Isorhynchophylline (IRN) is one of the major tetracyclic oxindole alkaloids found in Uncaria rhynchophylla. Studies have found that IRN has diverse biological activities including antioxidant, anti-apoptosis, and neuroprotection. However, little is known about the effect of IRN on the development of cardiac hypertrophy. In this study, we investigated the change of the cell surface area and nascent protein synthesis of cultured H9c2 cardiomyocytes on exposure to phenylephrine (PE) plus IRN, and thus confirmed that IRN ameliorated cardiomyocyte hypertrophy induced by PE in vitro. Meanwhile, it turns out that IRN is also effective in neonatal rat ventricular myocytes (NRVMs) stimulated with angiotensin II (AngII). We also showed that IRN prevented cardiac dysfunction in mice with pressure overload due to transverse aortic constriction (TAC) and attenuated cardiac hypertrophy and fibrosis. IRN treatment improved the cardiac function assessed by echocardiographic parameters fractional shortening (FS) as well as suppressed the cardiac hypertrophy phenotypes, such as the increasing of ventricular mass/body weight and myocyte cross-sectional area. RT-PCR analysis showed that IRN treatment also alleviated the expression of fetal genes of ANP, BNP, Myh7, and the correlated fibrosis genes including TGF-β1, collagen I, collagen III, and CTGF in vivo. Meanwhile, IRN had anti-oxidative effects on cardiac remodeling with suppressed 4-HNE and MDA. Western blot analysis showed that the Nrf2 nuclear translocation and MAPK pathway were involved in the potential mechanisms of IRN on cardiac hypertrophy inhibition. The results of our study provide further evidence that IRN is a promising drug for the treatment of cardiac hypertrophy.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Barry SP, Davidson SM, Townsend PA (2008) Molecular regulation of cardiac hypertrophy. Int J Biochem Cell Biol 40:2023–2039
Bernardo BC, Weeks KL, Pretorius L, McMullen JR (2010) Molecular distinction between physiological and pathological cardiac hypertrophy: experimental findings and therapeutic strategies. Pharmacol Ther 128:191–227
Chen QM, Maltagliati AJ (2018) Nrf2 at the heart of oxidative stress and cardiac protection. Physiol Genomics 50:77–97
Cui S, Cui Y, Li Y, Zhang Y, Wang H, Qin W, Chen X, Ding S, Wu D, Guo H (2018) Inhibition of cardiac hypertrophy by aromadendrin through down-regulating NFAT and MAPKs pathways. Biochem Biophys Res Commun 506:805–811
Dong B, Xue R, Sun Y, Dong Y, Liu C (2017) Sestrin 2 attenuates neonatal rat cardiomyocyte hypertrophy induced by phenylephrine via inhibiting ERK1/2. Mol Cell Biochem 433:113–123
Fan C, Li Y, Yang H, Cui Y, Wang H, Zhou H, Zhang J, du B, Zhai Q, Wu D, Chen X, Guo H (2019) Tamarixetin protects against cardiac hypertrophy via inhibiting NFAT and AKT pathway. J Mol Histol 50:343–354
Gan R, Dong G, Yu J, Wang X, Fu S, Yang S (2011) Protective effects of isorhynchophylline on cardiac arrhythmias in rats and Guinea pigs. Planta Med 77:1477–1481
Guo H, Zhang X, Cui Y, Deng W, Xu D, Han H, Wang H, Chen Y, Li Y, Wu D (2014) Isorhynchophylline protects against pulmonary arterial hypertension and suppresses PASMCs proliferation. Biochem Biophys Res Commun 450:729–734
Guo H, Zhang X, Cui Y, Zhou H, Xu D, Shan T, Zhang F, Guo Y, Chen Y, Wu D (2015) Taxifolin protects against cardiac hypertrophy and fibrosis during biomechanical stress of pressure overload. Toxicol Appl Pharmacol 287:168–177
Guo W, Zhu H, Wang Z, Chen JA, Wu J, Zhu Y, Gu X (2017) Novel rhynchophylline analogues as microvascular relaxation agents for the treatment of microvascular dysfunction caused by diabetes. Eur J Med Chem 139:657–664
