Mechanism of Active Ingredients of Astragalus Membranacus in Treating Heart Failure： A Review / 中国实验方剂学杂志

Heart failure is one of the main cardiovascular system diseases at present， and it is a clinical syndrome caused by changes in cardiac structure and function， resulting in impaired ejection function or ventricular filling. Therefore， heart failure has become the most important cardiovascular disease in the 21st century. In recent years， the incidence of heart failure is increasing， and the survival rate of patients with heart failure is very low. Traditional Chinese medicine has rich experience in preventing and treating heart failure. With the modernization of traditional Chinese medicine， more and more attention has been paid to the research， development， and application of active ingredients in traditional Chinese medicine. Traditional Chinese medicine has unique advantages in improving the heart function of patients with heart failure by treating multiple targets and multiple pathways through syndrome differentiation. Astragalus membranacus， a traditional Chinese medicine， is a kind of medicine that benefits Qi and blood circulation and removes evil spirits. It has the functions of improving myocardial energy metabolism and hemodynamics， protecting myocardial muscle， and promoting angiogenesis. Astragalus membranaceus is often used to treat patients with heart failure， yielding remarkable results. In recent years， it has been found that astragaloside， Astragalus polysaccharide， quercetin， calyx isoflavones， and other main active ingredients of Astragalus membranacus can improve cardiac function and treat heart failure by inhibiting inflammatory response， myocardial apoptosis， and myocardial fibrosis. This paper reviewed the research progress of the action and mechanism of the active ingredients of Astragalus membranacus in the treatment of heart failure by studying relevant literature， with a view to providing a reference for its further research， development， and application in the prevention and treatment of heart failure.

Biotransformed soybean cream as a new nutraceutical for skin care: collagen stimulation in vitro and ex vivo

Treatments that attenuate the effects of hypoestrogenism in menopausal women have been gaining visibility. This study investigated the skin response to a phytoestrogen-enriched cosmetic formulation created by incorporating a biotransformed soybean extract (BE) into a cream-like matrix. Collagen-I expression was analyzed both in vitro (fibroblast cells) and ex vivo (skin explants). The results revealed an increased amount of collagen-I both in fibroblasts and human skin when treated with BE and BE-incorporated cream. Also, this collagen-I overexpression was inhibited by PHTPP, indicating a dependence on estrogen hormone receptor beta (ERβ) signaling. Moreover, BE was not harmful to skin microbiota, showing a promising nutricosmetic potential. Thus, this work presented a fully functional cream-like formulation that was shown to be safe and effectively increase collagen-I levels both in vitro and ex vivo.

Soybean isoflavones alleviate cerebral ischemia/reperfusion injury in rats by inhibiting ferroptosis and inflammatory cascade reaction / 南方医科大学学报

OBJECTIVE@#To explore the mechanism that mediates the effect of soybean isoflavones (SI) against cerebral ischemia/reperfusion (I/R) injury in light of the regulation of regional cerebral blood flow (rCBF), ferroptosis, inflammatory response and blood-brain barrier (BBB) permeability.@*METHODS@#A total of 120 male SD rats were equally randomized into sham-operated group (Sham group), cerebral I/R injury group and SI pretreatment group (SI group). Focal cerebral I/R injury was induced in the latter two groups using a modified monofilament occlusion technique, and the intraoperative changes of real-time cerebral cortex blood flow were monitored using a laser Doppler flowmeter (LDF). The postoperative changes of cerebral pathological morphology and the ultrastructure of the neurons and the BBB were observed with optical and transmission electron microscopy. The neurological deficits of the rats was assessed, and the severities of cerebral infarction, brain edema and BBB disruption were quantified. The contents of Fe2+, GSH, MDA and MPO in the ischemic penumbra were determined with spectrophotometric tests. Serum levels of TNF-α and IL-1βwere analyzed using ELISA, and the expressions of GPX4, MMP-9 and occludin around the lesion were detected with Western blotting and immunohistochemistry.@*RESULTS@#The rCBF was sharply reduced in the rats in I/R group and SI group after successful insertion of the monofilament. Compared with those in Sham group, the rats in I/R group showed significantly increased neurological deficit scores, cerebral infarction volume, brain water content and Evans blue permeability (P < 0.01), decreased Fe2+ level, increased MDA level, decreased GSH content and GPX4 expression (P < 0.01), increased MPO content and serum levels of TNF-α and IL-1β (P < 0.01), increased MMP-9 expression and lowered occludin expression (P < 0.01). All these changes were significantly ameliorated in rats pretreated with IS prior to I/R injury (P < 0.05 or 0.01).@*CONCLUSION@#SI preconditioning reduces cerebral I/R injury in rats possibly by improving rCBF, inhibiting ferroptosis and inflammatory response and protecting the BBB.

