Functional characterization of a cycloartenol synthase and four glycosyltransferases in the biosynthesis of cycloastragenol-type astragalosides from Astragalus membranaceus

Astragalosides are the main active constituents of traditional Chinese medicine Huang-Qi,of which cycloastragenol-type glycosides are the most typical and major bioactive compounds.This kind of compounds exhibit various biological functions including cardiovascular protective,neuroprotective,etc.Owing to the limitations of natural sources and the difficulties encountered in chemical synthesis,re-engineering of biosynthetic machinery will offer an alternative and promising approach to producing astragalosides.However,the biosynthetic pathway for astragalosides remains elusive due to their complex structures and numerous reaction types and steps.Herein,guided by transcriptome and phylogenetic analyses,a cycloartenol synthase and four glycosyltransferases catalyzing the committed steps in the biosynthesis of such bioactive astragalosides were functionally characterized from Astragalus membranaceus.AmCAS1,the first reported cycloartenol synthase from Astragalus genus,is capable of catalyzing the formation of cycloartenol;AmUGT15,AmUGT14,AmUGT13,and AmUGT7 are four glycosyltransferases biochemically characterized to catalyze 3-O-xylosylation,3-O-glucosylation,25-O-glucosylation/O-xylosylation and 2’-O-glucosylation of cycloastragenol glycosides,respectively.These findings not only clarified the crucial enzymes for the biosynthesis and the molecular basis for the structural diversity of astragalosides in Astragalus plants,also paved the way for further completely deciphering the biosynthetic pathway and constructing an artificial pathway for their efficient production.

Transformation of astragalosides from Radix Astragali under acidic,neutral,and alkaline extraction conditions monitored by LC-ESI-TOF/MS

AIM: To reveal the profile of astragalosides for better quality evaluation of Radix Astragali, this study was aimed to investigate the transformation of astragalosides under different conditions. METHOD: Seven major astragalosides were selected for evaluation under acidic, neutral and alkaline conditions. The transformation in real plant samples was also examined and the products were characterized by LC-ESI-TOF/MS. RESULTS: In weak acidic solution, all of the astragalosides are stable. In addition, the transformation ratios of the astragalosides under neutral and alkaline conditions were also obtained. CONCLUSION: In neutral solution, malonylastragaloside I was transformed to astragaloside I; and in alkaline solution, substituent group(s) in the xylose moiety of all the astragalosides were eliminated. Since astragalosdie IV is the basic skeleton structure of the astrgalosides, it is a common transformation product of other astragalosides.

Astragaloside IV Ameliorates Inflammatory Damage in Mice with Acute Liver Failure

Acute liver failure is a life-threatening clinical syndrome with a high mortality rate. Currently, the research on Astragaloside IV in liver diseases primarily focuses on liver cancer, and there is limited understanding of its mechanism in acute liver failure’s innate immunity. Therefore, this study aims to investigate the potential protective effect of Astragaloside IV on acute liver failure and its impact on innate immune cells. The study employed D-GalN/LPS-induced acute liver failure mouse models and employed various techniques such as a range of molecular and analytical techniques. The experimental results demonstrated that treatment with Astragaloside IV significantly reduced the inflammatory response, alleviated liver injury, and improved the survival rate of mice with acute liver failure induced by D-GalN/LPS. Further investigations revealed that AS-IV played a beneficial role by regulating the proportion of CD11b+Ly6Chi monocytes and the secretion of inflammatory cytokines and anti-inflammatory metabolites. These findings suggest that the pharmacological mechanism of AS-IV may involve targeted regulation of CD11b+Ly6Chi monocytes in both peripheral blood and liver. The implications of this study’s results are twofold. Firstly, they provide a basis for the clinical application of AS-IV in treating liver failure, offering potential therapeutic benefits. Secondly, they serve as a reference for further development of safer and more effective modified compounds.

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）

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.

