Anti-inflammatory and DNA Repair Effects of Astragaloside IV on PC12 Cells Damaged by Lipopolysaccharide

Objective This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide(LPS)and to examine the effects of astragaloside IV on key targets using high-throughput sequence technology and bioinformatics analyses.Methods PC12 cells in the logarithmic growth phase were treated with LPS at final concentrations of 0.25,0.5,0.75,1,and 1.25 mg/mL for 24 h.Cell morphology was evaluated,and cell survival rates were calculated.A neurocyte inflammatory model was established with LPS treatment,which reached a 50%cell survival rate.PC12 cells were treated with 0.01,0.1,1,10,or 100µmol/L astragaloside IV for 24 h.The concentration of astragaloside IV that did not affect the cell survival rate was selected as the treatment group for subsequent experiments.NOS activity was detected by colorimetry;the expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS and COX-2 mRNA and protein were detected by RT-qPCR and Western blotting.The differentially expressed genes(DEGs)between the groups were screened using a second-generation sequence(fold change>2,P<0.05)with the following KEGG enrichment analysis,RT-qPCR and Western blotting were used to detect the mRNA and protein expression of DEGs related to the IL-17 pathway in different groups of PC12 cells.Results The viability of PC12 cells was not altered by treatment with 0.01,0.1,or 1µmol/L astragaloside IV for 24 h(P>0.05).However,after treatment with 0.5,0.75,1,or 1.25 mg/mL LPS for 24 h,the viability steadily decreased(P<0.01).The mRNA and protein expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS,and COX-2 were significantly increased after PC12 cells were treated with 1 mg/mL LPS for 24 h(P<0.01);however,these changes were reversed when PC12 cells were pretreated with 0.01,0.1,or 1µmol/L astragaloside IV in PC12 cells and then treated with 1 mg/mL LPS for 24 h(P<0.05).Second-generation sequencing revealed that 1026 genes were upregulated,while 1287 genes were downregulated.The DEGs were associated with autophagy,TNF-α,interleukin-17,MAPK,P53,Toll-like receptor,and NOD-like receptor signaling pathways.Furthermore,PC12 cells treated with a 1 mg/mL LPS for 24 h exhibited increased mRNA and protein expression of CCL2,CCL11,CCL7,MMP3,and MMP10,which are associated with the IL-17 pathway.RT-qPCR and Western blotting analyses confirmed that the DEGs listed above corresponded to the sequence assay results.Conclusion LPS can damage PC12 cells and cause inflammatory reactions in nerve cells and DNA damage.astragaloside IV plays an anti-inflammatory and DNA damage protective role and inhibits the IL-17 signaling pathway to exert a neuroprotective effect in vitro.

Astragaloside Ⅳ Protects Against Aβ1-42-induced Oxidative Stress, Neuroinflammation and Cognitive Impairment in Rats

Objective To investigate the neuroprotective action of astragaloside Ⅳ(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-Ⅳ groups. 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 8 th 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 softwaresystem. The vision and swim speeds of rats were also determined to exclude the effect of these factors on the parameters of learning and memory. After behavioral tests, the rats were sacrificed immediately by decapitation. Hippocampus were collected. The enzyme activities of superoxide dismutase(SOD), glutathione peroxidase(GSH-px) and catalase(CAT) in the hippocampus obtained from different-treated rat brain were measured by following the manufacturer’s instructions. The levels of interleukin-1 beta(IL-1β) and tumor necrosis factor-alpha(TNF-α) in tissue lysates were assayed with ELISA.Results The water maze test results indicated that chronic treatments with AS-Ⅳ effectively protected the rats from Aβ1-42-induced spatial learning and memory impairment. Furthermore, the activities of SOD, GSH-px and CAT decreased by Aβ1-42 were also restored by AS-Ⅳ treatment in the hippocampus of rats. In addition, AS-Ⅳ significantly decreased the levels of IL-1β and TNF-α in the hippocampus of Aβ1-42-induced amnesia’s rats. Conclusion Our findings suggest that AS-Ⅳ might be a useful chemical in improving the spatial memory and relieving the oxidative stress and neuroinflammation in Alzheimer patients.

Determination of Astragaloside IV in Yupingfeng Oral Solution by HPLC-ELSD Method

[Objective] This study aimed to establish a method for determining the content of Astragaloside IV in Yupingfeng oral solution.[Method] The HPLC-ELSD method was adopted.The chromatographic column was Venusil MP（4.6 mm × 150 mm,5 μm）.The mobile phase was acetonitrile-water（35∶65）.The ELSD evaporator tube temperature was 65 ℃.N2 was used as the carrier gas（pressure,30 psi）.[Result] When the content of Astragaloside IV ranged from 0.5 to 5.0 μg,the Astragaloside IV content showed a good linear relationship with peak area（r=0.999,n=6）.The average recovery was 96.36%,and the RSD was 2.46%.[Conclusion] This method is accurate and reliable,and can be applied in the quality control of Yupingfeng oral solution.

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.

