Differentiation of human adipose-derived stem cells into neuron-like cells by Radix Angelicae Sinensis

Human adipose tissues are an ideal source of stem cells. It is important to find inducers that can safely and effectively differentiate stem cells into functional neurons for clinical use. In this study, we investigate the use of Radix Angelicae Sinensis as an inducer of neuronal differentiation. Primary human adipose-derived stem cells were obtained from adult subcutaneous fatty tissue, then pre-induced with 10% Radix Angelicae Sinensis injection for 24 hours, and incubated in serum-free Dulbecco＇s modified Eagle＇s medium/Nutrient Mixture F-12 containing 40% Radix Angelicae Sinensis to induce its differentiation into neuron-like cells. Butylated hydroxyanisole, a common in- ducer for neuronal differentiation, was used as the control. After human adipose-derived stem cells differentiated into neuron-like cells under the induction of Radix Angelicae Sinensis for 24 hours, the positive expression of neuron-specific enolase was lower than that of the butylated hydroxyani- sole-induced group, and the expression of glial fibrillary acidic protein was negative. Alter they were induced for 48 hours, the positive expression of neuron specific enolase in human adipose-derived stem cells was significantly higher than that of the butylated hydroxyanisole-induced group. Our experimental findings indicate that Radix Angelicae Sinensis can induce human adipose-derived stem cell differentiation into neuron-like cells and produce less cytotoxicity.

Progress of Radix Astragali and Radix Angelicae Sinensis in the treatment of idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis(IPF)is a chronic,progressive,fibrotic interstitial lung disease.Current treatment options for IPF are limited.Radix Astragali(RA)and Radix Angelicae Sinensis(RAS),according to 5:1 ratio composed of Danggui Buxue decoction(DGBXD),which have played an essential role in the treatment of IPF.This article reviewed the experimental research,clinical research,and progress of RA and RAS(DGBXD)treating IPF to provide a deeper scientific basis for the future experimental research and clinical research.

Study on mechanism and molecular docking verification of Angelicae Sinensis Radix and Astragali Radix in treating ischemic stroke

Background:Traditional Chinese medicine(TCM)has been shown to be effective in treating ischemic stroke(IS),and the combination of Angelicae Sinensis Radix(ASR)and Astragali Radix(AR)is a core TCM prescription that is widely acknowledged for its efficacy in IS treatment.This study utilized network pharmacology methods to explore the molecular mechanisms underlying the therapeutic effects of Angelicae Sinensis Radix and Astragali Radix in IS treatment,with preliminary validation conducted through molecular docking.Methods:Information on the structure,targets,main biological functions,and pathways of the active components in Angelicae Sinensis Radix and Astragali Radix was collected using databases such as PubChem,PharmMapper,UniProt,and GeneCards.The results were visualized using software such as Cytoscape 3.6.1,Ledock,and pymol.Results:We retrieved 20 active components and 149 targets associated with the compatibility of Angelicae Sinensis Radix and Astragali Radix from various databases,and GeneCards database was used to search 3350 IS-related gene targets,including 78 key targets of Angelicae Sinensis Radix and Astragali Radix for the treatment of IS.Enrichment analysis of these 78 targets using gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)revealed the involvement of 48 GO terms in the treatment of IS,mainly in biological processes such as metabolism,biological regulation,and stress response.The composition of biological devices such as supercavitary membrane,cell fluid,and extracellular space was also involved.The biological functions mainly included protein binding,ion binding,hydrolytic enzyme activity,and others.The identified pathways were estrogen signaling pathway,mitogen-activated protein kinase(MAPK)signaling pathway,PI3K-AKT signaling pathway,RAP1 signaling pathway,P53 signaling pathway,PPAR signaling pathway,FOXO signaling pathway,RAS signaling pathway,prolactin signaling pathway,HIF-1 signaling pathway,and TNF signaling pathway.Molecular docking analysis showed that the 17 key active components of Angelicae Sinensis Radix and Astragali Radix had strong binding activity with 13 IS key targets.Conclusion:Through the application of network pharmacology methods,it was found that the use of Angelicae Sinensis Radix and Astragali Radix for treating ischemic stroke mainly targets the MAPK and PI3K-AKT signaling pathways,involving several crucial compounds and genes.Nevertheless,additional in vitro and in vivo studies are needed to verify these findings.