黄芪甲苷孵育的脂肪源性干细胞对顺铂诱导的急性肾损伤小鼠的保护作用

目的:建立顺铂诱导的急性肾损伤(AKI)小鼠模型,尾静脉输注人脂肪源性干细胞(hADSC)及黄芪甲苷孵育的人脂肪源性干细胞(Ast-hADSC),探讨其对AKI小鼠肾损伤的修复作用。方法:应用PKH26标记hADSC及Ast-hADSC;雄性BALB/C裸鼠随机分为4组(normal组、model组、hADSC及Ast-hADSC干预组),干预组及模型组腹腔注射顺铂建立AKI模型,于建模后24 h尾静脉输注100μl hADSC及Ast-hADSC悬液,模型组给予等量生理盐水。分别留取建模后24 h及静脉输注后1 d、3 d、7 d、14 d小鼠血清及肾组织标本,检测血Scr、BUN水平,HE染色观察肾组织病理变化,TUNEL法检测肾小管上皮细胞凋亡,激光共聚焦观察hADSC及Ast-hADSC在小鼠肾组织的分布。结果:模型组小鼠Scr、BUN、肾组织病理改变及细胞凋亡区域均显著高于正常组及细胞干预组,其中Scr、BUN分别在第3天、第7天达到最高值,肾小管损伤严重;相同时间点Ast-hADSC组肾组织病理改变及细胞凋亡区域较hADSC组明显降低,至14 d可基本恢复至正常水平;激光共聚焦显示PKH26标记的hADSC及Ast-hADSC于14 d在小鼠肾组织有少量表达,其中少量红色荧光与肾小管上皮细胞呈融合状态。结论:hADSC移植可减轻AKI小鼠肾脏结构和功能的损伤;经Ast孵育后,可在一定程度上提高hADSC对受损肾组织的修复作用。

基于综合评分的中药复方“鼻敏康”提取工艺研究

目的：优选中药复方“鼻敏康”的最佳提取工艺并建立黄芪甲苷的HPLC-ELSD含量测定方法。方法：采用加热回流法，乙醇为提取溶剂，以浸膏率及黄芪甲苷含量为指标，对乙醇浓度、提取时间、液料比和提取次数进行单因素考查，优选出对浸膏率及黄芪甲苷含量较大影响的3个因素，按照L9（34）安排正交试验优选复方制剂鼻敏康的提取工艺。黄芪甲苷含量测定采用Diamonstil C18（250 mm＆#215;4.6 mm，5μm）色谱柱，流动相为水（A）-乙腈（B）,洗脱梯度：0-45 min，22%B；45-60 min，22%-32%B，流速1.0 mL＆#183;min-1，漂移管温度100℃，载气为N2，载气流速2.5 L＆#183;min-1，柱温：30℃。结果：复方制剂鼻敏康最佳提取工艺为8倍量70%乙醇,每次1.5 h，提取3次，结果表明乙醇浓度和提取次数存在显著性影响，最终确定提取工艺为8倍量70%乙醇,每次加热回流1.5 h，提取2次；黄芪甲苷含量测定线性范围为0.87-8.72μg，回归方程 Y=1.5454X＋5.8759，r =0.9997，平均收率95.05%，RSD值为2.64%。结论：优选的提取工艺稳定合理可行，适用于工业生产。

Astragaloside 对实验性自身免疫性脑脊髓炎小鼠的防治作用研究

目的:探讨Astragaloside对实验性自身免疫性脑脊髓炎(EAE)小鼠的保护作用及机制。方法:采用髓鞘MOG 35-55诱导C57BL/6雌性小鼠建立EAE模型,随机分为EAE对照组、Astragaloside高、低剂量组。造模的第3天起,采用腹腔注射给药,持续25 d。EAE对照组给予PBS;Astragaloside高、低剂量组分别给予Astragaloside溶液30 mg/(kg·d),15 mg/(kg·d);各组小鼠给药0.2 ml/(次·只),每天记录小鼠症状、体征、体重变化和临床评分。HE染色检测脊髓炎细胞浸润。ELISA法检测外周血中IL-1β、IL-17、TNF-α、IL-10的表达量。Western blot法检测脊髓组织中ROCKⅡ、P-MYPT1、p-NF-κB/p65的表达变化。结果:与EAE对照组比较,Astragaloside高、低剂量组均可明显降低平均最高临床评分,减轻EAE临床症状(P<0.01,P<0.05),高剂量组治疗效果优于低剂量组;Astragaloside可抑制中枢神经系统炎症细胞浸润,显著降低血清中IL-1β、IL-17的表达(P<0.05,P<0.001),增加血清IL-10水平(P<0.05),明显抑制ROCKⅡ、P-MYPT1、p-NF-κB/p65的表达(P<0.05)。结论:Astragaloside可通过抑制Rock通路和调节细胞因子的表达改善EAE的临床症状。

