Astragaloside IV (AsIV) is an active saponin extracted from Astragalus membranaceus, which has shown cardioprotective effects in a number of experimental animals. In this study we investigated the molecular mechanisms by which AsIV attenuated the myocardial ischemia reperfusion (MI/R)-induced injury in vitro and in vivo by focusing on calcium-sensing receptor (CaSR) and extracellular signal-regulated kinase 1/2 (ERK1/2). Rat neonatal cardiac myocytes were subjected to a hypoxia/reoxygenation (H/R) procedure in vitro, which significantly decreased the cell viability, increased lactate dehydrogenase (LDH) release, induced cardiomyocyte apoptosis, and increased [Ca2+]i. H/R also increased the expression of CaSR and decreased ERK1/2 phosphorylation levels in H/R-exposed myocytes. Pretreatment with AsIV (60 μmol/L) significantly improved the cell viability and decreased LDH release, attenuated myocyte apoptosis, decreased [Ca2+]i and CaSR expression, and increased the ERK1/2 phosphorylation levels. The protective effects of AsIV against H/R injury were partially inhibited by co-treatment with a CaSR agonist, gadolinium chloride (GdCl3) or with a specific ERK1/2 inhibitor U0126. For in vivo studies, a rat MI/R model was established. Pre-administration of AsIV (80 mg/kg every day, ig) significantly decreased the myocardium infarct size, creatine kinase-MB (CK-MB) production, serum cardiac troponin (cTnI) levels, and cardiomyocyte apoptosis in the rats with MI/R injury. The therapeutic effects of AsIV were associated with the downregulation of CaSR expression and upregulation of ERK1/2 phosphorylation in myocardial tissues. In summary, astragaloside IV attenuates myocardial I/R injury via inhibition of CaSR/ERK1/2 and the related apoptotic signaling pathways.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $57.00 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Heusch G. Cardioprotection: chances and challenges of its translation to the clinic. Lancet. 2013;381:166–75.
Levitsky S. Protecting the myocardial cell during coronary revascularization. The William W. L. Glenn Lecture. Circulation. 2006;114:I339–43.
Yellon DM, Hausenloy DJ. Myocardial reperfusion injury. N Engl J Med. 2007;357:1121–35.
Zhou X, Sun X, Gong X, Yang Y, Chen C, Shan G, et al. Astragaloside IV from Astragalus membranaceus ameliorates renal interstitial fibrosis by inhibiting inflammation via TLR4/NF-small ka, CyrillicB in vivo and in vitro. Int Immunopharmacol. 2017;42:18–24.
Guo H, Cao A, Chu S, Wang Y, Zang Y, Mao X, et al. Astragaloside IV attenuates podocyte apoptosis mediated by endoplasmic reticulum stress through upregulating sarco/endoplasmic reticulum Ca2+-ATPase 2 expression in diabetic nephropathy. Front Pharmacol. 2016;7:500.
Jiang XG, Sun K, Liu YY, Yan L, Wang MX, Fan JY, et al. Astragaloside IV ameliorates 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis implicating regulation of energy metabolism. Sci Rep. 2017;7:41832.
Xu C, Tang F, Lu M, Yang J, Han R, Mei M, et al. Pretreatment with astragaloside IV protects human umbilical vein endothelial cells from hydrogen peroxide induced oxidative stress and cell dysfunction via inhibiting eNOS uncoupling and NADPH oxidase - ROS - NF-kappaB pathway. Can J Physiol Pharmacol. 2016;94:1–9.
Zhao M, Zhao J, He G, Sun X, Huang X, Hao L. Effects of astragaloside IV on action potentials and ionic currents in guinea-pig ventricular myocytes. Biol Pharm Bull. 2013;36:515–21.
Lu M, Wang H, Wang J, Zhang J, Yang J, Liang L, et al. Astragaloside IV protects against cardiac hypertrophy via inhibiting the Ca2+/CaN signaling pathway. Planta Med. 2014;80:63–9.
Zhang S, Tang F, Yang Y, Lu M, Luan A, Zhang J, et al. Astragaloside IV protects against isoproterenol-induced cardiac hypertrophy by regulating NF-kappaB/PGC-1alpha signaling mediated energy biosynthesis. PLoS One. 2015;10:e0118759.
Zhang WD, Chen H, Zhang C, Liu RH, Li HL, Chen HZ. Astragaloside IV from Astragalus membranaceus shows cardioprotection during myocardial ischemia in vivo and in vitro. Planta Med. 2006;72:4–8.
Tu L, Pan CS, Wei XH, Yan L, Liu YY, Fan JY, et al. Astragaloside IV protects heart from ischemia and reperfusion injury via energy regulation mechanisms. Microcirculation. 2013;20:736–47.