Isoflavones from Sojae Semen Praeparatum Regulate Lipid Metabolism in Atherosclerotic Mice Through PPARγ/LXRα/ABCA1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo study the effect of isoflavones from Sojae Semen Praeparatum （ISSP） on lipid metabolism in atherosclerotic mice， and decipher the underlying mechanism via the peroxisome proliferator-activated receptor gamma/liver X receptor alpha/ATP-binding cassette transporter A1 （PPARγ/LXRα/ABCA1） signaling pathway. MethodFifty ApoE-/- mice were randomly assigned into the model group， western medicine （atorvastatin calcium， 3.03 mg·kg-1） group， and low-， medium-， and high-dose ISSP （2.5， 5， 10 mg·kg-1， respectively） groups， with 10 rats in each group. Atherosclerosis model mice were established by bilateral ovariectomy and feeding high-fat diet. Another 10 ApoE-/- mice receiving ovariectomy and high-fat diet were taken as the sham group. Some mice died of postoperative infection， and finally 6 mice were included in each group. One week after operation， each group was administrated with corresponding drugs or equivalent amount of normal saline. After 12 weeks， the levels of triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and non-esterified fatty acids （NEFAs） in serum and liver tissue were measured. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in serum were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining and oil red O staining were used for observation of aortic plaque formation and liver lipid deposition. The mRNA and protein levels of PPARγ， LXRα， ABCA1， and ATP-binding cassette transporter G1 （ABCG1） in liver were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the sham group， the modeling of atherosclerosis increased the aortic plaque area （P<0.01）， elevated the serum TC， TG， LDL-C， TNF-α， and IL-6 levels （P<0.01）， decreased the level of HDL-C （P<0.01）， increased the liver index （P<0.05） and the levels of TC， TG， and NEFAs in liver （P<0.01）， and caused obvious hepatic fat vacuoles and lipid deposition. In addition， the modeling down-regulated the mRNA levels of PPARγ， LXRα， ABCA1 in liver （P<0.05， P<0.01），and regulated the mRNA and protein levels of ABCG1（P<0.05， P<0.01）. Compared with the model group， atorvastatin calcium and middle-， high-dose ISSP reduced the serum TC， TG， LDL-C， TNF-α， and IL-6 levels （P<0.01）， decreased the liver index （P<0.01）， alleviated the liver fat vacuoles and lipid deposition， and increased the levels of TC， TG， and NEFAs in the liver （P<0.05， P<0.01）. Furthermore， they up-regulated the mRNA and protein levels of PPARγ， LXRα， ABCA1， and ABCG1 in the liver （P<0.05， P<0.01）. ConclusionISSP may regulate lipid metabolism through PPARγ/LXRα/ABCA1 signaling pathway to down-regulate the expression of inflammatory cytokines in serum and alleviate liver lipid deposition， thereby suppressing the formation of atherosclerotic plaque.

Relationship between soy isoflavone metabolites in urine and sexual development of adolescent girls / 中国学校卫生

Objective@#To explore the relationship between urinary soybean isoflavone metabolites and sexual development of adolescent girls.@*Methods@#Fifty girls of grade 5 from one primary school in Xingang City who met the inclusion criteria were selected. Sexual characteristic development was assessed and soybean isoflavone metabolites in urine was tested by ultra high speed liquid chromatography tandem mass spectrometry (UPLC MS/MS).@*Results@#Levels of dye xylinone in girls with different stages of sexual development (advanced, normal, and delayed groups) varied significantly[(186.7±24.1，171.8±22.8，155.3±21.6)μmol/mol] ( F =3.53， P <0.05), highest in advanced group. Urinary genistein was significantly higher in menarche group than that of non menarche group[(213.4±22.8，166.3±21.7)μmol/mol] ( t =2.16， P <0.05). Urinary genistein may be associated with early sexual development among adolescent girls.@*Conclusion@#High level of urine genistein may be related to the early sexual development of adolescent girls.

The influence of phytoestrogens or estrogens on the proliferation of the rat endocervical mucosa