Evaluation of Angelicae sinensis radix as a promising treatmentoption for hyperlipidemia based on network pharmacology

Background: Angelicae sinensis radix has been widely applied in traditional Chinese medicine while little isexplored in its potential mechanism. This study aims to elucidate the effective components and defattingmechanism based on network pharmacology. Methods: Traditional Chinese Medicine Systems PharmacologyDatabase and Analysis Platform was screened to collect the possible active ingredients and their CAS and SMILESwas searched in Pubchem, which further used for reverse molecular docking in Swiss Target Prediction database toobtain potential targets. Hyperlipidemia-related molecules were obtained from GeneCards database, and thepredicted targets of Angelicae sinensis radix for hyperlipidemia treatment were selected by Wayne diagram. Formechanism analysis, the protein-protein interactions were constructed with String, the Gene Oncology enrichmentanalysis and Kyoto Encyclopedia of Genes and Genomes analysis were conducted in DAVID. Results: Usingnetwork-based systems biology analysis, we predicted that 5 active ingredients in Angelicae sinensis radix hasantilipemic effects with 71 potential targets. Through Gene Oncology and Kyoto Encyclopedia of Genes andGenomes analysis, we found that the related signaling pathways mainly involved in arachidonic acid metabolism,and regulation of lipolysis in adipocytes. The related genes are ALOX5, CYP2C19, EPHX2, PTGS1, PTGS2,ADRB1, and ADRB3. Conclusion: Angelicae sinensis radix may alleviate hyperlipidemia through arachidonic acidmetabolism, and regulation of lipolysis in adipocytes. ALOX5, CYP2C19, EPHX2, PTGS1, PTGS2, ADRB1, andADRB3 may be new targets for treatment.

Potential mechanism of Astragali radix-Angelicae sinensis radix in the treatment of spinal cord injury based on network pharmacology and molecular docking

Objective:Using network pharmacology and molecular docking technology to explore the possible mechanism of Huangqi(Astragali radix)-Danggui(Angelicae sinensis radix)on the treatment of spinal cord injury.Methods:The active components and the targets related to Astragali radix-Angelicae sinensis radix were screened out on the Traditional Chinese Medicine Systems Pharmacology database.Genes of spinal cord injury were searched by Genecards and the Online Mendelian Inheritance in Man databases.The intersection targets between herbs and diseases were obtained through online Venn diagrams.A components-targets-pathways network was established on Cytoscape 3.8.1 software.The STRING database was used to construct the intersection protein interaction network and screen out core targets.Gene Ontology biological processes and enrichment analysis based on the Kyoto Encyclopedia of Genes and Genes of intersection proteins were performed via DAVID database.Finally,the molecular docking with key components and core targets were performed in AutoDock software.Results:The 22 chemical components including quercetin,kaempferol were collected from Astragali radix-Angelicae sinensis radix.It acts on 110 targets,and interleukin-6,tumor necrosis factor,mitogen-activated protein kinase,tumor antigen p53 were considered as the major targets.50 pathways like Interleukin-17 signaling pathway,tumor necrosis factor signaling pathway and mitogen-activated protein kinase signaling pathway participate in biological processes such as positive transcription regulation and lipopolysacchanide response.The molecular docking revealed that the core targets had stronger binding activity with its corresponding active components.Conclusion:Astragali radix-Angelicae sinensis radix has the characteristics of multi-component,multi-target,and multi-pathway effects in treating spinal cord injury.Its potential mechanism may be related to preventing inflammation,improving microcirculation,inhibiting neuronal apoptosis,protecting damaged nerve cells and promoting nerve repair and regeneration.

Effect of co-existent components in CO2 supercritical fluid extract of Angelica Sinensis Radix on metabolism of Z-ligustilide after oral administration in rats

