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astragalosideⅣameliorates insulin induced insulin resistance in HepG2 cells through reactive oxygen species mediated c-Jun N-terminal kinase pathway

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Journal of Traditional Chinese Medicine 2023年第1期43卷 60-67页

作者： YE Xiaomei XING Xiaowei YUAN Kangrui WANG Dongming WU Dudu CHEN Zhi YU Zhiqiang School of Pharmacy Guangdong Medical UniversityDongguan 523808China Department of Clinical Pharmacy SSL Central Hospital of Dongguan City(the Third People's Hospital of Dongguan City)Dongguan 523326China Department of Pharmacy Central People's Hospital of ZhanjiangGuangzhou 510515China School of Pharmaceutical Sciences Guangdong Provincial Key Laboratory of New Drug ScreeningSouthern Medical UniversityZhanjiang 524037China

OBJECTIVE:To investigate the effects and elucidate the mechanism of astragaloside IV(AS-IV)for insulin resistance(IR)and type 2 diabet es mellitus(T2DM).METHODS:CCK8 kit was used to detect cell viability,glucose detection kit was used to detect the concentration of glucose in cell supernatant,reactive oxygen species(ROS)detection kit and Western blot were used to explore the mechanism of astragaloside IV(AS-IV)in improving IR.A diabetic rat model was also established by feeding high sugar and fat diet and streptozotocin(STZ)injection.After treatment with AS-IV,rosiglitazone(ROZ),or normal saline,the fasting blood glucose(FBG),C peptide(C-P),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and the glucose tolerance were assessed.RESULTS:AS-IV could effectively reduce the content of ROS and increase the glucose uptake in high insulintreated IR-type HepG 2 cells.The results of molecular mechanisms indicated that AS-IV could improve insulin resistance by reducing JNK phosphorylation and regulating c-Jun N-terminal kinase(JNK)downstream protein expression.Additionally,AS-IV could significantly reduce the levels of FBG,TNF-α,IL-6 and the glucose tolerance in diabetic rats(P<0.05 or<0.01).The high and medium dose groups of AS-IV could significantly increase the C-P levels in diabetic rats(P<0.05 or<0.01).CONCLUSIONS:Our results indicated that AS-IV improve liver IR through the JNK pathway and ROS,which meant a new molecular target for the treatment of diabetes.The AS-IV also helped to prevent and improved the insulin resistance of rats.

关键词： astragalosideⅣ Insulin resistance Glucose consumption ROS JNK HepG2 cells

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astragalosideⅣplays a role in reducing radiation-induced liver inflammation in mice by inhibiting thioredoxin-interacting protein/nod-like receptor protein 3 signaling pathway

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Journal of Traditional Chinese Medicine 2023年第1期43卷 87-94页

作者： DING Yanping DONG Xiaoqing MA Yifan CHEN Lili ZHOU Jie LI Xinyan SHAO Baoping College of Life Science Northwest Normal UniversityLanzhou 730000China College of Life Science Lanzhou UniversityLanzhou 730000China

OBJECTIVE:To investigate the efficacy of astragaloside IV(AS-IV)on radiation-induced liver inflammation in mice.METHODS:The mice were divided into normal group,dimethyl sulfoxide solvent group,irradiation group(IR),irradiation+AS-IV(20 mg/kg)group(IR+AS-20)and irradiation+AS-IV(40 mg/kg)group(IR+AS-40).One month after intraperitoneal injection of AS-IV,the mice were irradiated with 8Gry Co60γ,the blood was collected for biochemical analysis,and the liver was collected for hematoxylin-eosin staining,immunofluorescence and electron microscopic observation,oxidative stress,and Western blot analysis.RESULTS:The AS-IV treatment significantly ameliorated the pathological morphology of liver and reduced the alanine aminotransferase and aspertate aminotransferase levels in serum induced by radiation;AS-IV treatment also significantly reduced the expression of inflammatory factors tumor necrosis factor alpha and interleukin 6 and antagonized malonaldehyde content and superoxide dismutase activity in liver caused by radiation;in addition,AS-IV treatment can significantly inhibited the positive expression of thioredoxin-interacting protein(TXNIP)and nod-like receptor protein 3(NLRP3)inflammasome in liver tissue after radiation;The expression of TXNIP,NLRP3 inflammasome,apoptosisassociated speck-like protein containing a CARD,cysteinyl aspartate-specific proteinase 1 and interleukin 1beta in the AS-IV prevention group decreased significantly compared to the radiation group.CONCLUSIONS:These findings suggested that Co60γradiation can cause structural and functional damage to the liver,which may be related to the NLRP3 mediated inflammatory pathway;AS-IV may play a protective role by inhibiting the TXNIP/NLRP3 inflammasome signaling pathway in the radiation-induced liver injury model.