Astragaloside Ⅳ inhibits pathological functions of gastric cancer-associated fibroblasts

AIM To investigate the inhibitory effect of astragaloside IV on the pathological functions of cancer-associated fibroblasts,and to explore the underlying mechanism.METHODS Paired gastric normal fibroblast(GNF) and gastric cancer-associated fibroblast(GCAF) cultures were established from resected tissues. GCAFs were treated with vehicle control or different concentrations of astragaloside Ⅳ. Conditioned media were prepared from GNFs,GCAFs,control-treated GCAFs,and astragaloside Ⅳ-treated GCAFs,and used to culture BGC-823 human gastric cancer cells. Proliferation,migration and invasion capacities of BGC-823 cells were determined by MTT,wound healing,and Transwell invasion assays,respectively. The action mechanism of astragaloside Ⅳ was investigated by detecting the expression of micro RNAs and the expression and secretion of the oncogenic factor,macrophage colonystimulating factor(M-CSF),and the tumor suppressive factor,tissue inhibitor of metalloproteinase 2(TIMP2),in different groups of GCAFs. The expression of the oncogenic pluripotency factors SOX2 and NANOG in BGC-823 cells cultured with different conditioned media was also examined.RESULTS GCAFs displayed higher capacities to induce BGC-823 cell proliferation,migration,and invasion than GNFs(P < 0.01). Astragaloside Ⅳ treatment strongly inhibited the proliferation-,migration-and invasion-promoting capacities of GCAFs(P < 0.05 for 10 μmol/L,P < 0.01 for 20 μmol/L and 40 μmol/L). Compared with GNFs,GCAFs expressed a lower level of micro RNA-214(P < 0.01) and a higher level of micro RNA-301 a(P < 0.01). Astragaloside Ⅳ treatment significantly upregulated micro RNA-214 expression(P < 0.01) and down-regulated micro RNA-301 a expression(P < 0.01) in GCAFs. Reestablishing the micro RNA expression balance subsequently suppressed M-CSF production(P < 0.01) and secretion(P < 0.05),and elevated TIMP2 production(P < 0.01) and secretion(P < 0.05). Consequently,the ability of GCAFs to increase SOX2 and NANOG expression in BGC-823 cells was abolished by astragaloside Ⅳ.CONCLUSION Astragaloside Ⅳ can inhibit the pathological functions of GCAFs by correcting their dysregulation of micro RNA expression,and it is promisingly a potent therapeutic agent regulating tumor microenvironment.

Pharmacokinetic and pharmacodynamic analysis of ferulic acidpuerarin-astragaloside in combination with neuroprotective in cerebral ischemia/reperfusion injury in rats

Objective:To investigate the effects of the active ingredients combined therapy on inflammatory factors interleukin 1 beta(IL-1β)and neuropeptide Y(NPY)based on pharmacodynamics in rats.Methods:The animal model was built by transient middle cerebral artery occlusion(MCAO).The method for evaluating the concentrations of the FA-Pr-AI components in rat plasma was established by using HPLC and the expression levels of IL-1βand NPY were determined by ELISA.A new mathematics method of the trend of percentage rate of change(PRC)was used to assess the correlation between pharmacokinetics(PK)and pharmacodynamics(PD).Results:FA-Pr-Al in combination reduced neurological deficits,decreased infarct volume and inhibited the expression levels of IL-1βand NPY(all P<0.05)compared with the model group.FA,Pr and Al all displayed two compartment open models in rats.Clockwise hysteresis loops were obtained by time-concentration-effect curves.IL-1βand NPY level changes in the plasma followed an opposite trend to the plasma concentration tendency after C_(max)was reached.Astragaloside's PRC value was significantly higher than those of FA and puerarin between 120 to 180 min.Conclusions:The pharmacokinetics of FA-PrAl in combination were closely related its pharmacodynamics in treating ischemia/reperfusion injury,and the components of FA-Pr-Al may have a synergistic pharmacological effect.Astragaloside may play a more pronounced role in regulating IL-1βand NPY levels compared with puerarin or FA.

