Analysis of principal isoflavone glycosides and aglycones in Radix astragali

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.

Molecular mechanism of Radix astragali on improvement of insulin sensitivity of SD rats treated with low dose dexamethasone

Aim To reveal the main active components and the action mechanisms of Radix astragali on insulin sensitivity improvement, we have investigated the effects of polysaccharide portion and saponin portion of Radix astragali extracts on blood biochemical indices and related gene expression of dexamethasone-induced SD rats. Methods SD rats （6 per group） received 2 μg/day subcutaneous dexamethasone for 4 weeks plus same dose （10 g material/kg） of polysaccharide or saponin extracts of Radix astragali. Blood samples, kidney tissues and epididymal fat pads were taken at the end of the experiment. Serum triglyceride （TG）, total cholesterol （TC）, low density lipoprotein cholesterol （LDLC）, high density lipoprotein cholesterol （HDLC）, glucose （GLU） and insulin （INS） levels were measured, respectively, mRNA levels of angiotensinogen in kidney, adiponectin and leptin as well as TNF-α in epididymal fats were determined by RT-PCR assay using GAPDH gene as an internal control. Results Both of polysaccharide and saponin extracts of Radix astragali exhibited positive effects in reducing serum triglycerides, glucose, and insulin levels of dexamethasone-induced SD rats. The saponin group showed more improvements on quantitive insulin sensitivity check index （QUICKI） than the polysaccharide group did. Both of the extracts down-regulated kidney angiotensinogen and fat TNF-α mRNA levels while they were simultaneously up-regulating fat adiponectin and leptin mRNA levels. No significant difference was found between actions of the two extracts. Conclusion Both of polysaccharide and saponin extracts of Radix astragali can improve insulin sensitivity. This action might be closely related to down-regulation of angiotensinogen, TNF-α and up-regulation of adiponectin and leptin expression. The results partly explained the improvement of type Ⅱ diabetes and diabetic nephropathy by Radix astragali. The similar actions of the two crude extracts suggest that unknown key active compounds might exist in both and remain to be discovered.

Protective effect of Radix Astragali injection on immune organs of rats with obstructive jaundice and its mechanism

AIM: To observe the protective effect of Radix Astragali injection on immune organs (lymph nodes, spleen and thymus) of rats with obstructive jaundice (OJ) and its mechanism. METHODS: SD rats were randomly divided into sham-operation group, model control group and Radix Astragali treatment group. On days 7, 14, 21 and 28 after operation, mortality rate of rats, pathological changes in immune organs, expression levels of Bax and nuclear factor (NF)-κB p65 proteins, apoptosis indexes and serum tumor necrosis factor (TNF)-α level in spleen and thymus were observed, respectively.RESULTS: Compared to model control group, the number of dead OJ rats in Radix Astragali treatment group decreased (P > 0.05). The TNF-α level (27.62 ± 12.61 vs 29.55 ± 18.02, 24.61 ± 9.09 vs 31.52 ± 10.95) on days 7 and 21, the pathological severity score for spleen [0.0 (0.0) vs 0.0 (2.0) on days 7 and 14 and for lymph nodes [0.0 (1.0) vs 1.0 (2.0), 1.0 (0.0) vs 2.0 (1.0)] on days 21 and 28, the product staining intensity and positive rate of Bax protein in spleen [0.0 (0.0) vs 1.0 (2.0), 0.0 (1.0) vs 2.0 (1.5) and thymus [0.0 (0.0) vs 1.0 (2.0), 0.0 (1.0) vs 2.0 (1.5)] on days 14 and 28, the apoptotic indexes [0.0 (0.0) vs 0.0 (0.01)] in spleen and thymus [0.0 (0.0) vs 0.0 (0.01) on days 14 and 21 were significantly lower in Radix Astragali treatment group than in model control group (P < 0.05). CONCLUSION: Radix Astragali has protective effects on immune organs of OJ rats by relieving the pathological changes in immune organs, reducing TNF-α level and inhibiting Bax expression and apoptosis in spleen and thymus.