Howden R (2013) Nrf2 and cardiovascular defense. Oxidative Med Cell Longev 2013:104308
Kawakami Z, Kanno H, Ikarashi Y, Kase Y (2011) Yokukansan, a kampo medicine, protects against glutamate cytotoxicity due to oxidative stress in PC12 cells. J Ethnopharmacol 134:74–81
Kehat I, Molkentin JD (2010) Molecular pathways underlying cardiac remodeling during pathophysiological stimulation. Circulation 122:2727–2735
Lee H, Baek SH, Lee JH, Kim C, Ko JH, Lee SG, Chinnathambi A, Alharbi SA, Yang WM, Um JY, Sethi G, Ahn KS (2017) Isorhynchophylline, a potent plant alkaloid, induces apoptotic and anti-metastatic effects in human hepatocellular carcinoma cells through the modulation of diverse cell signaling cascades. Int J Mol Sci 18(5)
Li J, Zhang C, Xing Y, Janicki JS, Yamamoto M, Wang XL, Tang DQ, Cui T (2011) Up-regulation of p27(kip1) contributes to Nrf2-mediated protection against angiotensin II-induced cardiac hypertrophy. Cardiovasc Res 90:315–324
Li HQ, Ip SP, Zheng GQ, Xian YF, Lin ZX (2018a) Isorhynchophylline alleviates learning and memory impairments induced by aluminum chloride in mice. Chin Med 13:29
Li Q, Niu C, Zhang X, Dong M (2018b) Gastrodin and Isorhynchophylline synergistically inhibit MPP(+)-induced oxidative stress in SH-SY5Y cells by targeting ERK1/2 and GSK-3beta pathways: involvement of Nrf2 nuclear translocation. ACS Chem Neurosci 9:482–493
Liu R, Molkentin JD (2016) Regulation of cardiac hypertrophy and remodeling through the dual-specificity MAPK phosphatases (DUSPs). J Mol Cell Cardiol 101:44–49
Ma Q, Liu Y, Chen L (2018) JIP3 deficiency attenuates cardiac hypertrophy by suppression of JNK pathway. Biochem Biophys Res Commun 503:1–7
Martin TP, Robinson E, Harvey AP, MacDonald M, Grieve DJ, Paul A, Currie S (2012) Surgical optimization and characterization of a minimally invasive aortic banding procedure to induce cardiac hypertrophy in mice. Exp Physiol 97:822–832
Maulik SK, Kumar S (2012) Oxidative stress and cardiac hypertrophy: a review. Toxicol Mech Methods 22:359–366
Moris D, Spartalis M, Tzatzaki E et al (2017) The role of reactive oxygen species in myocardial redox signaling and regulation. Ann Transl Med 5:324
Mutlak M, Schlesinger-Laufer M, Haas T, Shofti R, Ballan N, Lewis YE, Zuler M, Zohar Y, Caspi LH, Kehat I (2018) Extracellular signal-regulated kinase (ERK) activation preserves cardiac function in pressure overload induced hypertrophy. Int J Cardiol 270:204–213
Narasimhan M, Rajasekaran NS (2016) Exercise, Nrf2 and antioxidant signaling in cardiac aging. Front Physiol 7:241
Petrich BG, Eloff BC, Lerner DL, Kovacs A, Saffitz JE, Rosenbaum DS, Wang Y (2004) Targeted activation of c-Jun N-terminal kinase in vivo induces restrictive cardiomyopathy and conduction defects. J Biol Chem 279:15330–15338
Wu JB, Zhou Y, Liang CL, Zhang XJ, Lai JM, Ye SF, Ouyang H, Lin J, Zhou JY (2017) Cyclovirobuxinum D alleviates cardiac hypertrophy in hyperthyroid rats by preventing apoptosis of cardiac cells and inhibiting the p38 mitogen-activated protein kinase signaling pathway. Chin J Integr Med 23:770–778
Yuan D, Ma B, Yang JY, Xie YY, Wang L, Zhang LJ, Kano Y, Wu CF (2009) Anti-inflammatory effects of rhynchophylline and isorhynchophylline in mouse N9 microglial cells and the molecular mechanism. Int Immunopharmacol 9:1549–1554
Zeisberg EM, Tarnavski O, Zeisberg M, Dorfman AL, McMullen JR, Gustafsson E, Chandraker A, Yuan X, Pu WT, Roberts AB, Neilson EG, Sayegh MH, Izumo S, Kalluri R (2007) Endothelial-to-mesenchymal transition contributes to cardiac fibrosis. Nat Med 13:952–961