关键词： radiation,ionizing astragalosideⅣ liver inflammation NLR family,pyrin domain-containing 3 protein thioredoxin-interacting protein

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Effect of astragaloside Ⅳ on the immunoregulatory function of adipose-derived mesenchymal stem cells from patients with psoriasis vulgaris

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Journal of Traditional Chinese Medicine 2022年第4期42卷 513-519页

作者： YIN Xiuping ZHANG Xiaotong ZHU Rongjia SONG Ping Department of Dermatology Guang'anmen HospitalChina Academy of Chinese Medical SciencesBeijing 100053China Beijing Key Laboratory of New Drug Development and Clinical Trial of Stem Cell Therapy(BZ0381)/Center of Excellence in Tissue Engineering Institute of Basic Medical Sciences CAMSSchool of Basic Medicine PUMCBeijing 100005China

OBJECTⅣE:To compare the phenotype and adipogenic and osteogenic differentiation capacities of adiposederived mesenchymal stem cells(AMSCs)isolated from patients with psoriasis vulgaris and healthy donors,and to explore the effects of astragalosideⅣ,a Traditional Chinese Medicine,on the immunoregulatory function of AMSCs.METHODS:AMSCs were isolated from human adipose tissue and cultured for three generations in vitro.Cell phenotype and cell cycle analysis were performed by flow cytometry.Adipogenic and osteogenic differentiation of AMSCs was examined by lipid(oil red O)and alkaline phosphatase staining,respectively.Expression of inflammatory mediators was examined by real-time quantitative polymerase chain reaction analysis,and proliferation was quantified using the cell counting kit-8 assay.RESULTS:Expression of CD29,CD44,and CD73 was higher in AMSCs from healthy donors than psoriasis patients,while the reverse was true for expression of CD45,CD31,and HLA-DR.AMSCs from psoriasis patients had a greater ability to undergo adipogenic differentiation than cells from healthy donors,whereas there was no significant difference in osteogenic differentiation between AMSCs from the two sources.Compared with AMSCs from healthy donors,psoriasis patient-derived AMSCs expressed lower levels of the anti-inflammatory cytokines interleukin-10 and transforming growth factor-β(TGF-β)and the immune checkpoint ligand programmed cell death 1 ligand 1(PDL1)(PastragalosideⅣhad no significant effect on proliferation but increased the expression of TGF-βand PDL1 and decreased the expression of IFN-γand TNF-α.CONCLUSION:AMSCs from patients with psoriasis vulgaris display abnormal proliferation and adipogenesis and an enhanced pro-inflammatory phenotype.These defects were normalized by treatment with astragalosideⅣ,suggesting that this Traditional Chinese Medicine may be useful for

关键词： psoriasis mesenchymal stem cells cell proliferation astragalosideⅣ adipogenesis osteogenesis immune regulation

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astragaloside Ⅳ suppresses post-ischemic natural killer cell infiltration and activation in the brain:involvement of histone deacetylase inhibition

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Frontiers of Medicine 2021年第1期15卷 79-90页

作者： Baokai Dou Shichun Li Luyao Wei Lixin Wang Shiguo Zhu Zhengtao Wang Zunji Ke Kaixian Chen Zhifei Wang School of Basic Medical Sciences Shanghai University of Traditional Chinese MedicineShanghai 201203China Institute of Chinese Materia Medica Shanghai University of Traditional Chinese MedicineShanghai 201203China Academy of Integrative Medicine Shanghai University of Traditional Chinese MedicineShanghai 201203China