Protective Effects of Radix Astragali against Anoxic Damages to in vitro Cultured Neurons

To study the protective effects of radix astragali against anoxic damages to in vitro cultured neurons in rats, NaCN was used to develop a hypoxic model of in vitro cultured neurons from newborn rat cerebral cortex. The cellular morphology, A value (cell survival number) and effluxes of lactate dehydrogenase(LDH) and K+ from cells were measured in the radix astragali group and the control group respectively. After 48 h of anoxia, A value was decreased from 0. 325± 0. 031 before anoxia to 0. 145± 0. 011, the effluxes of LDH and K+ were increased from 65. 80± 2. 90 U/L and 5. 23 ± 0. 11mmol/L before anoxia to 148. 80± 8. 40 U/L and 7. 31 ± 0. 18 mmol/L, respectively. It was found that in the anoxic circumstance in the Radix astragali group, the mophological changes were mild, the effluxes of LDH and K+ were decreased and A value increased as compared with those in the control group. It was suggested that Radix astragali could protect the cultured rat neurons against anoxic damages in the anoxic circumstance.

Predicting the grades of Astragali radix using mass spectrometrybased metabolomics and machine learning

Astragali radix(AR,the dried root of Astragalus)is a popular herbal remedy in both China and the United States.The commercially available AR is commonly classified into premium graded(PG)and ungraded(UG)ones only according to the appearance.To uncover novel sensitive and specific markers for AR grading,we took the integrated mass spectrometry-based untargeted and targeted metabolomics approaches to characterize chemical features of PG and UG samples in a discovery set(n=16 batches).A series of five differential compounds were screened out by univariate statistical analysis,including arginine,calycosin,ononin,formononetin,and astragalosideⅣ,most of which were observed to be accumulated in PG samples except for astragalosideⅣ.Then,we performed machine learning on the quantification data of five compounds and constructed a logistic regression prediction model.Finally,the external validation in an independent validation set of AR(n=20 batches)verified that the five compounds,as well as the model,had strong capability to distinguish the two grades of AR,with the prediction accuracy>90%.Our findings present a panel of meaningful candidate markers that would significantly catalyze the innovation in AR grading.

Simultaneous separation and determination of four main isoflavonoids in Astragali Radix by an isocratic LC/ESI-MS method

A simple, reliable and rapid isocratic liquid chromatography(LC)-mass spectrometric detection(MS) coupled with electrospray ionization(ESI) method for simultaneous separation and determination of calycosin-7-O-β-D-glucoside, ononin, calycosin and formonometin in Astragali Radix was developed. After the samples were extracted with ethanol, the optimum separation conditions for these analytes were achieved using water and acetonitrile(70:30, v/v) containing 0.2%(v/v) acetic acid as a mobile phase and a 2.0 mm×150 mm Hypersil-Keystone C18 column. Selective ion monitoring(SIM) mode and [M+H]+ ions at m/z 447, 431, 285 and 269 were used for quantitative analysis of four main active components above mentioned. The calibration curves were linear in the range of 0.4-175.0 μg/mL for calycosin-7-O-β-D-glucoside, 0.2-146.0 μg/m L for ononin, 0.4-210.0 μg/mL for calycosin and 0.5-217.0 μg/mL for formonetion, respectively. The limits of quantification(LOQ) and detection(LOD) were 0.4 μg/mL and 0.08 μg/m L for calycosin-7-O-β-D-glucoside, 0.2 μg/mL and 0.06 μg/m L for ononin, 0.4 μg/mL and 0.1 μg/mL for calycosin, 0.5 μg/m L and 0.1 μg/m L formonetion, respectively. The standard recoveries were in the range of 96.5%-104.7%. The developed method has successfully been used for the determination of four main flavonoids in Astragali Radix from various sources and can be used for identification, differentiation and quality evaluation of Astragali Radix.