Zhang F, Sun AS, Yu LM, Wu Q, Gong QH (2008) Effects of isorhynchophylline on angiotensin II-induced proliferation in rat vascular smooth muscle cells. J Pharm Pharmacol 60:1673–1678
Zhang Y, Fang Liu F, Bi X, Wang S, Wu X, Jiang F (2015) The antioxidant compound tert-butylhydroquinone activates Akt in myocardium, suppresses apoptosis and ameliorates pressure overload-induced cardiac dysfunction. Sci Rep 5:13005
Zhou JY, Zhou SW (2012) Isorhynchophylline: a plant alkaloid with therapeutic potential for cardiovascular and central nervous system diseases. Fitoterapia 83:617–626
This work was supported by grants from the National Natural Science Foundation of China (No. 81300459; No. 81300162), Natural Science Foundation of Shandong Province (No. ZR2016HM48). Project funded by China Postdoctoral Science Foundation (No. 2018T110694; No. 2017M610429; No. 2014M551921) and the International Postdoctoral Exchange Fellowship Program.
Conflict of interest
The authors declare that they have no conflicts of interest.
All animal experiments were performed in accordance with the guidelines for the care and use of laboratory animals, published by the Ministry of the People’s Republic of China, and were approved by the Institutional Animal Care and Use Committee of Shandong University.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
IRN attenuated protein synthesis and hypertrophy of NRVMs induced by AngII. a IRN alleviated the increased protein synthesis after treated by AngII. Cells were treated with or without IRN (25 μM) for 48 h. *P < 0.05 vs. corresponding control; #P < 0.05 vs. 1 μM AngII. b Real-time PCR results showing the mRNA levels of BNP and ANP genes after AngII treatment. *P < 0.05 vs. corresponding control; #P < 0.05 vs. 1 μM AngII. c The representative pictures of α-Actinin immunofluorescence staining. d The quantification of cardiomyocyte sizes after AngII and IRN treatment. All experiments were repeated 3 times. *P < 0.05 vs. corresponding control; #P < 0.05 vs. 1 μM AngII. (PDF 104 kb)
Effect of IRN on Nrf2 and MAPKs signaling pathways in NRVMs stimulated with AngII. a, b Western blotting results of nuclear Nrf2 change in NRVMs. c–f Western blotting results of the total and phosphorylated protein levels of ERK1/2, P38 MAPK, and JNK. All experiments were repeated three times. *P < 0.05 vs. corresponding control; #P < 0.05 vs. AngII group. (PDF 163 kb)
Nrf2 inhibition abolishes protective effect of IRN on cardiac hypertrophy in NRVMs. NRVMs transfected with (50 nM) Nrf2-siRNA and control-siRNA were treated or not with AngII 1 μM and IRN 25 μM for 48 h. a–c Effects of IRN, Nrf2-siRNA, and control-siRNA on the activation of Nrf2 and production of ROS in AngII-treated NRVMs. d Effects of IRN 25 μM, Nrf2-siRNA, and control-siRNA on the cardiomyocyte area in AngII-treated NRVMs. All experiments were repeated three times. *P < 0.05 vs. control group; #P < 0.05 vs. AngII + IRN group. (PDF 90 kb)
Isorhynchophylline enhances Nrf2 and inhibits MAPK pathway in cardiac hypertrophy. A potential working model of IRN regulating oxidative stress, Nrf2 nuclear translocation and activation of MAPKs in cardiac hypertrophy. (PDF 63 kb)
(DOC 100 kb)
About this article
Cite this article
Zhang, Y., Cui, Y., Dai, S. et al. Isorhynchophylline enhances Nrf2 and inhibits MAPK pathway in cardiac hypertrophy. Naunyn-Schmiedeberg's Arch Pharmacol 393, 203–212 (2020). https://doi.org/10.1007/s00210-019-01716-0
- Cardiac hypertrophy
- Oxidative stress