SUMMARY INTRODUCTION Although estrogen therapy is widely used against post-menopausal symptoms, it can present adverse effects, including endometrial cancer. Soy isoflavones are considered a possible alternative to estrogen therapy. However, there are still concerns whether isoflavones exert trophic effects on the uterine cervix. OBJECTIVES To evaluate the histomorphometric and immunohistochemical alterations in the uterine cervix of ovariectomized rats treated with soy isoflavones (Iso). METHODS Fifteen adult Wistar rats were ovariectomized (Ovx) and divided into three groups: Group I (Ovx), administered with vehicle solution; Group II (OVX-Iso), administered with concentrated extract of Iso (150 mg/kg) by gavage; and Group III (OVX-E2), treated with 17β-estradiol (10 µg/kg), subcutaneously. After 30 days of treatments, the uterine cervix was fixed in 10% formaldehyde and processed for paraffin-embedding. Sections were stained with Hematoxylin and eosin for morphological and morphometric studies or subjected to immunohistochemistry for detections of Ki-67 and vascular endothelial growth factor-A (Vegf-A). The data obtained were subjected to statistical analysis (p ≤ 0.05). RESULTS We noted an atrophic uterine cervix in GI, whereas it was more voluminous in GII and even more voluminous in GIII. The thickness of the cervical mucosa was significantly higher in GIII, as compared to GI and GII. The cell proliferation (Ki-67) was significantly elevated in the estradiol and isoflavones treated groups, whereas Vegf-A immunoexpression was significantly higher in GIII, as compared to groups GII and GI. CONCLUSIONS Soy isoflavones cause less trophic and proliferative effects in the uterine cervix of rats as compared to estrogen.

RESUMO INTRODUÇÃO Embora a terapia estrogênica seja amplamente utilizada contra sintomas pós-menopausais, ela pode apresentar efeitos adversos, incluindo câncer de mama e endometrial. Assim, as isoflavonas da soja são consideradas uma alternativa possível à terapia estrogênica. No entanto, ainda há controvérsias se estes compostos exercem efeitos tróficos significativos no colo do útero. OBJETIVOS Avaliar as alterações histomorfométricas e imuno-histoquímicas no colo do útero de ratas ovariectomizadas tratadas com isoflavonas da soja (iso). MÉTODOS Quinze ratas Wistar adultas foram ovariectomizadas bilateralmente (Ovx) e separadas em três grupos: Grupo I (Ovx) - veículo (propilenoglicol); Grupo II (Ovx-Iso) - receberam extrato concentrado de Iso (150 mg/kg) e Grupo III (Ovx-E2) - tratado com 17β-estradiol (10 µg/kg); as soluções foram administradas via gavagem por 30 dias consecutivos. Posteriormente, os colos uterinos foram retirados, fixados em formaldeído a 10% tamponado e processados para inclusão em parafina. Cortes (4 µm) foram coradas com hematoxilina e eosina para estudo morfológico e morfométricos, enquanto outros foram submetidos à imuno-histoquímica para detecção de Ki-67 e do fator de crescimento endotelial vascular-A (Vegf-A). Os dados obtidos foram submetidos à análise estatística (p≤0,05). RESULTADOS Observamos a presença de colo uterino atrófico no GI (Ovx), sendo este mais volumoso no GII (Ovx+Iso) e ainda mais volumoso no GIII (Ovx+E2). A espessura da mucosa cervical foi significativamente maior no GIII (Ovx-E2), em comparação ao GI (Ovx) e ao GII (Ovx-Iso). A proliferação celular (Ki-67) foi significativamente mais elevada nos grupos tratados com estradiol e isoflavonas, enquanto a imunoexpressão de Vegf-A foi significativamente maior no GIII (Ovx-E2), em comparação ao GII (Ovx-Iso) e ao GI (Ovx-E2). CONCLUSÕES As isoflavonas da soja causam menos efeitos tróficos e proliferativos no colo do útero de ratas em comparação ao estrogênio.

Nutrikinetic study of fermented soybean paste (Cheonggukjang) isoflavones according to the Sasang typology

BACKGROUND/OBJECTIVES: In Oriental medicine, certain foods may be beneficial or detrimental based on an individual's constitution; however, the scientific basis for this theory is insufficient. The purpose of this study was to investigate the effect of body constitution, based on the Sasang type of Korean traditional medical classification system, on the bioavailability of soy isoflavones of Cheonggukjang, a quick-fermented soybean paste.SUBJECTS/METHODS: A pilot study was conducted on 48 healthy Korean men to evaluate the bioavailability of isoflavone after ingestion of food based on constitution types classified by the Sasang typology. The participants were classified into the Taeeumin (TE; n = 15), Soyangin (SY; n = 15), and Soeumin (SE; n = 18) groups. Each participant ingested 50 g of Cheonggukjang per 60 kg body weight. Thereafter, blood was collected, and the soy isoflavone metabolites were analyzed by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. Ntrikinetic analysis of individual isoflavone-derived metabolites was performed.RESULTS: Our nutrikinetic analysis identified 21 metabolites derived from isoflavones in the blood samples from 48 healthy Korean men (age range, 21-29 years). Significant differences were observed in the time to maximum concentration (T(max)) and elimination half-life (t(1/2)) for nine metabolites among the three groups. The T(max) and t(1/2) of the nine metabolites were higher in the SE group than in the other groups. Moreover, the absorption rates, as determined by the area under the plasma-level curve (AUC) values of intact isoflavone, were 5.3 and 9.4 times higher in the TE group than in the SY and SE groups, respectively. Additionally, the highest AUC values for phase I and II metabolites were observed in the TE group.CONCLUSIONS: These findings indicate that isoflavone bioavailability, following Cheonggukjang insgestion, is high in individuals with the TE constitution, and relatively lower in those with the SE and SY constitutions.