Analysis on Genetic Diversity of Radix Astragali by ISSR Markers

Radix Astragali has been an important traditional Chinese herbal medicine for over 2000 years. It is derived from two plant species, namely, Astragalus mongholicus [Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao] and Astragalus membranaceus [Astragalus membranaceus (Fisch.) Bge.] (Leguminosae ), according to the Pharmacopoeia of the People’s Republic of China. In this study, the genetic diversity and genetic relationships of Radix Astragali in China were analyzed by Inter-Simple Sequence Repeat (ISSR) markers. A total of 25 highly polymorphic ISSR primers were selected to amplify 95 Radix Astragali samples. Among 273 DNA bands amplified, 213 are polymorphic (percentage of polymorphic bands: 78%). The average value of the amplified bands was 10.9 for each primer, and the number varied from 4 to 20. The genetic diversity of the 95 Radix Astragali samples was analyzed by using POPGENE 1.32 software. The Nei’s genetic diversity index (h) and Shannon’s information index (I ) were 0.3590 and 0.5308, respectively, which indicated the abundant genetic diversity of Radix Astragali . The level of genetic diversity in A. membranaceus (h: 0.3109, I : 0.4657) was slightly lower than that in A. mongholicus (h: 0.3364, I : 0.4969). Considering the average genetic similarity coefficient by NTSYS analysis to cluster the A. membranaceus of nine habitats and A. mongholicus of five habitats, Radix Astragali samples were clustered into two groups according to place of origin. This clustering is different from traditional clustering, which divides groups according to species. Results obtained from this study will provide a theoretical basis for the molecular study on germplasm resources of Radix Astragali .

Network pharmacology research and experimental verification of Huangqi(Astragalus Radix)and Jinyingzi(Rosae Laevigatae Fructus)in treating benign prostatic hyperplasia

Objective This study aimed to analyze the mechanism of action of Huangqi(Astragalus Radix,HQ)-Jinyingzi(Rosae Laevigatae Fructus,JYZ)in the treatment of benign prostatic hyperplasia(BPH)based on network pharmacology and to verify the prediction through animal experimentation.Methods Based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN-TCM)databases,and literature,the active components and related target genes of HQ and JYZ were screened.The BPH target genes were screened based on the DisGeNET and GeneGards databases,and Excel was used to merge and remove duplicates.The Perl language was used to obtain drug-BPH target genes by intersecting shared target genes.A drug-component-target gene network diagram was constructed using Cytoscape software.The drug-BPH intersection target genes were inputted into the STRING database,and the key target genes were selected according to the degree algorithm.The output formed the basis for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses to determine the potential mechanism of HQ and JYZ in BPH treatment.High,medium,and low doses of HQ and JYZ extract were used to intervene in BPH rats,and then the prostate volume,wet weight,and prostate index of the BPH rats were determined.Changes in prostate histopathology and microvessel density(MVD)were evaluated using immunohistochemistry,and the optimal HQ and JYZ extract dose was confirmed.Finally,the optimal dose was used to intervene in a BPH rat model,and AKT1 and VEGF expressions were examined by immunohistochemistry.Results Based on network pharmacology,33 active components and 772 target genes were identified from HQ and JYZ,along with 817 BPH target genes and 112 drug-BPH common target genes.Among them were 10 key target genes,including AKT1,JUN,MAPK1,IL-6,TNF,ESR1,and VEGFA.KEGG enrichment analysis revealed 135 signaling pathways,including PI3K/AKT,IL-17,TNF,p53,MAPK,VEGF,JAK-STAT,and NF-κB pathways.The animal experiment showed that HQ and JYZ significantly improved prostate volume,wet weight,prostate index,and prostate histopathology of BPH rats,reducing MVD.In addition,HQ and JYZ inhibited the expression of AKT1 and VEGF in the prostate tissue of rats,promoted epithelial cell apoptosis,and inhibited angiogenesis,consistent with the prediction.Conclusion The combination of HQ and JYZ is effective for BPH therapy through multi-compound and multi-target collaboration.Its possible mechanism in treating BPH includes regulation of AKT1,VEGF protein,PI3K/Akt,and VEGF signaling pathways related to apoptosis,angiogenesis,and inflammation,with potential for clinical use and 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.

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.

Transcriptome Analysis of Gene in Mouse M-1 Cells Revealing the Functional Mechanism of Astragali Radix Extract

Astragali Radix (AR), the dried root of legumes, belongs to the Qi-invigorating herbs in traditional Chinese medicine and plays an important role in the treatment of many diseases. In order to understand the mechanism of action of AR extract. We used AR extract to treat M-1, mouse kidney cells, and used transcriptome sequencing technology to detect the genomic transcription level of the cells under the action of AR at different concentrations and times. The results showed that after a low concentration of AR treatments on the cells, the expression of genes related to cell growth and cellular immune response changed significantly, among which multiple genes are related to mitochondrial function, while high concentrations of AR affected the expression of histones and disease-related genes. It showed that the low concentration of AR extract can achieve the effect of invigorating Qi by regulating the function of mitochondria. In addition, several important genes and pathways were identified as potential targets of AR activation. The research not only clarified the main molecular biological mechanism of AR invigorating Qi, but also provided experimental basis and cellular physiology reference for the further clinical application of AR.