Natural killer(NK)cells,a type of cytotoxic lymphocytes,can infiltrate into ischemic brain and exacerbate neuronal cell death.astragaloside Ⅳ(ASIV)is the major bioactive ingredient of Astragalus membranaceus,a Chinese herbal medicine,and possesses potent immunomodulatory and neuroprotective properties.This study investigated the effects of ASIV on post-ischemic brain infiltration and activation of NK cells.ASIV reduced brain infarction and alleviated functional deficits in MCAO rats,and these beneficial effects persisted for at least 7 days.Abundant NK cells infiltrated into the ischemic hemisphere on day 1 after brain ischemia,and this infiltration was suppressed by ASIV.Strikingly,ASIV reversed NK cell deficiency in the spleen and blood after brain ischemia.ASIV inhibited astrocyte-derived CCL2 upregulation and reduced CCR2+NK cell levels in the ischemic brain.Meanwhile,ASIV attenuated NK cell activating receptor NKG2D levels and reduced interferon-γproduction.ASIV restored acetylation of histone H3 and the p65 subunit of nuclear factor-κB in the ischemic brain,suggesting inhibition of histone deacetylase(HDAC).Simultaneously,ASIV prevented p65 nuclear translocation.The effects of ASIV on reducing CCL2 production,restoring acetylated p65 levels and preventing p65 nuclear translocation were mimicked by valproate,an HDAC inhibitor,in astrocytes subjected to oxygen-glucose deprivation.Our findings suggest that ASIV inhibits post-ischemic NK cell brain infiltration and activation and reverses NK cell deficiency in the periphery,which together contribute to the beneficial effects of ASIV against brain ischemia.Furthermore,ASIV’s effects on suppressing NK cell brain infiltration and activation may involve HDAC inhibition.

关键词： astragalosideⅣ brain ischemia natural killer cells histone deacetylase nuclear factor-κB

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黄芪甲苷拮抗Aβ1-42所致大鼠氧化应激,神经炎症和认知功能损伤（英文）

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Chinese Medical Sciences Journal 2018年第1期33卷 29-37页

作者：潘艳芳贾晓涛宋二飞彭小忠陕西中医药大学病理教研室西安交通大学附属西安市中心医院神经内科加拿大约克大学生物学系中国医学科学院基础医学研究所生物化学与分子生物学系医学分子生物学国家重点实验室

To investigate the neuroprotective action of astragalosideⅣ(AS-Ⅳ)on spatial learning and memory impairment induced by amyloid-beta 1-42(Aβ1-42)in rats and elucidate its underlying molecular mechanisms.Methods Adult-male Sprague-Dawley rats(230-250 g)were divided into six groups randomly:control,Aβ1-42,AS-Ⅳ,Aβ1-42 plus 5 mg/kg·d AS-Ⅳ,Aβ1-42 plus 25 mg/kg·d AS-Ⅳ,and Aβ1-42 plus 50 mg/kg·d AS-Ⅳgroups.Aβ1-42 were delivered by intracerebroventricular injection under the guidance of a brain stereotaxic apparatus.The Morris water maze test(hidden platform test,probe trials,visible platform test)was performed one week after Aβ1-42 injection to obtain the ability of rat spatial learning and memory.AS-Ⅳ(5,25 and 50 mg/kg·d)was administrated intraperitoneally once per day from the 8th day after Aβ1-42 injection for 5 consecutive days.Average escape latencies,distances for searching for the platform under water and the percentage of total time elapsed and distance swam in the right quadrant after removing platform were determined by behavior software system.

关键词： amyloid-βprotein astragalosideⅣ spatial learning and memory oxidative stress neuroinflammation

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Effect of astragaloside Ⅳ on lipid and glucose metabolism in acute myocardial infarction through PPARγ pathway

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中国药理学与毒理学杂志 2019年第9期33卷 680-681页

作者： ZHANG Qian WANG Xiao-ping WANG Yong LI Chun WANG Wei Beijing University of Chinese Medicine Beijing 100029China