Urine Metabonomic Analysis of Interventions Effect of Soy Isoflavones on Rats Exposed to Di-(2-ethylhexyl) Phthalate / 生物医学与环境科学（英文）

Objective@#Di-(2-ethylhexyl) phthalate (DEHP) is a ubiquitous environmental contaminant. As an endocrine disruptor, it seriously threatens human health and ecological environmental safety. This study examines the impact of intervention with soybean isoflavones (SIF) on DEHP-induced toxicity using a metabonomics approach.@*Methods@#Rats were randomly divided into control (H), SIF-treated (A, 86 mg/kg body weight), DEHP-treated (B, 68 mg/kg), and SIF plus DEHP-treated (D) groups. Rats were given SIF and DEHP daily through diet and gavage, respectively. After 30 d of treatment, rat urine was tested using UPLC/MS with multivariate analysis. Metabolic changes were also evaluated using biochemical assays.@*Results@#Metabolomics analyses revealed that p-cresol glucuronide, methyl hippuric acid, N1-methyl-2-pyridone-5-carboxamide, lysophosphatidycholine [18:2 (9Z, 12Z)] {lysoPC [18:2 (9Z, 12Z)]}, lysoPC (16:0), xanthosine, undecanedioic acid, and N6-acetyl-l-lysine were present at significantly different levels in control and treatment groups.@*Conclusion@#SIF supplementation partially protects rats from DEHP-induced metabolic abnormalities by regulating fatty acid metabolism, antioxidant defense system, amino acid metabolism, and is also involved in the protection of mitochondria.

A reliable and rapid pharmacokinetic study of pueraria isoflavones using pueraria reference extractive substance in beagle plasma: Application to study of Yufeng Ningxin Tablets / 中草药·英文版

Objective: In order to evaluate the reliability and feasibility of pueraria reference extractive substance (RES) used in biological sample, the pharmacokinetics of 3′‑hydroxy puerarin (3′-HP), puerarin, 3′‑methoxy puerarin (3′-MP), and daidzein-8-C-apiosyl-(1-6)-glucoside (DAG) in beagle plasma following oral administration of Yufeng Ningxin Tablet were quantitated. Methods: A reliable and sensitive high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS) method developed with chromatographic separation was operated on a Merck C18 column, and acetonitrile-5 mmol/L ammonium was used as mobile phase in gradient elution. The plasma samples were deproteinized by acetone, detected by triple quadrupole mass spectrometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. The pharmacokinetic parameters were calculated by Winnonlin 4.1. Results: The calibration curves of the reference extractive substance and standard substance methods were linear over the ranges 0.0417–11.3309 µg/mL and 0.0394–10.0000 µg/mL. The intra-day and inter-day precision of the two methods at three concentrations were less than 13.63%, and the average recoveries of 3′-HP, puerarin, 3′-MP, and DAG were more than 70.67%. The RSD of the mean plasma concentrations of the analytes calculated by the two methods was less than 5%, and cos (ϑ) = =1.000. Among the analytes, puerarin showed the highest blood concentration [(940 ± 185) ng/mL] and the longest retention time [(5 ± 1) h] in the dog's bodies. Conclusion: Pueraria reference extractive substance can be seen as an alternative to the standard substance to overcome the scarcity of standard substance for the analysis of biological samples.

Phytochemicals and antioxidant activity of alcoholic/hydroalcoholic extract of Trifolium pratense / 中草药·英文版

Objective: Trifolium pratense has many healing properties, including fewer complications of menopause, cancer cell suppression, reducing blood glucose and lipids, as well as cardiovascular beneficial effects. The purpose of this study was to identify the phytochemical and mineral composition of T. pratense. Methods: Plant aerial parts were harvested and dried, and then hydroalcoholic and alcoholic extracts were prepared. Gas chromatography–mass spectrometry (GC–MS) analytical method was used to identify volatile compounds then liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) was used to identify polyphenols and the mineral elements were identify by inductively coupled plasma atomic emission spectrometer/ICP-AES and scanning electron microscope-energy-dispersive X-ray spectroscopy (SEM-EDS) methods. Total phenolic content (TPC) was determined based on colorimetric method, and total flavonoid content (TFC) was established based on the folin-chiocalteau reagent. Furthermore, two assays (DPPH and FRAP) were used to measure the antioxidant capacity of T. pratense ethanolic extract. Results: A total of 37 polyphenols and 107 peaks were identified by LC-ESI-MS analysis, and the GC/MS method also detected 21 volatile compounds, the most important of which were methylcyclopentane, dimethylpentanal and hexadecanol. A total of 18 mineral elements, including K, Mg, Al, Si, Zn, Ni, Cu, Se, Co, Fe, Mn, and Ca in the plant, were identified ICP-AES and SEM-EDS analysis. Conclusion: T. pratense has many therapeutic compounds such as polyphenol (isoflavone and flavonoids), volatile compounds, and essential mineral elements, which can be formulated purely and used in the pharmaceutical and traditional medicine industries.

