Role of MAPK/NF-κB pathway in cardioprotective effect of Morin in isoproterenol induced myocardial injury in rats

  • Vipin Kumar Verma
  • Salma Malik
  • Susrutha P. Narayanan
  • Ekta Mutneja
  • Anil Kumar Sahu
  • Jagriti Bhatia
  • Dharamvir Singh AryaEmail author
Original Article


Oxidative stress plays a major role in myocardial injury. Morin, a bioflavonoid has known to possess various biological activities in previous studies. Hence, this study evaluated the cardioprotective mechanism(s) of Morin against isoproterenol induced myocardial necrosis in rats. Male albino Wistar rats were divided into five groups (n = 8) i.e., I (normal), II (ISO-control), III, IV and V (morin 20, 40 and 80 mg/kg respectively). Groups III, IV and V were treated orally with daily doses of Morin accordingly for 28 days. On 26th and 27th day, a single injection of isoproterenol was injected (85 mg/kg s.c.) at 24 h interval to induce myocardial necrosis in group II, III, IV and V. On 28th day, hemodynamic parameters were evaluated, animals were euthanised and heart was excised for measurement of various parameters. In ISO-control rats, there was deterioration of hemodynamic parameters, decreased anti-oxidants levels, increased cardiac injury markers and pro-inflammatory cytokines (TNF-α and IL-6). Also, there was increased level of Bax, Caspase-3, p-JNK, p-38 and NF-κB and decreased expression of Bcl-2 and p-ERK1/2 in ISO-C group. Morin dose-dependently improved hemodynamic profile, increased anti-oxidant levels, normalized myocardial architecture and reduced inflammatory markers and apoptosis. Furthermore, immunoblot analysis of MAPK pathway proteins demonstrated the mechanism responsible for anti-apoptotic and anti-inflammatory potential of morin. Thus, this study substantiated the beneficial effect of Morin by virtue of its modulation of MAPK pathway in myocardial injury.


Morin Myocardial infarction Oxidative stress Apoptosis MAPK/ERK pathway 



All authors are thankful to the technical staff in cardiovascular pharmacology lab. Vipin Kumar Verma is also thankful to DST-SERB to provide fellowship and financial assistance to conduct experiments.


The funding was provided by Department of Science and Technology, Government of India (Grant Number PDF/2016/003885).

Compliance with ethical standards

Conflict of interest

Authors have no conflict of interest to declare.

Research involving in human and animal rights

All experimental procedures were performed in accordance with the protocol approved by Institutional Animal Ethics Committee, AIIMS for the file number 03/IAEC-1/2017 (Registration No. 10/GO/ReBi/S/99/CPCSEA).