OBJECTIVE To investigate the effects of astragaloside IV(which can be extracted from the traditional Chinese medicine Astragalus membranaceus)on lipid and glucose metabolism in acute myocardial infarction(AMI).METHODS Model of heart failure(HF)after AMI was established with ligation of left anterior descending artery on Sprague-Dawley(SD)rats.The rats were divided into three groups:sham,model and astragaloside IV treatment group.Twenty-eight days after treatment(astragaloside IV,20 mg·kg-1 daily),hematoxylin-eosin(HE)staining was applied to visualize cardiomyocyte morphological changes.High performance liquid chromatography(HPLC)was performed to assess the contents of adenosine phosphates in heart.Positron emission tomography and computed tomography(PET-CT)was conducted to evaluate the cardiac glucose metabolism.Expressions of key molecules such as peroxisome proliferatoractivated receptor γ(PPARγ),sterol carrier protein 2(SCP2)and long chain acyl CoA dehydrogenase(ACADL)were measured by Western blotting and immunohistochemistry.Oxygen-glucose deprivation-reperfusion(OGD/R)-induced H9C2 injury cardiomyocyte model was adopted for potential mechanism research in vitro.RESULTS Treatment with astragaloside Ⅳ rescued hearts from structural and functional damages as well as inflammatory infiltration.Levels of adenosine triphosphate(ATP)and energy charge(EC)in astragaloside IV group were also up-regulated compared to model group.Further results demonstrated that critical enzymes both in lipid metabolism and glucose metabolism compro mised in model group compared to sham group.Intriguingly,astragalosideⅣcould up-regulate critical enzymes including ACADL and SCP2 in lipid metabolism accompanying with promoting effect on molecules in glycolysis simultaneously.Results on upstreaming signaling pathway demonstrated that astragaloside Ⅳ could dramatically increase the expres sions of PPARγ.In vitro study suggested the efficacy of astragalosideⅣcould be blocked by T0070907,a selective PPARγ inhibitor.CONCLUSION astragaloside IV

关键词： acute myocardial infarction astragalosideⅣ lipid and glucose metabolism PPARγpathway

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Review of the pharmacological effects of astragalosideⅣand its autophagic mechanism in association with inflammation

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World Journal of Clinical Cases 2022年第28期10卷 10004-10016页

作者： Ying Yang Meng Hong Wen-Wen Lian Zhi Chen State Key Laboratory for Diagnosis and Treatment of Infectious Diseases National Clinical Research Center for Infectious DiseasesNational Medical Center for Infectious DiseasesCollaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesThe First Affiliated HospitalZhejiang University School of MedicineHangzhou 310003Zhejiang ProvinceChina

Astragalus membranaceus Bunge,known as Huangqi,has been used to treat various diseases for a long time.astragalosideⅣ(AS-Ⅳ)is one of the primary active ingredients of the aqueous Huangqi extract.Many experimental models have shown that AS-Ⅳexerts broad beneficial effects on cardiovascular disease,nervous system diseases,lung disease,diabetes,organ injury,kidney disease,and gynaecological diseases.This review demonstrates and summarizes the structure,solubility,pharmacokinetics,toxicity,pharmacological effects,and autophagic mechanism of AS-Ⅳ.The autophagic effects are associated with multiple signalling pathways in experimental models,including the PI3KI/Akt/m TOR,PI3KⅢ/Beclin-1/Bcl-2,PI3K/Akt,AMPK/m TOR,PI3K/Akt/m TOR,SIRT1–NF-κB,PI3K/AKT/AS160,and TGF-β/Smad signalling pathways.Based on this evidence,AS-Ⅳcould be used as a replacement therapy for treating the multiple diseases referenced above.

关键词： astragalosideⅣ Pharmacological effect Autophagy Inflammation

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astragalosideⅣPrevents the Mitochondrial Permeability Transition Pore Opening by Inhibiting Glycogen Synthase Kinase 3β via the NO/cGMP/PKG Pathway in Cardiac Cells

AstragalosideⅣPrevents the Mitochondrial Permeability Trans...

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中国生理学会心血管生理学术研讨会

作者： Yonggui He~(1,2),Yuxi Xie~2,Huan Zheng~2,Yidong Zhang~2,Jing Guo~2,Jinkun Xi~(2*) 1.Department of Internal Medicine 2.Heart Institute,Hebei United University Tangshan 063000