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

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.

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

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 has cardioprotective effect in improving cardiac function and energy metabolism through regulating lipid and glucose metabolism.The effects may be mediated by PPARγ pathway.

Content Determination of Astragaloside Ⅳ in Astragalus from Three Different Regions by HPLC

[Objectives] The content of astragaloside in Astragalus membranaceus(Fisch.) Bge.var.mongholicus(Bge.) Hisao from three different regions was determined.[Methods] Referring to the method recorded in the Chinese Pharmacopoeia(2015 edition),the content of astragaloside IV in A.membranaceus was determined by HPLC.[Results] There were great differences in the astragaloside IV content of A.membranaceus among different regions.The content of astragaloside IV in A.membranaceus cultivated in Inner Mongolia was highest(0.155%),followed by that(0.143%) in A.membranaceus cultivated in Gansu,and the content of astragaloside IV in A.membranaceus cultivated in Shanxi was lowest(0.080%).The contents of astragaloside IV in A.membranaceus from different regions were all in line with the standard(not less than 0.040%) of Chinese Pharmacopoeia(2015 edition).[Conclusions]The content of astragaloside IV in A.membranaceus cultivated in three different regions met the medicinal standards.

Molecular Mechanism of Induction on Apoptosis of Human Esophageal Cancer HCE-4 Cells by Active Components from Astragalus membranaceus

[Objectives] To investigate the pharmacologic effects of active components from A. membranaceus on human esophageal cancer HCE-4 cells and its apoptosis mechanism. [Methods] The viabilities of HCE-4 cells were measured by MTT assay. The induction of active components from A. membranaceus on apoptosis of HCE-4 cells was detected by Annexin V-FITC/PI double staining. The apoptotic-related protein expression levels were determined by Western blotting. [Results] Formononetin and astragaloside IV suppressed the proliferation of HCE-4 cells in a dose-dependent manner. The Annexin V-FITC/PI double staining results showed that formononetin and astragaloside IV could induce HCE-4 cells apoptosis in a time-dependent manner. The Western blotting results showed that formononetin and astragaloside IV could significantly down-regulate p-AKT,pro-caspase-3,and increase cle-caspase-3 protein expression in HCE-4 cells. [Conclusions]Active components from A. membranaceus such as formononetin and astragaloside IV significantly inhibited the proliferation of human esophageal cancer HCE-4 cells by inducing mitochondrial dependent apoptosis via AKT signaling pathway.

Astragaloside IV prevents high-fat diet induced obesity partially through enhancing leptin signaling transduction

Aim To investigate the effects of astragaloside IV （ASI） on high-fat diet （HFD） induced obese mice. Methods The male mice aged 6 weeks were randomly divided into three groups （u- 18/group）, namely control group, model group and ASI-treated group. Control group were fed with standard diet, whereas the other two groups were given high fat diet. ASI-treated mice were daily intraperitoneally injected with ASI （25 nag · kg^-1）. Mean- while, the other group mice were treated with saline. Body weight of mice was monitored every week and lasted for 13 weeks. Serum cholesterol and triglyceride content were measured with respective kits. Serum leptin level was deter- mined by ELISA kit. Expression of leptin receptor in hypothalamus was measured by Western blot assay. Gene ex- pression of neuropeptide Y （NPY） and agouti-related protein （AGRP） in hypothalamus was detected by qPCR assay. In addition, leptin receptor-deficient db/db mice were given intraperitoneally with ASI （25 mg ~ kg-1） or saline for 13 weeks （u- 8/group）. Results ASI blocked body weight gain, suppressed appetite, improved leptin resistance, lowered serum triacylglycerol （TG） and total cholesterol （TC） contents, reduced accumulation of fat tissues and pre- vented enlargement of adipose cells in HFD fed mice. Furthermore, ASI increased the protein expression level of lep- tin receptor in hypothalamus, and inhibited the mRNA expression levels of NPY and AGRP. However, ASI could not decrease body gain in leptin receptor - deficient db/db mice as well as the mRNA expression levels of NPY and AGRP. Conclusion The study suggested that ASI could efficiently prevent HFD-induced obesity in C57BL/6 mice,which was partially mediated through enhancing leptin signaling transduction.