Objective:To establish a basis for Angelica Sinensis Radix(ASR)as a dietary supplement for colorectal cancer chemoprevention,the effect of co-existent components in supercritical fluid extract(SFE)of ASR on the pharmacokinetics of Z-ligustilide after oral administration was investigated in vitro and in vivo.Methods:Incubation in gastrointestinal contents and incubation in rat liver tissue homogenates post-mitochondrial supernatant(PMS)experiments were used to study changes in the levels of Z-ligustilide in vitro.Results:Within 4 hours,the level of Z-ligustilide in SFE declined at a slower rate than in its pure form.Clearance of Z-ligustilide after administration in its pure form was significantly slower than that of SFE of ASR(CL,0.96±0.16 mL·min/kg versus 1.24±0.21 mL·min/kg P<0.05;AUC,243.37±16.84 versus 176.69±12.59 mg·min/L).Conclusion:These phenomena may be attributed to the interactions between the co-existent components in SFE of ASR and Z-ligustilide enhancing the stability of Z-ligustilide.These results suggest that the bioavailability of Z-ligustilide in SFE of ASR is improved.However,stabilization of plasma concentration was not sustained,so that the efficacy of active components could not be maintained.Thus,further processing of SFE of ASR is required.

Hepatic metabolomics combined with network pharmacology to reveal the correlation between the anti-depression effect and nourishing blood effect of Angelicae Sinensis Radix

Angelicae Sinensis Radix(AS)is reproted to exert anti-depression effect(ADE)and nourishing blood effect(NBE)in a rat model of depression.The correlation between the two therapeutic effects and its underlying mechanisms deserves further study.The current study is designed to explore the underlying mechanisms of correlation between the ADE and NBE of AS based on hepatic metabonomics,network pharmacology and molecular docking.According to metabolomics analysis,30 metabolites involved in 11 metabolic pathways were identified as the potential metabolites for depression.Furthermore,principal component analysis and correlation analysis showed that glutathione,sphinganine,and ornithine were related to pharmacodynamics indicators including behavioral indicators and hematological indicators,indicating that metabolic pathways such as sphingolipid metabolism were involved in the ADE and NBE of AS.Then,a target-pathway network of depression and blood deficiency syndrome was constructed by network pharmacology analysis,where a total of 107 pathways were collected.Moreover,37 active components obtained from Ultra Performance Liquid Chromatography-Triple-Time of Flight Mass Spectrometer(UPLC-Triple-TOF/MS)in AS extract that passed the filtering criteria were used for network pharmacology,where 46 targets were associated with the ADE and NBE of AS.Pathway enrichment analysis further indicated the involvement of sphingolipid metabolism in the ADE and NBE of AS.Molecular docking analysis indciated that E-ligustilide in AS extract exhibited strong binding activity with target proteins(PIK3CA and PIK3CD)in sphingolipid metabolism.Further analysis by Western blot verified that AS regulated the expression of PIK3CA and PIK3CD on sphingolipid metabolism.Our results demonstrated that sphingolipid metabolic pathway was the core mechanism of the correlation between the ADE and NBE of AS.

Mechanisms exploration of Angelicae Sinensis Radix and Ligusticum Chuanxiong Rhizoma herb-pair for liver fibrosis prevention based on network pharmacology and experimental pharmacologylogy

Angelicae Sinensis Radix(Danggui)and Ligusticum Chuanxiong Rhizoma(Chuan Xiong)herb-pair(DC)have been frequently used in Traditional Chinese medicine(TCM)prescriptions for hundreds of years to prevent vascular diseases and alleviate pain.However,the mechanism of DC herb-pair in the prevention of liver fibrosis development was still unclear.In the present study,the effects and mechanisms of DC herb-pair on liver fibrosis were examined using network pharmacology and mouse fibrotic model.Based on the network pharmacological analysis of 13 bioactive ingredients found in DC,a total of 46 targets and 71 pathways related to anti-fibrosis effects were obtained,which was associated with mitogen-activated protein kinase(MAPK)signal pathway,hepatic inflammation and fibrotic response.Furthermore,this hypothesis was verified using carbon tetrachloride(CCl_4)-induced fibrosis model.Measurement of liver functional enzyme activities and histopathological examination showed that DC dramatically reduced bile acid levels,inflammatory cell infiltration and collagen deposition caused by CCl_4.The increased expression of liver fibrosis markers,such as collagen 1,fibronectin,α-smooth muscle actin(α-SMA)and transforming growth factor-β(TGF-β),and inflammatory factors,such as chemokine(C-C motif)ligand 2(MCP-1),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and IL-6 in fibrotic mice were significantly downregulated by DC herb-pair through regulation of extracellular signal-regulated kinase 1/2(ERK1/2)-protein kinase B(AKT)signaling pathways.Collectively,these results suggest that DC prevents the development of liver fibrosis by inhibiting collagen deposition,decreasing inflammatory reactions and bile acid accumulation,which provides insights into the mechanisms of herbpair in improving liver fibrosis.