Aim:astragalosideⅣ,a major ingredient of traditional Chinese herb Radix Astragali,has been widely used for treatments of cardiovascular diseases in China,but the exact cellular and molecular mechanisms underlying its action remain unclear.We aimed to test if astragalosideⅣmodulates mitochondrial permeability transition pore(mPTP) opening through glycogen synthase kinase-3β(GSK-3β) in H9c2 cardiac cells.Further more,the molecular mechanism was studied,by which astragaloside IV inhibits GSK-3βactivity in this study.Methods:Rat heart tissue-derived H9c2 cells were purchased from ATCC.And cardiac H9c2 cells were exposed to HO(600μM) for 20 min to induce mitochondrial oxidant damage.Fluorescence dyes including tetramethylrhodamine ethyl ester(TMRE,red) and 4-amino-5-methylamino-2’, 7’-difluorofluorescein(DAF-FM,green) diacetate were used to image mitochondrial membrane potential(Δψm) and nitric oxide(NO),respectively.Fluorescence images were obtained with laser-scanning confocal microscopy.The red fluorescence was excited with a 543 nm line of He-Ne laser line and imaged through a 560 nm long-path filter.The green fluorescence was excited at 488 nm and imaged through a 525 nm long-path filter.The cell viability was assessed by propidium iodide fluorometry using a fluorescence reader.Cells in 12-well plates coated with laminin were incubated in standard Tyrode solution,and fluorescence intensity was measured at the excitation and emission wavelengths of 540 and 590 nm,respectively.H9c2 cells were exposed to astragalosideⅣfor 20 min,the GSK-3β(Ser),Akt(Ser),and VASP(Ser) activities were determined by measuring their phosphorylation statuses with Western blot. Results:Cardiac cells treated with different dose of astragalosideⅣ(30μM-80μM),and 50μM astragalosideⅣsignificantly increased GSK-3βphosphorylation at Ser(121.1±2.7%) and prevented oxidative stress-induced loss of mitochondrial membrane potential(MMP) in H9c2 cardiac cells(89.7±4.3%vs.49.7±6.3%in control).The GSK-3βinhibitor SB216763(3μM) and the specific mPTP clos

关键词： astragalosideⅣ mPTP GSK-3β cGMP/PKG NO PI3K/Akt

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Spatial-resolved metabolomics reveals tissue-specific metabolic reprogramming in diabetic nephropathy by using mass spectrometry imaging

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Acta Pharmaceutica Sinica B 2021年第11期11卷 3665-3677页

作者： Zhonghua Wang Wenqing Fu Meiling Huo Bingshu He Yaqi Liu Lu Tian Wanfang Li Zhi Zhou Baili Wang Jianzhen Xia Yanhua Chen Jinfeng Wei Zeper Abliz Key Laboratory of Mass Spectrometry Imaging and Metabolomics(Minzu University of China) National Ethnic Affairs CommissionBeijing 100081China College of Life and Environmental Sciences Minzu University of ChinaBeijing 100081China New Drug Safety Evaluation Center Institute of Materia MedicaPeking Union Medical CollegeBeijing 100050China School Hospital Minzu University of ChinaBeijing 100081China Key Laboratory of Ethnomedicine of Ministry of Education School of PharmacyMinzu University of ChinaBeijing 100081China

Detailed knowledge on tissue-specific metabolic reprogramming in diabetic nephropathy(DN)is vital for more accurate understanding the molecular pathological signature and developing novel therapeutic strategies.In the present study,a spatial-resolved metabolomics approach based on air flowassisted desorption electrospray ionization(AFADESI)and matrix-assisted laser desorption ionization(MALDI)integrated mass spectrometry imaging(MSI)was proposed to investigate tissue-specific metabolic alterations in the kidneys of high-fat diet-fed and streptozotocin(STZ)-treated DN rats and the therapeutic effect of astragalosideⅣ,a potential anti-diabetic drug,against DN.As a result,a wide range of functional metabolites including sugars,amino acids,nucleotides and their derivatives,fatty acids,phospholipids,sphingolipids,glycerides,carnitine and its derivatives,vitamins,peptides,and metal ions associated with DN were identified and their unique distribution patterns in the rat kidney were visualized with high chemical specificity and high spatial resolution.These region-specific metabolic disturbances were ameliorated by repeated oral administration of astragaloside Ⅳ(100 mg/kg)for 12 weeks.This study provided more comprehensive and detailed information about the tissue-specific metabolic reprogramming and molecular pathological signature in the kidney of diabetic rats.These findings highlighted the promising potential of AFADESI and MALDI integrated MSI based metabolomics approach for application in metabolic kidney diseases.

关键词： Spatial-resolved metabolomics Mass spectrometry imaging Diabetic nephropathy Metabolic reprogramming astragalosideⅣ

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