The mechanism of astragaloside Ⅳ promoting sciatic nerve regeneration

3-O-beta-D-xylopyranosyl-6-O-beta-D-glucopyranosyl-cycloastragenol （astragaloside IV）, the main active component of the traditional Chinese medicine astragalus membranaceus, has been shown to be neuroprotective. This study investigated whether astragaloside IV could promote the repair of injured sciatic nerve. Denervated sciatic nerve of mice was subjected to anastomosis. The mice were intraperitoneally injected with 10, 5, 2.5 mg/kg astragaloside IV per day for 8 consecutive days Western blot assay and real-time PCR results demonstrated that growth-associated protein-43 ex- pression was upregulated in mouse spinal cord segments L4-6 after intervention with 10, 5, 2.5 mg/kg astragaloside IV per day in a dose-dependent manner. Luxol fast blue staining and elec- trophysiological detection suggested that astragaloside IV elevated the number and diameter of myelinated nerve fibers, and simultaneously increased motor nerve conduction velocity and action potential amplitude in the sciatic nerve of mice. These results indicated that astragaloside IV con- tributed to sciatic nerve regeneration and functional recovery in mice. The mechanism underlying this effect may be associated with the upregulation of growth-associated protein-43 expression.

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.

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.

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.

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.

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.

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.

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.

Angiogenesis function of astragaloside Ⅳ in rats with myocardial infarction via PKD1-HDAC5-VEGF pathway

OBJECTⅣE This study aimed to investigate the role and mechanism of Astragaloside Ⅳ(AS-Ⅳ) in rats with myocardial infarction.METHODS The myocardial infarction model was established by ligation of the left anterior descending artery.The rats were randomly divided into sham,DMSO,model group,AS-Ⅳ and CID755673 groups.The rats were sacrificed 4 weeks later,and segmental heart samples were used for hematoxylin and eosin staining and masson staining.The expression of PKD1,HDAC5 and VEGF were analyzed using immunohistochemistry,reverse transcription poly.merase chain reaction and western blot.RESULTS Compared with the sham operation and DMSO groups,morphology of myocardium in model group was disordered,accompanied with necrotic myocar.dial cells and obvious collagen tissues.After treatment with AS-Ⅳ,the morphology of myocardium was obviously improved,and the number of new blood vessels increased significantly.However,after treatment with CID755673,the myocardial tissue of rats became disordered again,the necrotic cells increased,and some vessels closed.The expression levels of PKD1,HDAC5 and VEGF mRNA and protein in myocardial tissue of model group were significantly lower than the other four groups(P<0.05),whereas these levels in the AS-Ⅳ group were significantly higher than those in the other four groups(P<0.01).Additionally,the CID755673 group had significantly higher levels of PKD1,HDAC5 and VEGF mRNA and protein than the sham group,DMSO group and model group(P<0.05).CONCLUSION AS-Ⅳ may partly promote the angiogenesis of myocardial tissue in rats with myocardial infarction via the PKD1-HDAC5-VEGF pathway.

An updated role of astragaloside Ⅳ in heart failure

Heart failure has become a global public health problem that seriously threatens human health.Due to "multi-target" effect,traditional Chinese medicine has unique advantages in the treat.ment of heart failure.Astragaloside Ⅳ is one of the main active ingredients of astragalus membrana.ceus.It has the functions of protecting the heart and neovascularization,anti-inflammation,anti-oxida.tion,regulating energy metabolism,neuroprotection and anti-cancer effects.This article reviews the recent progresses of astragaloside Ⅳ in the treatment of heart failure.Data show that astragaloside Ⅳ can inhibit heart fibrosis,attenuate excessively activation of renin-angiotensin system,increase energy metabolism,promote positive inotropic action,inhibit myocardial cell apoptosis,etc,which are all involved in the protective role of astragalosideⅣ in rodents or cellular models of heart failure.Astragalo.side Ⅳ may be a potential active ingredient in traditional Chinese medicine for the treatment of heart failure.

Study on mechanism of astragaloside Ⅳ in treatment of pulmonary fibrosis with epithelial-mesenchymal transition

OBJECTIVE Currently discussing the clinical treatment of pulmonary fibrosis commonly used drugs astragalus main active ingredient astragalosideⅣ(ASTⅣ) in vitro after transforming growth factor-β1 induced lung adenocarcinoma A549 epithelial cells after epithelial cell interstitial EpithelialMesenchymal Transition(EMT).METHODS The effect of astragalosideIV on the proliferation of A549 cells was detected by MTT assay for the first time.No significant effect of astragaloside on the prolifera.tion of A549 cells was found in the range of 1.25-20 μmol/L.A549 cells in vitro were divided into 5 groups:normal group,control group,low,medium and high experimental groups,which were treated for 72 hours,and the morphological changes of cells in each group were observed by light microscope.Real-time quantitative PCR(qPCR) and Western blotting were performed.Detection of gene and protein expression levels.RESULTS The results of real-time fluorescence quantitative PCR showed that the quantitative analysis of high-dose astragalosideⅣ in the experimental group was lower than that of the control group in the α-SMA analysis,and the difference was statistically significant(P<0.05).The dose of Astragaloside Ⅳ in the experimental group was higher than that of the control group in the E-Cad analysis,and the difference was statistically significant(P<0.05).Western Blot results showed that the expression of α-SMA antibody in the high-dose and low-dose experimental group was lower than that in the control group,the difference was statistically significant(P<0.05).The high-dose experimental group had a significantly higher expression of E-Cad antibody than the control group,the difference was statistically significant(P<0.01).CONCLUSION This study uses A549 epithelial cells as a model,A549 cells were modeled and confirmed that Astragaloside can effectively inhibit TGF-β1-induced epithelialmesenchymal transition(EMT) and provide a new basis for the treatment of pulmonary fibrosis.