基于GC-MS和网络药理学探讨当归-肉桂药对的抗炎作用机制

对当归(Angelica sinensis Radix)-肉桂(Cinnamomi cortex)药对化学成分进行鉴定,并结合网络药理学预测其抗炎作用机制。采用GC-MS(气相色谱-质谱)分析当归、肉桂混合物挥发性成分,再通过TCMSP数据库和Swiss Target Prediction数据库预测其活性成分的潜在靶点;通过TTD数据库、GeneCards数据库得到炎症相关基因靶点;利用Cytoscape 3.7.2软件构建药物-成分-靶点网络图;采用String平台、Cytoscape 3.7.2软件构建靶点PPI网络,并筛选出关键靶点;采用R 3.6.1软件中的Clsuter Profiler程序包对潜在作用靶点进行基因本体(GO)功能富集及KEGG通路富集分析。结果表明,从当归-肉桂药对中鉴定出35种挥发性成分,占总挥发性成分的70.53%。并从中筛选得到33个活性化合物、35个关键靶点、1296个生物过程、155条信号通路。根据网络药理学结果预测表明,当归-肉桂药对可能通过β-石竹烯、α-律草烯、α-蒎烯等活性成分及TNF、IL-6、IL-1β等关键靶点来调控脂质和动脉粥样硬化、人巨细胞病毒感染、IL-17等多个信号通路,从而协同发挥抗炎作用。

Danggui Buxue decoction promotes proliferation and differentiation of Caco-2 cells and antagonizes obesity by stimulating apolipoprotein-IV expression

Background:Chinese medicine has been proposed as a novel approach to the prevention of metabolic disorders such as obesity.Danggui Buxue decoction,a decoction prepared from Huangqi(Astragali Radix)and Danggui(Angelicae Sinensis Radix),has been used to nourish vitality and enhance blood circulation in traditional Chinese medicine.However,the effect of Danggui Buxue decoction on obesity is still primarily unknown.Methods:Cell proliferation,differentiation,and apolipoprotein-IV transcription were investigated to explore the function of Danggui Buxue decoction by methyl thiazolyl tetrazolium assay,alkaline phosphatase assay,and luciferase assay,respectively.Results:Danggui Buxue decoction promoted cell growth by up to 15%(P=0.034)and induced cell differentiation by up to 38%(P=0.006)at 0.3 mg/mL.Moreover,Danggui Buxue decoction enhanced the transcription of apolipoprotein-IV by 2.4 times(P=0.027),activating its promoter by 28.9%(P=0.031)at 0.3 mg/mL.In addition,Danggui Buxue decoction functioned in a dose-dependent manner.Conclusion:These results suggest that Danggui Buxue decoction promotes cell differentiation by enhancing apolipoprotein-IV transcription and alkaline phosphatase activity,and it may also have a potential anti-obesity effect because apolipoprotein-IV transcription is closely related to the reduction of food intake.

DNA metabarcoding analysis of fungal community on surface of four root herbs

Objective: Angelicae Sinensis Radix(ASR, Danggui in Chinese), Cistanches Herba(CH, Roucongrong in Chinese), Ginseng Radix et Rhizoma(PG, Renshen in Chinese), and Panacis Quinquefolii Radix(PQ,Xiyangshen in Chinese), widely used as medicine and dietary supplement around the world, are susceptible to fungal and mycotoxin contamination. In this study, we aim to analyze their fungal community by DNA metabarcoding.Methods: A total of 12 root samples were collected from three main production areas in China. The samples were divided into four groups based on herb species, including ASR, CH, PG, and PQ groups. The fungal community on the surface of four root groups was investigated through DNA metabarcoding via targeting the internal transcribed spacer 2 region(ITS2).Results: All the 12 samples were detected with fungal contamination. Rhizopus(13.04%-74.03%),Aspergillus(1.76%-23.92%), and Fusarium(0.26%-15.27%) were the predominant genera. Ten important fungi were identified at the species level, including two potential toxigenic fungi(Penicillium citrinum and P. oxalicum) and eight human pathogenic fungi(Alternaria infectoria, Candida sake, Hyphopichia burtonii, Malassezia globosa, M. restricta, Rhizopus arrhizus, Rhodotorula mucilaginosa, and Ochroconis tshawytschae). Fungal community in ASR and CH groups was significantly different from other groups,while fungal community in PG and PQ groups was relatively similar.Conclusion: DNA metabarcoding revealed the fungal community in four important root herbs. This study provided an important reference for preventing root herbs against fungal and mycotoxin contamination.