Xu XL, Chen XJ, Ji H, Li P, Bian YY, Yang D, et al. Astragaloside IV improved intracellular calcium handling in hypoxia-reoxygenated cardiomyocytes via the sarcoplasmic reticulum Ca-ATPase. Pharmacology. 2008;81:325–32.
Tfelt-Hansen J, Hansen JL, Smajilovic S, Terwilliger EF, Haunso S, Sheikh SP. Calcium receptor is functionally expressed in rat neonatal ventricular cardiomyocytes. Am J Physiol Heart Circ Physiol. 2006;290:H1165–71.
Sun YH, Liu MN, Li H, Shi S, Zhao YJ, Wang R, et al. Calcium-sensing receptor induces rat neonatal ventricular cardiomyocyte apoptosis. Biochem Biophys Res Commun. 2006;350:942–8.
Li HX, Kong FJ, Bai SZ, He W, Xing WJ, Xi YH, et al. Involvement of calcium-sensing receptor in oxLDL-induced MMP-2 production in vascular smooth muscle cells via PI3K/Akt pathway. Mol Cell Biochem. 2012;362:115–22.
Jiang CM, Han LP, Li HZ, Qu YB, Zhang ZR, Wang R, et al. Calcium-sensing receptors induce apoptosis in cultured neonatal rat ventricular cardiomyocytes during simulated ischemia/reperfusion. Cell Biol Int. 2008;32:792–800.
Lu F, Tian Z, Zhang W, Zhao Y, Bai S, Ren H, et al. Calcium-sensing receptors induce apoptosis in rat cardiomyocytes via the endo(sarco)plasmic reticulum pathway during hypoxia/reoxygenation. Basic Clin Pharmacol Toxicol. 2010;106:396–405.
Zheng H, Liu J, Liu C, Lu F, Zhao Y, Jin Z, et al. Calcium-sensing receptor activating phosphorylation of PKCdelta translocation on mitochondria to induce cardiomyocyte apoptosis during ischemia/reperfusion. Mol Cell Biochem. 2011;358:335–43.
Kabir ME, Singh H, Lu R, Olde B, Leeb-Lundberg LM, Bopassa JC. G protein-coupled estrogen receptor 1 mediates acute estrogen-induced cardioprotection via MEK/ERK/GSK-3beta pathway after ischemia/reperfusion. PLoS One. 2015;10:e0135988.
Filippone SM, Samidurai A, Roh SK, Cain CK, He J, Salloum FN, et al. Reperfusion therapy with rapamycin attenuates myocardial infarction through activation of AKT and ERK. Oxid Med Cell Longev. 2017;2017:4619720.
Wu G, Wang H, Yang J, Yang Y, Liu C, Jing L, et al. Kappa-Opioid receptor stimulation inhibits augmentation of Ca2+ transient and hypertrophy induced by isoprenaline in neonatal rat ventricular myocytes—role of CaMKIIdelta(B). Eur J Pharmacol. 2008;595:52–7.
Yin Y, Guan Y, Duan J, Wei G, Zhu Y, Quan W, et al. Cardioprotective effect of Danshensu against myocardial ischemia/reperfusion injury and inhibits apoptosis of H9c2 cardiomyocytes via Akt and ERK1/2 phosphorylation. Eur J Pharmacol. 2013;699:219–26.
Guo J, Li HZ, Zhang WH, Wang LC, Wang LN, Zhang L, et al. Increased expression of calcium-sensing receptors induced by ox-LDL amplifies apoptosis of cardiomyocytes during simulated ischaemia-reperfusion. Clin Exp Pharmacol Physiol. 2010;37:e128–35.
Greenberg HZE, Jahan KS, Shi J, Vanessa Ho WS, Albert AP. The calcilytics Calhex-231 and NPS 2143 and the calcimimetic Calindol reduce vascular reactivity via inhibition of voltage-gated Ca2+ channels. Eur J Pharmacol. 2016;791:659–68.
Zhang Y, Chen G, Zhong S, Zheng F, Gao F, Chen Y, et al. N-n-butyl haloperidol iodide ameliorates cardiomyocytes hypoxia/reoxygenation injury by extracellular calcium-dependent and -independent mechanisms. Oxid Med Cell Longev. 2013;2013:912310.
Hadi NR, Al-Amran F, Yousif M, Zamil ST. Antiapoptotic effect of simvastatin ameliorates myocardial ischemia/reperfusion injury. ISRN Pharmacol. 2013;2013:815094.
Lu M, Tang F, Zhang J, Luan A, Mei M, Xu C, et al. Astragaloside IV attenuates injury caused by myocardial ischemia/reperfusion in rats via regulation of toll-like receptor 4/nuclear factor-kappaB signaling pathway. Phytother Res. 2015;29:599–606.