Rapid Identification and Analysis of Chemical Components in Heartwood of Dalbergia cochinchinensis by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective::To rapidly identify and analysis the chemical constituents in the methanol extract of heartwood of Dalbergia cochinchinensis by ultra high performance liquid chromatography-quadrupole-time-of-flight high resolution mass spectrometry (UPLC-Q-TOF-MS/MS). Method::UPLC RRHD SB-C18 column (3.0 mm×100 mm, 1.8 μm) was used for chromatographic separation with acetonitrile-0.1% formic acid solution as the mobile phase for gradient elution (0-0.01 min, 5%B; 0.01-2 min, 5%-22%B; 2-28 min, 22%-35%B; 28-45 min, 35%-44%B; 45-55 min, 44%-100%B; 55-57 min, 100%B; 57-57.10 min, 100%~5%B) at a flow rate of 0.3 mL·min-1. The analytes were determined in negative ion mode with electrospray ionization (ESI) and data collection range of m/z 100-1 500. Result::A total of 101 chemical constituents were identified, including 22 flavonoids, 34 isoflavones, 15 neoflavonoids, 18 other flavonoids and 12 other components. Conclusion::UPLC-Q-TOF-MS/MS technique can quickly, accurately and comprehensively identify the chemical constituents in the heartwood of D. cochinchinensis. Isoflavones, flavonoids and neoflavonoids are the main chemical constituents in the heartwood of D. cochinchinensis, which is of great significance to reveal its internal material basis and provides experimental basis for this plant to be developed as a potential new resource of traditional Chinese medicine.

Rapid Identification on Chemical Components in Dalbergiae Odoriferae Lignum by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective::Dalbergiae Odoriferae Lignum is a rare traditional Chinese medicine material in China. However, there are many varieties of various sources and different qualities in the market at present. In order to further define the pharmacodynamic substance basis, electrospray time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) was used to rapidly analyze chemical constituents of methanol extract of Dalbergiae Odoriferae Lignum. Method::Chromatographic separation was performed on an UPLC RRHD SB-C18(3.0 mm×100 mm, 1.8 μm)for gradient elution, with mixtures of acetonitrile and 0.1%formic acid-water as mobile phases at a flow rate of 0.3 mL·min-1. The column temperature was maintained at 40 ℃. The data was collected in a negative ion mode with electro-spray ionization source(ESI). Result::According to molecular ion peaks and MS2 mass spectrometry characteristic fragment ions, Mass Bank databases, as well as the mass spectrometry information of reference substances and relevant literatures, a total of 83 constituents were identified, including 18 flavones, 31 isoflavones, 10 neoflavonoids, 9 isoflavanones, 7 other flavonoids and 8 other components. Conclusion::UPLC-Q-TOF-MS/MS can quickly, accurately and comprehensively identify chemical constituents in methanol extract of Dalbergiae Odoriferae Lignum, and isoflavones, flavones, neoflavonoids and isoflavanones are the main chemical constituents, which laid a foundation for the basic research of medicinal substances of Dalbergiae Odoriferae Lignum, and provided theoretical basis and technical support for the improvement of quality standards of Dalbergiae Odoriferae Lignum.

Research Advances on Processing Procedures，Chemical Constituents, Quality Analysis and Pharmacological Effect of Sojae Semen Germinatum / 中国实验方剂学杂志

Sojae Semen Germinatum （SSG） was firstly recorded in Shennong Bencaojing. It is a dry processed product which is germinated using mature seeds of Glycine max. It is neutral in nature and sweet flavor， and its functions are to relieve heat， clear away heat and remove dampness. SSG has a long history of being used both as food and medicine， but it was not enrolled in the Chinese Pharmacopoeia until 2010. Doctors of different dynasties had different views of its processing procedures， and thus the quality of its decoction pieces is inconsistent. This article systematically straightened out the records of SSG in ancient books and modern literature， and summarized and analyzed the processing procedures， chemical constituents， quality analysis and pharmacological effects of SSG. It found that SSG contains proteins， isoflavones， saponins and other components， analytical methods for detecting these components include ultraviolet spectrophotometry （UV）， thin-layer chromatography （TLC）， high performance liquid chromatography （HPLC）， etc. It has effect such as antioxidant， anti-inflammatory， anti-osteoporosis， improvement of menopausal syndrome， treatment of cardiovascular diseases， and processing will change the type and content of its chemical components. Therefore， it is necessary to dig out active constituents of SSG， explore those changes in chemical constituents and pharmacological effect during the period of its primary and subsequent processing， and explore its action mechanism. This paper can provide the theoretical basis for standardized processing procedure， modern quality control and clinical application of SSG.