Therapeutic effects of Astragali Radix on diabetic foot:a clinical randomized controlled trial

Objective:This study was conducted to evaluate the efficacy of Astragalus on diabetic foot,as well as the effects on the levels of serum VEGF,bFGF,MMP-2,and inflammatory factors in patients,and to provide a scientific basis for the treatment of diabetic foot with the traditional Chinese medicine Astragalus.Methods:By taking 100 cases of diabetic foot patients who were admitted to the metabolic internal medicine division of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine and met the criteria of natriuresis during April 2021-April 2023 as the study subjects,and according to the random number method,all patients were divided into the control group and the observation group,with 50 cases in each group.In the control group,only basic treatment was carried out,while in the observation group,Astragalus injection was added based on the control group.After 8 weeks of treatment,the clinical efficacy,serum VEGF,bFGF,MMP-2,and inflammatory factor levels of the patients in the two groups were compared,respectively.Results:The total clinical efficiency of patients in the observation group was significantly better than that in the control group(χ^(2)=5.01,P<0.05).The inflammatory factor indexes decreased substantially in both groups.However,the decrease in the observation group was significantly higher than in the control group(P<0.05).Compared with the control group,serum VEGF and bFGF were considerably higher in the observation group,while MMP-2 was significantly lower(P<0.05).Conclusion:Astragali Radix is clinically effective in the diabetic foot,which can induce vascular endothelial repair and reduce the level of inflammatory factors,to improve the inflammatory state of patients and promote the restoration of ulcerated wound tissue,which is worth promoting in clinical practice.

A Preliminary Study in the Effect of <i>Astragali radix</i>, a Qi-Invigorating Herb, on Mitochondria: Insights at Cellular Level and Mouse Tissues Level

Objectives: To explore the effects of Qi-invigorating herbs on mitochondrial function using cultured cells and animal organs. Methods: Using water extracts of Astragali radix, we investigated the effect of “Qi-invigoration” on M-1 renal cells and mouse organs in-vitro including total adenylate production (TAP), reactive oxygen species (ROS) levels, and mitochondrial membrane potential (MMP). We also examined the effect on antioxidant capacity by conducting an analysis of superoxide dismutase (SOD) and glutathione (GSH). Results: 1) Astragali radix increased mitochondrial TAP generation and decreased ROS levels in both mouse kidney tissues and M-1 renal cells. 2) Astragali radix also significantly increased MMP and GSH levels in M-1 cells, but in the kidney tissue, there was no significant change in MMP levels and a decrease in GSH levels. 3) Astragali radix stimulated TAP levels in the heart, spleen, lung, kidney and skeletal muscle tissue, which was accompanied by the reduction of ROS. 4) For the meridian organs that Astragali radix belongs to, the energy production and antioxidant capacity were boosted simultaneously. Conclusions: These results provide new insights for the biochemical basis of Qi-invigoration and the meridian tropism theory for this Qi-invigorating herb.

Understanding the Use of Radix Astragali Seu Hedysari in the Treatment of Edema Due to Renal Diseases

Radix Astragali seu Hedysari is warm in nature and slightly sweet in taste.It belongs to the lung,spleen,and kidney meridians.It passes through a waterway,dredges the triple energizer,and has a significant effect on edema due to renal diseases.Ancient doctors believed that Radix Astragali seu Hedysari is excellent in tonifying the middle,supplementing Qi,and dredging through the triple energizer.In modem times,many doctors have found that whether it is used as a single drug or in combination with other drugs,it is widely used in clinical practice and has a good effect in inducing diuresis.