Si J, Wang N, Wang H, Xie J, Yang J, Yi H, et al. HIF-1alpha signaling activation by post-ischemia treatment with astragaloside IV attenuates myocardial ischemia-reperfusion injury. PLoS One. 2014;9:e107832.
Han XH, Liu P, Zhang YY, Zhang N, Chen FR, Cai JF. Astragaloside IV regulates expression of ATP-sensitive potassium channel subunits after ischemia-reperfusion in rat ventricular cardiomyocytes. J Tradit Chin Med. 2011;31:321–6.
Li ZP, Cao Q. Effects of astragaloside IV on myocardial calcium transport and cardiac function in ischemic rats. Acta Pharmacol Sin. 2002;23:898–904.
Fliss H, Gattinger D. Apoptosis in ischemic and reperfused rat myocardium. Circ Res. 1996;79:949–56.
Hu W, Zhang P, Gu J, Yu Q, Zhang D. NEDD4-1 protects against ischaemia/reperfusion-induced cardiomyocyte apoptosis via the PI3K/Akt pathway. Apoptosis. 2017;22:437–48.
Zhang SW, Liu Y, Wang F, Qiang J, Liu P, Zhang J, et al. Ilexsaponin A attenuates ischemia-reperfusion-induced myocardial injury through anti-apoptotic pathway. PLoS One. 2017;12:e0170984.
Brown EM, Gamba G, Riccardi D, Lombardi M, Butters R, Kifor O, et al. Cloning and characterization of an extracellular Ca2+-sensing receptor from bovine parathyroid. Nature. 1993;366:575–80.
Wang R, Xu C, Zhao W, Zhang J, Cao K, Yang B, et al. Calcium and polyamine regulated calcium-sensing receptors in cardiac tissues. Eur J Biochem. 2003;270:2680–8.
Guo J, Li HZ, Wang LC, Zhang WH, Li GW, Xing WJ, et al. Increased expression of calcium-sensing receptors in atherosclerosis confers hypersensitivity to acute myocardial infarction in rats. Mol Cell Biochem. 2012;366:345–54.
Lu FH, Fu SB, Leng X, Zhang X, Dong S, Zhao YJ, et al. Role of the calcium-sensing receptor in cardiomyocyte apoptosis via the sarcoplasmic reticulum and mitochondrial death pathway in cardiac hypertrophy and heart failure. Cell Physiol Biochem. 2013;31:728–43.
Sun J, Murphy E. Calcium-sensing receptor: a sensor and mediator of ischemic preconditioning in the heart. Am J Physiol Heart Circ Physiol. 2010;299:H1309–17.
Irving EA, Bamford M. Role of mitogen- and stress-activated kinases in ischemic injury. J Cereb Blood Flow Metab. 2002;22:631–47.
Strohm C, Barancik T, Bruhl ML, Kilian SA, Schaper W. Inhibition of the ER-kinase cascade by PD98059 and UO126 counteracts ischemic preconditioning in pig myocardium. J Cardiovasc Pharmacol. 2000;36:218–29.
Wang P, Wang L, Wang S, Li S, Li Y, Zhang L. Effects of calcium-sensing receptors on apoptosis in rat hippocampus during hypoxia/reoxygenation through the ERK1/2 pathway. Int J Clin Exp Pathol. 2015;8:10808–15.
Ye J, Ai W, Zhang F, Zhu X, Shu G, Wang L, et al. Enhanced proliferation of porcine bone marrow mesenchymal stem cells induced by extracellular calcium is associated with the activation of the calcium-sensing receptor and ERK signaling pathway. Stem Cells Int. 2016;2016:6570671.
This work was supported by the National Natural Science Foundation of China (No 81703739), the Natural Science Foundation of Liaoning Province (No 201602284), and the Innovative Training Program for College Students of Liaoning Province (No 201610160000040).
B.Y. and M.-i.L. designed the research. B.Y., X.-w.H. and M.-i.L. performed the research. B.Y. analyzed the data. M.-i.L. wrote the paper.
The authors declare no competing interests.
About this article
Astragaloside IV Protects Ethanol-Induced Gastric Mucosal Injury by Preventing Mitochondrial Oxidative Stress and the Activation of Mitochondrial Pathway Apoptosis in Rats
Frontiers in Pharmacology (2019)
Creative surgery and oncology (2019)
Frontiers in Pharmacology (2019)
Protection of the myocardium against ischemia/reperfusion injury by punicalagin through an SIRT1-NRF-2-HO-1-dependent mechanism
Chemico-Biological Interactions (2019)
Chemical diversity and biological activities of the saponins isolated from Astragalus genus: focus on Astragaloside IV
Phytochemistry Reviews (2019)