Isoflavones' effects on pharmacokinetic profiles of main iridoids from Gardeniae Fructus in rats / 药物分析学报

Gardeniae Fructus (GF) and Semen Sojae Praeparatum (SSP) are both medicine food homologies and widely used in Chinese clinical prescriptions together. The research investigated the pharmacokinetics of four iridoids in normal rats and isolfavones-fed rats, which were administered with isolfavones from SSP for 7, 14, 21 and 28 consecutive days. A validated LC-MS/MS method was developed for determining shanzhiside, genipin-1-gentiobioside, geniposide and their metabolite genipin in rat plasma. Plasma samples were pretreated by solid-phase extraction using paeoniflorin as the internal standard. The chromatographic separation was performed on a Waters Atlantis T3 (4.6 mm × 150 mm, 3μm) column using a gradient mobile phase consisting of acetonitril and water (containing 0.06％acetic acid). The mass detection was under the multiple reaction monitoring (MRM) mode via polarity switching between negative and positive ionization modes. The calibration curves exhibited good linearity (r>0.997) for all components. The lower limit of quantitation was in the range of 1-10 ng/mL. The intra-day and inter-day precisions (RSD) at three different levels were both less than 12.2％ and the accuracies (RE) ranged from -10.1％ to 16.4％. The extraction recovery of them ranged from 53.8％ to 99.7％. Pharmacokinetic results indicated the bioavailability of three iridoid glycosides and the metabolite, genipin in normal rats was higher than that in rats exposed to isoflavones. With the longer time of administration of iso-flavones, plasma concentrations of iridoids decreased, while genipin sulfate, the phase II metabolite of genposide and genipin-1-gentiobioside, appeared the rising exposure. The pharmacokinetic profiles of main iridoids from GF were altered by isoflavones.

Establishment of HPLC Fingerprint and Content Determination of 5 Kinds of Isoflavone Components in Sojae Semen Nigrum / 中国药房

OBJECTIVE：To establ ish the fingerprint of Sojae Semen Nigrum and content determination method of 5 kinds of isoflavones，so as to provide reference for controlling its quality better. METHODS ：HPLC method was adopted to establish the fingerprint and detect the contents of 5 kinds of isoflavones. The determination was performed on Phenomenex C 18 column with mobile phase consisted of acetonitrile- 0.12％ formic acid solution （gradient elution ）at the flow rate of 1 mL/min. The detection wavelength was set at 260 nm；the column temperature was 30 ℃ and sample size was 10 μL. Using daidzin as reference，HPLC fingerprints of 12 batches of samples were determined. The similarity of 12 batches of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System （2012A） to confirm common peak. Cluster analysis and principal component analysis were performed by using SPSS 20.0 software and SIMCA 13.0 software. RESULTS ：There were 19 common peaks in HPLC fingerprints of 12 batches of samples ，the similarity of which was higher than 0.94. Totally 5 components were identified，such as daidzin ，glycitin，genistin，daidzein，genistein. Cluster analysis showed that 12 batches of Sojae Semen Nigrum were clustered into 2 categories，i.e. S 1-S3 clustered into one category ，and S 4-S12 clustered into the other category. By principal component analysis ，the contribution rates of two principle components were 53.261％ and 40.715％；accumulative contribution rate was 93.976％. The linear range of above 5 components were 5.97-191.00 µg/mL（r＝0.999 9），1.05-33.46 µg/mL（r＝0.999 9）， 8.93-285.61 µg/mL（r＝0.999 5），0.82-26.33 µg/mL（r＝0.999 9），0.93-29.64 µg/mL（r＝0.999 7），respectively. The limits of quantitation were 0.881 1，0.611 6，0.078 6，0.243 3，0.511 6 μg/mL，respectively. The limits of detection were 0.264 3，0.244 7， 0.021 4，0.124 8，0.106 7 μg/mL，respectively. RSDs of precision ，stability，reproducibility and durability tests were all lower than 5％. Recoveries were 95.15％-96.56％（RSD＝0.51％，n＝6），98.52％-103.45％（RSD＝1.88％，n＝6），95.37％-97.91％ （RSD＝0.95％，n＝6），99.75％-102.00％（RSD＝0.78％，n＝6），100.26％-103.65％（RSD＝1.21％，n＝6）. Among 12 batches of Sojae Semen Nigrum ，the contents of above 5 components were 0.178 3-0.265 9，0.021 7-0.096 2，0.288 5-0.597 2，0.014 1- 0.058 8，0.012 9-0.082 9 mg/g. CONCLUSIONS ：Established HPLC fingerprint and content determination method of 5 kinds of isoflavones can be used for quality control of Sojea Semen Nigrum. The Isoflavone components are similar ，but the contents are different among Sojae Semen Nigrum from different producing areas.