黄芪熊竹素对脂多糖诱导肺泡上皮细胞氧化应激损伤的改善作用

目的探讨黄芪熊竹素对脂多糖(LPS)诱导的肺泡上皮细胞A549氧化应激损伤的改善作用,以及对核因子-红系2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)信号通路的影响。方法实验设空白对照组(DMEM培养基)、模型组(DMEM培养基+LPS 200 ng/mL),地塞米松组(DMEM培养基+LPS 200 ng/mL+地塞米松200 ng/mL)及黄芪熊竹素低、高剂量组(DMEM培养基+LPS 200 ng/mL+黄芪熊竹素400、800 ng/mL)。于570 nm波长处检测吸光度(OD)以计算细胞存活率;流式细胞仪检测细胞凋亡水平;检测氧化应激因子活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平;RT-PCR法及Western blot法分别检测细胞Nrf2、HO-1 mRNA和蛋白表达水平。结果与模型组比较,各给药组细胞OD值,细胞存活率,单细胞克隆形成数目,SOD水平,Nrf2、HO-1 mRNA和蛋白表达水平均显著升高,细胞凋亡率,ROS、MDA水平均显著降低(P<0.05)。结论黄芪熊竹素对LPS诱导的A549细胞氧化应激损伤有显著改善作用,其机制可能与促进细胞Nrf2、HO-1的高表达有关。

黄芪免疫调节活性成分及其药理作用进展

免疫性疾病发生率逐年增长,目前免疫抑制剂、糖皮质激素、生物制剂等为其主要治疗手段,但存在治疗过度、治疗不足、毒副作用大等问题。黄芪及其有效成分免疫调节活性显著,具有双向调节免疫、抗炎、抗氧化应激、抗肿瘤等药理作用,在治疗肿瘤、重症肌无力、炎症性肠病、特应性皮炎、银屑病等免疫性疾病方面疗效突出。黄芪可多途径、多靶点防治免疫性疾病,现综述黄芪及其活性成分(黄芪皂苷Ⅲ、黄芪皂苷Ⅳ、环黄芪醇及黄酮类化合物)调节免疫的作用及机制,为后续临床研究提供依据。

基于TGF-β/Smad信号通路探讨黄芪有效成分治疗纤维化疾病综述

黄芪是一种传承千年的传统中药材。现代药物研究已证实黄芪组成成分丰富,比如皂苷类和多糖类成分,其中黄芪甲苷和黄芪多糖都是其产生药理作用的主要活性成分。现代医学研究表明,黄芪能改善肺纤维化进程、对抗肾间质纤维化、降低肝脏和心肌纤维化疾病程度。通过对黄芪的基于信号通路TGF-β/Smad抗纤维化功能进行综述,深入探究其主要成分作用特点,进一步丰富其药理活性机制,为中西医临床提供药效理论依据。

黄芪-葛根配伍对T2DM胰岛素抵抗大鼠铁死亡的影响及机制研究

目的探讨黄芪-葛根配伍对2型糖尿病(T2DM)胰岛素抵抗(IR)大鼠肝细胞铁死亡的影响及潜在机制。方法将雄性SD大鼠60只分为对照组(12只)与造模组(48只),造模组大鼠以高脂饲料连续喂养4周后按25 mg/kg的剂量一次性尾静脉注射1%链脲佐菌素溶液以复制T2DM IR大鼠模型。将造模成功的大鼠随机分为模型组、黄芪-葛根配伍组[QG组,4.05 g/(kg·d)灌胃]、铁死亡抑制剂ferrostatin-1组(Fer-1组,5 mg/kg隔日1次腹腔注射)、黄芪-葛根配伍+铁死亡诱导剂erastin组[QG+erastin组,4.05 g/(kg·d)灌胃+erastin 10 mg/(kg·d)腹腔注射]。干预4周后,检测各组大鼠血清空腹血糖(FBG)、空腹胰岛素(FINS)水平,并计算稳态模型胰岛素抵抗指数(HOMA-IR)和胰岛素敏感指数的自然对数(IAI);检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)水平,血清Fe^(2+)及Fe水平,肝脏组织中谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平以及烟酰胺腺嘌呤二核苷酸磷酸(NADP+)/还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)和活性氧(ROS)水平;观察其肝脏组织病理形态,检测肝脏组织谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链1(FTH1)、长链脂酰辅酶A合成酶3(ACSL3)、ACSL4、线粒体铁蛋白(FTMT)、胱氨酸/谷氨酸反向转运蛋白(xCT)的蛋白表达情况。结果与对照组比较,模型组大鼠肝脏组织内肝细胞排列紊乱、肿胀,细胞核染色加深,可见较多炎症细胞浸润,大量肝细胞空泡及脂肪变性;FBG(给药后)、TC、TG、LDL-C、AST、ALT、FINS、MDA、ROS、Fe^(2+)、Fe水平及HOMA-IR,NADP+/NADPH和ACSL4蛋白的表达均显著升高或上调(P<0.01);HDL-C、GSH、SOD水平,IAI及GPX4、FTH1、ACSL3、FTMT、xCT蛋白的表达均显著降低或下调(P<0.01)。与模型组比较,QG组和Fer-1组大鼠肝脏组织病理损伤及上述指标水平均有不同程度改善,且大部分指标的变化差异有统计学意义(P<0.01或P<0.05)。与QG组比较,QG+erastin组大鼠上述指标水平的改善均被显著逆转(P<0.01)。结论黄芪-葛根配伍水煎液可降低T2DM IR大鼠FBG水平,减轻其IR程度和肝脏组织铁负荷,缓解其肝脏组织病理损伤,上述作用与其抑制铁死亡有关。