Qualitative Analysis of A Sulfur-fumigated Chinese Herbal Medicine by Comprehensive Two-Dimensional Gas Chromatography and High-Resolution Time of Flight MassSpectrometry Using Colorized Fuzzy Difference Data Processing

To investigate the chemical transformation of volatile compounds in sulfur-fumigated Radix Angelicae Sinensis. A comprehensive two-dimensional gas chromatography (GCxGC) and high-resolution time-of-flight mass spectrometry (HR-TOF/MS) with colorized fuzzy difference (CFD) method was used to investigate the effect of sulfur-fumigation on the volatile components from Radix Angelicae Sinensis. Twenty-five compounds that were found in sun-dried samples disappeared in sulfur-fumigated samples. Seventeen volatile components including two sulfur-containing compounds were newly generated for the first time in volatile oils of sulfur-fumigated Radix Angelicae Sinensis. The strategy can be successfully applied to rapidly and holistically discriminate sun-dried and sulfur-fumigated Radix Angelicae Sinensis. GCxGC-HR-TOF/MS based CFD is a powerful and feasible approach for the global quality evaluation of Radix Angelicae Sinensis as well as other herbal medicines.

Investigation on the Mode of Action of the Traditional Chinese Medical Prescription-Yiqihuoxue Formula, an Effective Extravasation Treatment for Cerebral Vascular Microemboli in ApoE-/- mice

Objective: The objective of this study was to investigate the mechanisms underlying anti-embolism and extravasational effects of traditional Chinese medical prescription YiqiHuoxue(YQHX) formula in ApoE-/-mice with cerebral vascular microemboli. Materials and Methods: An ApoE-/-mice model with microemboli was developed by infusing fluorescently labeled heterologous fibrin-rich microparticles into the internal carotid artery of ApoE -/-gene knockout male mice through the common carotid artery. Before microemboli injection, the animals were randomly divided into four groups of 10 animals, treated daily for 6 weeks by intragastric administration: The ApoE-/-control group(physiological saline, 0.2 mL/10 g/d), YQHX group(0.2 ml/10 g/d), clopidogrel group(3 mg/kg/d), and atorvastatin group(3 mg/kg/d);a further group was constituted of normal male C57 BL/6 J mice(with the same genetic background as ApoE-/-mice;normal control group;no treatment;microemboli injection). The mice in each microemboli group were divided into three subgroups, the 2-h, 24-h, and 72-h subgroups, corresponding to the time after microemboli injection. Two hours(or 24 h or 72 h) after microemboli injection, the changes in aortic intima and brain tissue were analyzed by histopathology, the amounts of fluorescent emboli being measured by fluorescence microscopy image analysis. Comparison points included the microemboli induced loss of aorta functions and pathological changes, atherosclerotic plaque, brain ultrastructure and functions, and embolus extravasation. Results: Loss of aorta functions and adverse pathological changes, atherosclerotic plaque, serious damage in brain ultrastructure and functions, and reduced thrombus elimination were obviously serious in microemboli injected ApoE-/-mice. These symptoms were significantly relieved by the YQHX pretreatment:(i) the ratio of thrombus accumulation was increased with a significant decrease in thrombus extravasation in ApoE-/-mice, while YQHX induced an increased thrombus extravasation;(ii) the degree of aortic intimal thickening and brain tissue structural disorders were significantly increased in ApoE-/-mice, but overtly inhibited in the YQHX group;(iii) YQHX restored cell viability and homeostasis in the brain;(iv) YQHX regulated the expression of pro-and anti-inflammatory cytokines in the aorta;and(v) YQHX reduced cortical nerve nuclei pyknosis, edema, liquefaction, and necrosis induced by brain hypoxia, especially in the 24 h and 72 h groups. Conclusions: These findings indicate that the protective effects of YQHX on the brain against microemboli-induced injury may be attributed to the activation of extravasation mechanisms, which are involved in the cerebrovascular injury pathway and constitutively important in the progression of ischemic stroke.