Extraction, characterization and microencapsulation of isoflavones from soybean molasses / Extração, caracterização e microencapsulação de isoflavonas do melaço de soja

ABSTRACT: This study aimed to extract isoflavones from soybean molasses with different solvents, furthermore, the selected extract, which showed overall desirable characteristics was selected to evaluate the potentials of different encapsulating agents. The encapsulating agents employed for the study included 18% Maltodextrin DE20 (T1), 18% Hi-maize (T2), and a mixture of equal proportions of 9% Maltodextrin DE20 and 9% of Hi-maize (T3). Solvents such as 80% ethanol and methanol, and grain alcohol in varying different concentrations of 50 and 80% were used for the studies. The best solvent for the extraction of phenolics and total isoflavones was 50% cereal alcohol, this extract also presented higher antioxidant activity. Evaluation of the encapsulating agents revealed that 18% Hi-maize with inlet air of 130 °C was best suited for the encapsulation of isoflavones. The ORAC method showed that microcapsules with the 18% Hi-maize encapsulating agent also had higher antioxidant activity.

RESUMO: Este estudo teve como objetivo extrair isoflavonas do melaço de soja com diferentes solventes, além disso, o extrato selecionado com as características desejáveis ​​em geral foi selecionado para avaliar os potenciais de diferentes agentes encapsulantes. Os agentes encapsulantes foram: 18% de maltodextrina DE20 (T1), 18% Hi-maize (T2) e uma mistura em proporções iguais de 9 % de maltodextrina DE20 e 9% de Hi-maize (T3). Foram testados os solventes, etanol e metanol a 80% e álcool de cereais a 50 e 80%. O melhor solvente para a extração de fenólicos e isoflavonas totais foi o álcool de cereais a 50%, sendo que este extrato também apresentou maior atividade antioxidante. Entre os agentes encapsulantes testados, 18% Hi-maize com ar de entrada de 130 °C mostrou ser o melhor para encapsular as isoflavonas. O método ORAC mostrou que as microcápsulas com o agente encapsulante Hi-maize a 18% também apresentaram maior atividade antioxidante.

Solid state fermentation to obtain vegetable products bio-enriched with isoflavone aglycones using lactic cultures / Fermentación en sustrato sólido utilizando cultivos lácticos para la obtención de productos vegetales bioenriquecidos con isoflavonas agliconas

The consumption of soybean isoflavones (IS) is associated with several beneficial properties on human health. Some lactic acid bacteria possess ß-glucosidase enzyme, that allows to obtain the active form of IS (aglycone). The solid state fermentation (SSF) has received great attention in the last years in order to obtain several valuable compounds. SSF, using soybean as substrate and Lactobacillus rhamnosus CRL 981 as starter, was studied in the present work. Sucrose was added into soybean paste to study the effect on the behavior of the selected strain. The development of L. rhamnosus CRL 981 through pH and recount measures, sugar intake, organic acid production, ß-glucosidase activity and IS conversion were analyzed. No significant differences in growth and acidity were observed between soybean pastes with and without sucrose added, but the production of lactic acid was higher in the latter paste. The ß-glucosidase activity was detected in both pastes and the complete hydrolysis of IS at 12 h of fermentation was observed. Also, this strain was able to increase the free amino acids in soybean paste. SSF, using soybean as substrate and L. rhamnosus CRL 981 as starter culture, is an alternative process to obtain a soybean product bio-enriched in active IS with attractive nutritional characteristics.

El consumo de isoflavonas de soja (IS) está asociado a diversos beneficios para la salud humana. Ciertas bacterias lácticas poseen la enzima ß-glucosidasa, que permite obtener la forma bioactiva (agliconas) de las IS. La fermentación en sustrato sólido (FSS) ha recibido gran atención en los últimos anos debido a sus numerosas ventajas, y permite la obtención de productos con valor agregado. En el presente trabajo se estudió la FSS utilizando soja como sustrato y Lactobacillus rhamnosus CRL981 como cultivo iniciador. Con el fin de estudiar el efecto de una fuente de carbono externa sobre el comportamiento de la cepa seleccionada, se adicionó sacarosa a la pasta de soja. Se evaluó el crecimiento de L. rhamnosus CRL 981 a través de medidas de pH y recuento en placa. Además, se analizó el consumo de azúcares, producción de ácidos orgánicos, actividad ß-glucosidasa y conversión de IS. No se observaron diferencias significativas en el crecimiento y acidez entre las pastas de soja sin adición de sacarosa y con ella, sin embargo, la producción de ácido láctico fue mayor en esta última. La actividad de ß-glucosidasa se detectó en ambas pastas y se observó la hidrólisis completa de IS a las 12 h de fermentación. Además, esta cepa fue capaz de aumentar los aminoácidos libres en la pasta de soja. La FSS, utilizando soja como sustrato y L. rhamnosus CRL 981 como cultivo iniciador, es un proceso alternativo para obtener un producto de soja bioenriquecido en IS bioactivas con características nutricionales atractivas.