  1. 1.
    Lozano R, Naghavi M, Foreman K et al (2012) Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380:2095–2128CrossRefGoogle Scholar
  2. 2.
    Murray CJL, Vos T, Lozano R et al (2012) Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990–2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380:2197–2223CrossRefGoogle Scholar
  3. 3.
    Tanwar V, Sachdeva J, Kishore K et al (2010) Dose-dependent actions of curcumin in experimentally induced myocardial necrosis: a biochemical, histopathological, and electron microscopic evidence. Cell Biochem Funct 28:74–82CrossRefGoogle Scholar
  4. 4.
    Vishwakarma A, Singh TU, Rungsung S et al (2018) Effect of Kaempferol pretreatment on myocardial injury in rats. Cardiovasc Toxicol 18:312–328CrossRefGoogle Scholar
  5. 5.
    Al-Taweel AM, Raish M, Perveen S et al (2017) Nepeta deflersiana attenuates isoproterenol-induced myocardial injuries in rats: possible involvement of oxidative stress, apoptosis, inflammation through nuclear factor (NF)-κB downregulation. Phytomedicine 34:67–75CrossRefGoogle Scholar
  6. 6.
    Suchal K, Malik S, Gamad N et al (2016) Mangiferin protect myocardial insults through modulation of MAPK/TGF-β pathways. Eur J Pharmacol 776:34–43CrossRefGoogle Scholar
  7. 7.
    Sun SJ, Wu XP, Song HL, Li GQ (2015) Baicalin ameliorates isoproterenol-induced acute myocardial infarction through iNOS, inflammation, oxidative stress and P38MAPK pathway in rat. Int J Clin Exp Med 8:22063–22072Google Scholar
  8. 8.
    Kocak C, Kocak FE, Akcilar R et al (2016) Molecular and biochemical evidence on the protective effects of embelin and carnosic acid in isoproterenol-induced acute myocardial injury in rats. Life Sci 147:15–23CrossRefGoogle Scholar
  9. 9.
    Chunhua M, Hongyan L, Weina Z et al (2017) Dang Gui Bu Xue Tang ameliorates coronary artery ligation-induced myocardial ischemia in rats. Biomed Pharmacother 88:617–624CrossRefGoogle Scholar
  10. 10.
    Naowaboot J, Wannasiri S, Pannangpetch P (2016) Morin attenuates hepatic insulin resistance in high-fat-diet-induced obese mice. J Physiol Biochem 72:269–280CrossRefGoogle Scholar
  11. 11.
    Kaltalioglu K, Coskun-Cevher S (2016) Potential of morin and hesperidin in the prevention of cisplatin-induced nephrotoxicity. Ren Fail 38:1291–1299CrossRefGoogle Scholar
  12. 12.
    Kuzu M, Kandemir FM, Yildirim S et al (2018) Morin attenuates doxorubicin-induced heart and brain damage by reducing oxidative stress, inflammation and apoptosis. Biomed Pharmacother 106:443–453CrossRefGoogle Scholar
  13. 13.
    Ben-Azu B, Aderibigbe AO, Eneni AO et al (2018) Morin attenuates neurochemical changes and increased oxidative/nitrergic stress in brains of mice exposed to ketamine: prevention and reversal of schizophrenia-like symptoms. Neurochem Res 43(9):1745–1755CrossRefGoogle Scholar
  14. 14.
    Wang N, Zhang J, Qin M et al (2018) Amelioration of streptozotocin-induced pancreatic β cell damage by morin: Involvement of the AMPK–FOXO3–catalase signaling pathway. Int J Mol Med 41(3):1409–1418Google Scholar
  15. 15.
    Lee KM, Lee Y, Chun HJ et al (2016) Neuroprotective and anti-inflammatory effects of morin in a murine model of Parkinson’s disease. J Neurosci Res 94:865–878CrossRefGoogle Scholar
  16. 16.
    Sinha K, Sadhukhan P, Saha S, Pal PB, Sil PC (2015) Morin protects gastric mucosa from nonsteroidal anti-inflammatory drug, indomethacin induced inflammatory damage and apoptosis by modulating NF-κB pathway. Biochim Biophys Acta 1850:769–783CrossRefGoogle Scholar
  17. 17.
    Tian Y, Li Z, Shen B, Zhang Q, Feng H (2017) Protective effects of morin on lipopolysaccharide/d-galactosamine-induced acute liver injury by inhibiting TLR4/NF-κB and activating Nrf2/HO-1 signalling pathways. Int Immunopharmacol 45:148–155CrossRefGoogle Scholar
  18. 18.
    Komirishetty P, Areti A, Sistla R, Kumar A (2016) Morin mitigates chronic constriction injury (CCI)-induced peripheral neuropathy by inhibiting oxidative stress induced PARP over activation and neuroinflammation. Neurochem Res 41:2029–2042CrossRefGoogle Scholar
  19. 19.
    Lee MH, Han MH, Lee DS et al (2017) Morin exerts cytoprotective effects against oxidative stress in C2C12 myoblasts via the upregulation of Nrf2-dependent HO-1 expression and the activation of the ERK pathway. Int J Mol Med 39:399–406CrossRefGoogle Scholar
  20. 20.
    Wei Z, He X, Kou J et al (2015) Renoprotective mechanisms of morin in cisplatin-induced kidney injury. Int Immunopharmacol 28:500–506CrossRefGoogle Scholar
  21. 21.
    Al-Numair KS, Chandramohan G, Alsaif MA (2012) Pretreatment with morin, a flavonoid, ameliorates adenosine triphosphatases and glycoproteins in isoproterenol-induced myocardial infarction in rats. J Nat Med 66:95–101CrossRefGoogle Scholar
  22. 22.
    Al-Numair KS, Chandramohan G, Alsaif MA et al (2014) Morin, a flavonoid, on lipid peroxidation and antioxidant status in experimental myocardial ischemic rats. Afr J Tradit Complement Altern Med 11:14–20CrossRefGoogle Scholar
  23. 23.
    Pogula BK, Maharajan MK, Oddepalli DR et al (2012) Morin protects heart from beta-adrenergic-stimulated myocardial infarction: an electrocardiographic, biochemical, and histological study in rats. J Physiol Biochem 68:433–446CrossRefGoogle Scholar