Influence of astragaloside on gastric mucosa of stress ulcer rats

Objective To investigate the effect of astragaloside(AST)on the gastric mucosal injury of water immersion restraint stress ulcer rat.Methods The stress ulcer model was made by water immersion and restraint.The gastric mucosal injury index was observed.The SOD activity,the MDA contents and the gene expression of melatonin receptor 1 and 2 were detected in gastric mucosa.Results Compared with the normal group,the model group showed mucous edema,hyperemia and even ulcer damage.The injury index and the MDA content of gastric mucosa in model group were significantly increased(P<0.05),the SOD activity of gastric obviously depressed(P<0.01),and the melatonin receptor 1 and 2 mRNA expressions of damaged gastric mucosa were also lower.After administration of AST,the gastric mucosal ulcer index and MDA contents relieved obviously(P<0.01,P<0.05),the SOD activity and the expressions of melatonin receptor 1 and 2 mRNA raised up(P<0.01,P<0.05).Conclusions AST could prevent the gastric mucosal damage of rat in stress ulcer.And the mechanism of the gastric mucosal protection should be concerned with regulating the melatonin receptor and lessening the injury of oxygen free radical.

Preparation technology of Naoxueling capsules

Objective To optimize the preparation of the Naoxueling capsules.Methods To optimize the extraction of the Naoxueling prescription successively with alcohol and water,several principal ingredients in the final extracts were evaluated and data were analyzed by the orthogonal test.The capsule-molding process was investigated by measuring the angle of repose and the moisture absorption percentage.Results The optimum process of the alcohol extraction was the refluxing of herbs first in twelve-fold volume of 75% ethanol for 2 h,then in eight-fold volume of 75% ethanol for 1.5 h,and finally in seven-fold volume of 75% ethanol for 0.5 h.Meanwhile,the best water extraction was performed by boiling samples in twelve-fold amount of water for 2 h followed by another 1 h in eight-fold amount of water.To mold the capsule,the appropriate ratio of micro-powder to dextrin was 20:1.Conclusion The preparation technology of the Naoxueling capsules is reasonable and feasible,which provides evidences for the industrial manufacture.

Protective effect of astragaloside IV on cardiac hypertrophy in rats and its mechanism

Objective:To investigate the effect of astragaloside IV on cardiac hypertrophy and its regulation on autophagy.Methods:Fifty male Sprague-Dawley rats were randomly divided into sham operation group and abdominal aortic coarctation group(AAC group).There were 10 rats in sham operation group and 40 rats in the AAC group.One week after the operation,there were 32 rats in AAC group,10 rats in sham group.AAC group was randomly divided into model group,low-dose astragaloside group,high-dose astragaloside group and rapamycin group,8 rats in each group.Rapamycin group was a positive autophagy contrast agent group.They were given the corresponding solvents once a day by gavage for six weeks.At the end of study,three rats were randomly selected from each group,left ventricular mass index(LVW/BW),cardiac mass index(HW/BW)and the content of hydroxyproline were measured.HE staining,masson staining and sirius red staining were used to observe the morphological changes of myocardium.The expression of LC3II,LC3I,Beclin1,AMPK and mTOR were detected by western blot.Results:Compared with the sham operation group,AAC group showed hypertrophy,LVW/BW,HW/BW,HYP and p-mTOR/mTOR were significantly increased(P<0.05),p-AMPK/AMPK,LC3II/LC3I,Beclin1 were significantly decreased(P<0.05).Compared with the model group,the low-dose astragaloside IV group showed the hypertrophy of cardiomyocytes was relatively light,LVW/BW and HW/BW were significantly decreased(P<0.05),there was no significant difference in HYP and p-mTOR/mTOR(P>0.05),LC3II/LC3I,Beclin1 and p-AMPK/AMPK were significantly increased(P<0.05).Compared with the model group,high-dose astragaloside IV group and rapamycin group showed reduced myocardial hypertrophy,LVW/BW,HW/BW,HYP and p-mTOR/mTOR were significantly decreased(P<0.05),LC3II/LC3I,Beclin1 and p-AMPK/AMPK were significantly increased(P<0.05).Compared with the low-dose astragaloside group,the high-dose astragaloside group showed reduced myocardial hypertrophy,there were significant differences in each index(P<0.05).Compared with rapamycin group,there was no obvious difference in morphology and structure of myocardial cells,LVW/BW,HYP and p-mTOR/mTOR were decreased(P<0.05),HW/BW and p-AMPK/AMPK had no significant difference(P>0.05),LC3II/LC3I and Beclin1 were increased in high-dose astragaloside group(P<0.05).Conclusion:As IV has protective effect on cardiac hypertrophy in a dose-dependent manner and its mechanism may be related to regulate autophagy.

