and the hierarchical chromatographic response function（HCRF）, the validity of this criterion was demonstrated by experimental chromatograms. The soy isoflavone extract was selected as the analytical object. The initial and end percentages of methanol and elution time affecting gradient elution were tested by orthogonal design. The final optimized conditions were as follows. It was detected by UV absorbance at 254 nm, column temperature was maintained at 36 oC, solvent A was 0.1%（v/v） acetic acid, solvent B was methanol, gradient elution was from 34% to 65% B in a linear gradient in 25 min, and the flow-rate was set at 1.0 m L/min. In addition, the main ingredients of the soy isoflavone extract were confirmed by LC-ESI/MS.
基金This work was jointly spored by the grants from the Natural Science Foundation of Henan Province(0511030500)the National 863 Program of China(2004AA2111112)the N ational Natural Science Foundation of China(30490250).
文摘The genetic analysis of soybean isoflavone content and its components were carried out based on the NC Ⅱ matingdesign in eight soybean varieties. The results showed that the isoflavone contents and its components of soybean seedare quite differences among the tested materials, the contents of isoflavone and daidzein are controlled not only byadditive effects and but also by non-additive effects, while the content of genistin is dominated by non-additive effects,and genistein, glycitin and daidzin are mainly controlled by additive effects. There are significant differences in thecontents of isoflavone and its components among the combinations derived from different parents. Results also indicatedthat the tested traits are negatively heterosis except for the contens of daidzein and daidzin are positively heterosis basedon the data of the GCA and SCA in average heterosis values. In this research we have a suggestion that soybean varietywith high isoflavone should be used as one of the parents in the breeding program, and it is the best choice that thecombinations crossed between two high isoflavone varieties or a high variety and a low one.
文摘A series of novel N-[α-(isoflavone-7-O-)acetyl] amino acid methyl esters were prepared from the efficient and regioselective alkylation of isoflavones with chloroacetyl amino acid derivatives under mild condition.
文摘Vilsmeier reagent formed from phthaloyl dichloride and DMF was found to be very effective for converting 2-hydroxyacetophenones, deoxybenzoins and dihydrochalcones into corresponding chromones, isoflavones and homoisoflavones with excellent yield. This method offers significant advantages such as efficiency and mild reaction conditions with shorter reaction time.
文摘A new isoflavone glucoside, 3'-methoxy-5'-hydroxy-isoflavone-7-O-beta-D-glucoside, was isolated from the roots of Astragalus membranaceus and its structure was elucidated by spectroscopic data.
文摘Objective:To clarify the effect of soy isoflavones on prevention of osteoporosis,and the effective dosage of soy isoflavones and its duration.Methods:Random control trials that investigated the association of soy isoflavones and osteoporosis were included in the meta-analysis by researching MEDLINE,EMBASE and the Chinese Biomedical Database up to October 2011.The RevMan software was used for all of the statistical analysis.Results:The present meta-analysis found that soy isoflavones significantly increased the bone mineral density by 54%and decreased the bone resorption marker urinary deoxypyridinoline(DPD) by 23%compared to baseline in women. Using random effects model,the effect of isoflavones on bone mineral density(BMD) regarding menopausal status and isoflavone dose revealed higher weighted mean difference changes were found in postmenopausal women and isoflavone dose above 75 mg/d.Subgroup analysis of trials with menopausal status,supplement type,isoflavone dose and intervention duration that used soy isoflavone extracts resulted in significant diflerent overall effect of DPD using by random effects model.Sensitivity analysis indicated that the effect of soy isoflavones on BMD and DPD was robust.Conclusions:The present meta-analysis reveals that soy isoflavone supplements significandy increase bone mineral density and decrease the bone,resorption marker urinary DPD.It shows no significant effect on bone formation markers serum bone alkaline phosphatase. The significant effect of soy isoflavones on BMD and urinary DPD is relative to menopausal status, supplement type,isoflavone dose and intervention duration.
基金supported by grants from the National Transgenic Plants Program of China (2008ZX08004-003)the National High-Tech R&D Program (863Program, 2006AA100104)the National Natural Science Foundation of China (30000107)
文摘It is important to determine the isoflavone components by high-performance liquid chromatography （HPLC） for the molecular assistant selection of isoflavone in soybean. Based on the standard samples of 12 isoflavone components, the isoflavone components were analyzed using the determination of absorbance peaks method by HPLC. The results showed that there were different maximum ultraviolet （UV） absorbance for the aglycones of daidzein, glycitein, and genistein, which were at 250, 257, and 260 nm, respectively. A linear gradient elution of acetonitrile （13-30%） containing 0. 1% acetic acid as a mobile phase was applied on a YMC-C18 column at 35℃. The 12 isoflavone components were determined using the UV detector by HPLC. We concluded that this is a rapid and precise method which adapted to determine the large numbers of samples with microanalysis.
基金National Basic Research Program of China(Grant No.2012CB518000)Specialized Research Fund for the Doctoral Program of Higher Education of China(Grant No.20120001110010)
文摘CGRP receptor（CLR） is a B class GPCR that functions only when combined with RAMPs. CLR/RAMP1 has been regarded as a promising target for migraine treatment, as its antagonists have been proved to be effective recently. In the present study we designed and synthesized small molecular antagonists against CLR/RAMP1, resulting in a novel type of structure with acceptable high potency. The molecules were designed via virtual screening. Afterwards, a series of modification were conducted on the hit compounds, resulting in compound 8 as the best scored compound in docking, which was further validated in vitro by cell-based functional assay.
基金supported by the National Key Project for Transgenic Organisms Breeding,China(2008ZX08004-005,2009ZX08004-001B)the National 948 Project of China(2009-Z34,2010-G23)+3 种基金the National 973 Program of China(2008CB117005)the Henan Province key Scientific and Technological Project,China(102101110100)the Transformational Fund of Agricultural Scientific and Technological Achievements in China(2008D00010209)and the National Key Technologies R&D Program of China(2006BAD521B01-3)
文摘Soybean （Glycine max L. Merr.） is the world＇s foremost source of edible plant oil and proteins, meantime, the biologically active secondary metabolites such as saponins and isoflavones are benefit to human health. The objective of this study was to identify quantitative trait loci （QTL） and epistatic interactions associated with isoflavone, protein, and oil contents in soybean seeds. An F13 recombinant inbred line （RIL） comprising 474 lines was derived from a cross between Jindou 23 and Huibuzhi cultivars. SSR technique was employed for mapping of the QTLs. The QTLs for isoflavone, protein, and oil contents were analyzed and 23 QTLs were detected based on the constructed linkage map. Six QTLs for isoflavone content were localized in linkage groups J, N, D2, and G, eleven QTLs for oil content were localized in the linkage groups A1, A2, B2, C2, and D2, and six QTLs for protein content were localized in linkage groups B2, C2, G, and H1. The correlative analysis demonstrated that the isoflavone content had significant correlation with protein content, while significantly negative correlations was existed between oil and protein content, and significantly positive correlations was existed between protein and oil content. All these findings have laid an important basis for the marker assisted breeding in soybean. The phenotypic correlations of quantitative traits may be resulted from the correlation of the QTL controlling those traits.