  24. 24.
    Al Numair KS, Chandramohan G, Alsaif MA, Baskar AA (2012) Protective effect of morin on cardiac mitochondrial function during isoproterenol-induced myocardial infarction in male Wistar rats. Redox Rep 17:14–21CrossRefGoogle Scholar
  25. 25.
    Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358CrossRefGoogle Scholar
  26. 26.
    Moron MS, Depierre JW, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 582:67–78CrossRefGoogle Scholar
  27. 27.
    Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474CrossRefGoogle Scholar
  28. 28.
    Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefGoogle Scholar
  29. 29.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  30. 30.
    Perez-Vizcaino F, Duarte J (2010) Flavonols and cardiovascular disease. Mol Aspects Med 31:478–494CrossRefGoogle Scholar
  31. 31.
    Ojha S, Taee HA, Goyal S et al (2016) Cardioprotective potentials of plant-derived small molecules against doxorubicin associated cardiotoxicity. Oxid Med Cell Longev. Google Scholar
  32. 32.
    Sachdeva J, Tanwar V, Golechha M et al (2012) Crocus sativus L. (saffron) attenuates isoproterenol-induced myocardial injury via preserving cardiac functions and strengthening antioxidant defense system. Exp Toxicol Pathol 64:557–564CrossRefGoogle Scholar
  33. 33.
    Hoffman JI, Buckberg GD (2014) The myocardial oxygen supply: demand index revisited. J Am Heart Assoc 3:e000285CrossRefGoogle Scholar
  34. 34.
    Stanley WC, Lopaschuk GD, Hall JL et al (1997) Regulation of myocardial carbohydrate metabolism under normal and ischaemic conditions: potential for pharmacological interventions. Cardiovasc Res 33:243–257CrossRefGoogle Scholar
  35. 35.
    Díaz-Muñoz M, Alvarez-Pérez MA, Yáñez L et al (2006) Correlation between oxidative stress and alteration of intracellular calcium handling in isoproterenol-induced myocardial infarction. Mol Cell Biochem 289:125–136CrossRefGoogle Scholar
  36. 36.
    Imahashi K, Pott C, Goldhaber JI et al (2005) Cardiac-specific ablation of the Na+–Ca2+ exchanger confers protection against ischemia/reperfusion injury. Circ Res 97:916–921CrossRefGoogle Scholar
  37. 37.
    Kaneko M, Matsumoto Y, Hayashi H et al (1994) Oxygen free radicals and calcium homeostasis in the heart. Mol Cell Biochem 139:91–100CrossRefGoogle Scholar
  38. 38.
    Rodrigo R, Libuy M, Feliu F, Hasson D (2013) Molecular basis of cardioprotective effect of antioxidant vitamins in myocardial infarction. Biomed Res Int. Google Scholar
  39. 39.
    Duilio C, Ambrosio G, Kuppusamy P et al (2001) Neutrophils are primary source of O2 radicals during reperfusion after prolonged myocardial ischemia. Am J Physiol Heart Circ Physiol 280:2649–2657CrossRefGoogle Scholar
  40. 40.
    Hori M, Nishida K (2009) Oxidative stress and left ventricular remodelling after myocardial infarction. Cardiovasc Res 81:457–464CrossRefGoogle Scholar
  41. 41.
    Khan V, Sharma S, Bhandari U et al (2018) Raspberry ketone protects against isoproterenol-induced myocardial infarction in rats. Life Sci 194:205–212CrossRefGoogle Scholar
  42. 42.
    Paul S, Das S, Tanvir EM et al (2017) Protective effects of ethanolic peel and pulp extracts of Citrus macroptera fruit against isoproterenol-induced myocardial infarction in rats. Biomed Pharmacother 94:256–264CrossRefGoogle Scholar
  43. 43.
    Chen Q, Jiang L, Li C et al (2012) Haemodynamics-driven developmental pruning of brain vasculature in zebrafish. PLoS Biol 10:e1001374CrossRefGoogle Scholar
  44. 44.
    Muslin AJ (2008) MAPK signalling in cardiovascular health and disease: molecular mechanisms and therapeutic targets. Clin Sci 115:203–218CrossRefGoogle Scholar
  45. 45.
    Dong LY, Li S, Zhen YL et al (2013) Cardioprotection of vitexin on myocardial ischemia/reperfusion injury in rat via regulating inflammatory cytokines and MAPK pathway. Am J Chin Med 41:1251–1266CrossRefGoogle Scholar
  46. 46.
    Khan SI, Malhotra RK, Rani N et al (2017) Febuxostat modulates MAPK/NF-κBp65/TNF-α signaling in cardiac ischemia-reperfusion injury. Oxid Med Cell Longev 2017:8095825Google Scholar
  47. 47.
    Kim JM, Lee EK, Park G et al (2010) Morin modulates the oxidative stress-induced NF-kappaB pathway through its anti-oxidant activity. Free Radic Res 44:454–461CrossRefGoogle Scholar
  48. 48.
    Wang J, Guo C, Wei Z et al (2016) Morin suppresses inflammatory cytokine expression by downregulation of nuclear factor-κB and mitogen-activated protein kinase (MAPK) signaling pathways in lipopolysaccharide-stimulated primary bovine mammary epithelial cells. J Dairy Sci 99:3016–3022CrossRefGoogle Scholar
  49. 49.
    Gupta SC, Tyagi AK, Deshmukh-Taskar P et al (2014) Down-regulation of tumor necrosis factor and other proinflammatory biomarkers by polyphenols. Arch Biochem Biophys 559:91–99CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Cardiovascular Research Laboratory, Department of PharmacologyAll India Institute of Medical SciencesNew DelhiIndia
  2. 2.Department of PharmacologyArmy College of Medical SciencesNew DelhiIndia

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