Determination of the Content of Astragaloside IV in Yikangshu Granules by High Performance Liquid Chromatography

[Objectives]The research aimed to establish a high performance liquid chromatography method for the content determination of astragaloside IV in Yikangshu Granules.[Methods]Kromasil 5μm C18(2),100 A,250 mm×4.6 mm was used;the mobile phase was acetonitrile-water(32∶68);flow velocity was 1.0 mL/min;the temperature of evaporator and sprayer was 80 and 30℃;the column temperature was set at 30℃,and injection volume was 20μL.[Results]Astragaloside IV showed a good linear relationship in the range of 1.01-10.14μg with the peak area,and regression equation was lgY=1.7728lgX+1.597(r=0.9999).The limit of detection for astragaloside IV was 1.96 ng,and the average recovery rate was 95.31%.[Conclusions]This method is sensitive,accurate and reproducible,with good linearity,and it is suitable for the content determination of astragaloside IV in health food Yikangshu Granules.

Astragaloside regulates the growth and invasion activity of colon cancer cells in vitro: the experimental study

Objective: To study the regulating effects of astragaloside on the growth and invasion activity of colon cancer cells in vitro. Methods: Lovo colon cancer cell lines were cultured and randomly divided into the control group treated with DMEM without drug or serum and the astragaloside groups treated with serum-free DMEM containing 10 μmol/L, 20 μmol/L and 40μmol/L astragaloside. The cell proliferation activity, the number of invasive cells as well as the expression of proliferating genes and invasion genes were measured after 24 h of treatment. Results: 24 h after treatment, the cell proliferation activity, the number of invasive cells as well as CCND1, β-catenin, ADAM17, MMP11, VEGF and ZEB2 mRNA expression of different concentrations of astragaloside groups were obviously lower than those of blank control group while WTX, PERK, ATF6, TIMP1 and E-cadherin mRNA expression were significantly higher than those of blank control group, and the higher the concentration of astragaloside, the more significant the decreasing trend of the cell proliferation activity, the number of invasive cells as well as CCND1, β-catenin, ADAM17, MMP11, VEGF and ZEB2 mRNA expression and the increasing trend of the WTX, PERK, ATF6, TIMP1 and E-cadherin mRNA expression. Conclusion: Astragaloside can significantly inhibit the growth and invasion of colon cancer cells in vitro.

Potential Role of Astragaloside IV in the Treatment of Fungal Keratitis

Fungal keratitis (FK) is a worldwide visual impairment disease. The pathogenesis of fungal keratitis involves fungi, corneal cells, inflammatory cell infiltration, collagen degradation, inflammatory cytokines and their interactions. Accumulated evidence indicated that Astragaloside IV (AS-IV) possesses a broad range of pharmacological properties, such as efficacy in anti-inflammation, alleviating fibrosis, and immunomodulatory effects. This paper summarizes new findings regarding AS-IV in immune and inflammatory diseases and analyzes the perspective application of Astragaloside IV in fungal keratitis.