Nutrigenomics as a tool in the prevention of lipotoxicity: the case of soy protein

Abstract Obesity is associated with an increase of several metabolic disorders leading to the development of diseases such as type 2 diabetes and cardiovascular disease. This is due in part to the ectopic accumulation of triglycerides in organs that are non-adipose tissues, leading to lipotoxicity. Particularly, in the liver, the accumulation of lipids, mainly of triglycerides, leads to the formation of fatty liver. The accumulation of lipids in skeletal muscle and pancreas associates with insulin resistance and a decrease in insulin secretion, respectively. In addition, it has been suggested that dysbiosis of the gut microbiota can contribute to the process of lipid accumulation in non-adipose tissues, especially in the liver. The aim of the present review is to highlight the mechanisms associated with the development of lipotoxicity, and how with the advances in nutrigenomics, it is now possible to understand the molecular mechanisms by which some nutrients can attenuate the ectopic accumulation of triglycerides in non-adipose tissues. Particularly, we emphasize research conducted on the molecular mechanisms of action of soy protein and some of its isoflavones, and how these can reduce lipotoxicity by preventing the accumulation of lipids in the liver, skeletal muscle, and pancreas, as well as their role on the gut microbiota to attenuate the development of fatty liver. Thus, nutrigenomics is opening new dietary strategies based on several functional foods that can be used to ameliorate the pathologies associated with lipotoxicity.

Effect of Isoflavones and Genistein on Glucose Metabolism in Peri- and Post-Menopausal Women: An Overview of Meta-Analysis

The purpose of the present overview of meta-analysis is to summarize and critically assess the effect of isoflavones and genistein on glucose metabolism among the peri- and post-menopausal women. Two independent authors searched the databases of MEDLINE, Scopus and Cochrane Library for meta-analysis. Three databases were searched from inception to January 2018. Methodological quality of each meta-analysis of randomized controlled trials was evaluated using the AMSTAR (a measurement tool used to assess systematic reviews). Four meta-analyses were included to the current overview. Fasting insulin levels and homeostatic model assessment of insulin resistance (HOMA-IR) values were significantly lower in peri-menopausal and postmenopausal. Two meta-analyses showed that treatment with isoflavones could not alter fasting blood glucose. However, one meta-analysis depicted that isoflavones significantly improved blood glucose levels in non-Asian postmenopausal women. Treatment with genistein could have significant beneficial effects on fasting insulin, blood glucose and HOMA-IR in comparison to the control group. Regardless of the population, the treatment with genistein is effective in improving fasting insulin, HOMA-IR and glucose levels. Nevertheless, the high heterogeneity among studies and poor methodology of reviews made it difficult to draw a definite conclusion on the positive impacts of soy on glucose metabolism.

Effects of Isoflavone Supplementation on Lipid Profiles and Antioxidant Enzyme Activities in Growing Rats Fed High Fat Diet

The purpose of this study was to investigate the effects of isoflavone on serum lipids and antioxidant enzymes activities in growing rats fed high lard diet. Twenty four female Sprague-Dawley rats (body weight 50–60 g) were divided into three groups, control, high fat (HF, lard 200 g/kg diet) and high fat + isoflavone (HFI, lard 200 g/kg diet + isoflavone 310.9 mg/kg diet) for 4 weeks. The results of study indicated that body weight gain was not different by isoflavone diet. Mean intake was significantly lower in HF group and HFI group than control group. Food efficiency ratio was significantly higher in HF group and HFI group than control group. The level of serum triglyceride and total cholesterol were significantly lower in HFI group than control group and HF group. The level of high-density lipoprotein cholesterol, was significantly higher in control group than HF group and HFI group. The level of low-density lipoprotein cholesterol was not significantly different by experimental diets, but atherogenic index (AI) was significantly lower in control group and HFI group than HF group. Contents of total cholesterol and triglyceride in liver tissues were found to be insignificant. The concentration of lipid peroxidation, malondialdehyde was significantly lower in control groups and HFI group than HF group. And antioxidant enzymes in liver tissue were not significantly different by lard and isoflavone supplemented diets. In conclusion, it seems possible that isoflavone supplemented high fat diet may produce positive results on level of serum triglyceride, serum total cholesterol, AI and concentration of malondialdyhyde.