基于网络药理学和实验验证研究黄芪-党参药对干预动脉粥样硬化的作用机制

基于网络药理学、分子对接及实验验证探究中药药对黄芪-党参(Astragali Radix-Codonopsis Radix,AR-CR)治疗动脉粥样硬化(atherosclerosis,AS)的分子作用机制。首先通过检索TCMSP、PubChem、SwissTargetPrediction、UniProt、GeneCards等数据库,获取中药药对AR-CR的活性成分,预测该药对的作用靶点,筛选动脉粥样硬化的相关靶点基因,然后利用Venny平台、STRING数据库、Cytoscape(Version 3.8.2)软件进行拓扑分析获取AR-CR治疗动脉粥样硬化的关键靶点,使用DAVID数据库对所获得的关键靶点进行GO和KEGG富集分析,并借助Auto Dock tools、Auto Dock cina对核心蛋白与活性成分完成分子对接,最后,利用氧化型低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导人脐静脉内皮细胞,建立AS细胞模型进行体外生物学验证。AR-CR共检索到34个活性化合物,并预测潜在化合物靶点426个,通过与875个AS靶点进行交际映射,获得AR-CR治疗AS关键靶点69个,筛选出3,9-二邻甲基苯胺、7-甲氧基-2-甲基异黄酮、5α-豆甾烷-3,6-二酮、木犀草素4个主要活性化合物,丝氨酸/苏氨酸蛋白激酶(serine/threonine-protein kinase,AKT1)、肿瘤蛋白p53(tumor protein p53,TP53)、丝裂原激活蛋白激酶3(mitogen-activated protein kinase 3,MAPK3)等25个核心靶点。KEGG通路富集分析得到关键通路为糖基化终末产物(advanced glycation end-product,AGE)/糖基化终末产物受体(receptor for advanced glycation endproduct,RAGE)信号通路和脂质与动脉粥样硬化信号通路等。分子对接结果显示4个主要活性化合物与核心靶点蛋白均能连接,其中与TP53的结合活性最强。体外实验结果表明低、中、高剂量的AR-CR能够促进动脉粥样硬化模型细胞增殖,抑制其凋亡,并促进TP53 mRNA及TP53蛋白表达。综上,该研究初步揭示AR-CR治疗AS的作用机制,与网络药理学及分子对接预测的TP53因子相关,通过调控TP53因子促进AS模型细胞增殖,抑制其凋亡,为临床治疗AS提供了理论基础。

黄芪抗病毒药效物质基础和作用机制研究进展

黄芪是扶正补气的中药,是临床常用的中药之一。黄芪含有多种化学成分,包括多糖类、黄酮类和皂苷类等,具有抗病毒、抗肿瘤、调节免疫功能等药理作用,其中抗病毒作用突出。黄芪抗病毒可通过多种方式实现,如直接杀灭病毒、阻滞病毒对细胞的吸附穿入、抑制病毒的复制和释放、抑制病毒RNA和蛋白质的合成与表达,还可促进免疫器官的发育、增强先天免疫、增强细胞免疫和体液免疫及调节相关信号通路等。黄芪抗病毒具有安全、低毒、高效等特点,值得进一步开发研究。总结了黄芪抗病毒的药效物质基础及作用机制,为其开发利用及其中药临床研究提供理论依据。