Journal of Physiology and Biochemistry

, Volume 71, Issue 2, pp 289–299 | Cite as

Vitamin D attenuates pro-inflammatory TNF-α cytokine expression by inhibiting NF-кB/p65 signaling in hypertrophied rat hearts

  • Nawal M. Al-Rasheed
  • Nouf M. Al-Rasheed
  • Yieldez A. Bassiouni
  • Iman H. HasanEmail author
  • Maha A. Al-Amin
  • Hanaa N. Al-Ajmi
  • Raeesa A. Mohamad
Original Paper


A growing body of evidence suggests that immune activation and inflammatory mediators may play a key role in the development and progression of left ventricle (LV) hypertrophy. The present study was designed to test the hypothesis that the cardioprotective effect of cholecalciferol (Vit-D3) is mediated via the regulation of messenger RNA (mRNA) expression of pro-inflammatory cytokines. Rats were randomly divided into four groups: control group received normal saline (0.9 % NaCl) i.p. for 14 days; Vit-D3 group received Vit-D3 at a dose of 12 μg/kg/day by gavage for 14 days; ISO group received saline for 7 days, and at day 7, ISO (5 mg/kg/day) was injected i.p. for 7 consecutive days to induce cardiac hypertrophy; and Vit-D3 + ISO group was treated with Vit-D3 for 14 days, and at day 7, ISO was administered for 7 consecutive days. Heart/body weight ratio, troponin-T, creatine kinase-MB, and tumor necrosis factor-α (TNF-α) levels of LV tissue were estimated. Levels of mRNA expression of NF-кB (NF-кB)/p65 and inhibitory kappa B (IкB)-α were determined by real-time PCR. Vit-D3 administration before and during induction of cardiac hypertrophy significantly reduced (P < 0.001) cardiac biomarkers. The histopathological examination further confirmed these results. In addition, Vit-D3 significantly decreased (P < 0.001) NF-кB-p65 mRNA expression and increased (P < 0.01) IкB-α mRNA expression in LV tissues compared to ISO group. Based on these findings, it was concluded that the administration of cholecalciferol markedly attenuated the development of ISO-induced cardiac hypertrophy likely through downregulation of TNF-α /NF-кb/p65 signaling pathways. However, it should be pointed out that other signaling pathways may contribute to the cardioprotective effect of Vit-D3 which requires further investigation.


Cholecalciferol mRNA NF-кB-p65/IкB-α signaling Cardiac hypertrophy Rats 



This research project was supported by a grant from the “Research Center of the Female Scientific and Medical Colleges”, Deanship of Scientific Research, King Saud University.

Conflict of interest

The authors declare that they have no conflict of interest.

Authors’ contribution

NMA and NA participated in the experimental design and revised the final manuscript. YA wrote and revised the all results of the manuscript. IH carried out the molecular genetic studies, performed the statistical analysis, and participated in the sequence alignment and drafted the manuscript. MA and HN designed the experimental and participated in the biochemical analysis and data collections. RA carried out the histological and immunoassays. All authors read and approved the final manuscript.


  1. 1.
    Adams LS, Teegarden D (2004) 1,25-Dihydroxycholecalciferol inhibits apoptosis in C3H10T1/2 murine fibroblast cells through activation of nuclear factor kappaB. J Nutr 134:2948–2952PubMedGoogle Scholar
  2. 2.
    Artaza JN, Rajnish M, Norris KC (2009) Vitamin D and the cardiovascular system. Clin J Am Soc Nephrol 4:1515–1522CrossRefPubMedGoogle Scholar
  3. 3.
    Bonizzi G, Karin M (2004) The two NF-kappaB activation pathways and their role in innate and adaptive immunity. Trends Immunol 25:280–288CrossRefPubMedGoogle Scholar
  4. 4.
    Bujak M, Ren G, Kweon HJ, Dobaczewski M, Reddy A, Taffet G, Wang XF, Frangogiannis NG (2007) Essential role of Smad3 in infarct healing and in the pathogenesis of cardiac remodeling. Circulation 116:2127–2138CrossRefPubMedGoogle Scholar
  5. 5.
    Choi M, Park H, Cho S, Lee M (2013) Vitamin D3 supplementation modulates inflammatory responses from the muscle damage induced by high-intensity exercise in SD rats. Cytokine 63:27–35CrossRefPubMedGoogle Scholar
  6. 6.
    Cohen-Lahav M, Shany S, Tobvin D, Chaimovitz C, Douvdevani A (2006) Vitamin D decreases NFkappaB activity by increasing IkappaBalpha levels. Nephrol Dial Transplant 21:889–897CrossRefPubMedGoogle Scholar
  7. 7.
    Condorelli G, Morisco C, Latronico MV (2002) TNF alpha signal transductionin rat neonatal cardiacmyocytes: definition of pathways generating from the TNF-alpha receptor. FASEBJ 16:1732–1737CrossRefGoogle Scholar
  8. 8.
    Fleet JC (2008) Molecular actions of vitamin D contributing to cancer prevention. Mol Asp Med 29:388–396CrossRefGoogle Scholar
  9. 9.
    Gong KZ, Song G, Spiers JP, Kelso EJ, Zhang ZG (2007) Activation of immune and inflammatory systems in chronic heart failure: novel therapeutic approaches. Int J Clin Pract 61(4):611–621CrossRefPubMedGoogle Scholar
  10. 10.
    Gupta K, Agrawal T, DelCore MG, Mohiuddin SM, Agrawal DK (2012) Vitamin D deficiency induces cardiac hypertrophy and inflammation in epicardial adipose tissue in hypercholesterolemic swine. Exp Mol Pathol 93:82–90CrossRefPubMedCentralPubMedGoogle Scholar
  11. 11.
    Hadi AM, Mouchaers KTB, Schalij I, Grunberg K, Meijer GA, Vonk-Noordegraaf A, van der Laarse WJ, Beliën JAM (2011) Rapid quantification of myocardial fibrosis: a new macro-based automated analysis. Cell Oncol (Dordr) 34(4):343–354CrossRefGoogle Scholar
  12. 12.
    Hamid T, Guo SZ, Kingery JR, Xiang X, Dawn B, Prabhu SD (2011) Cardiomyocyte NF-кB p65 promotes adverse remodelling, apoptosis, and endoplasmic reticulum stress in heart failure. Cardiovasc Res 89(1):129–138CrossRefPubMedCentralPubMedGoogle Scholar
  13. 13.
    Hansi Priscilla D, Stanely Mainzen Prince P (2009) Cardioprotective effect of gallic acid on cardiac troponin-T, cardiac marker enzymes, lipid peroxidation products and antioxidants in experimentally induced myocardial infarction in Wistar rats. Chem Biol Interact 179:118–124CrossRefPubMedGoogle Scholar
  14. 14.
    Harant H, Andrew PJ, Reddy GS, Foglar E, Lindley IJ (1997) 1 Alpha,25-dihydroxyvitamin D3 and a variety of its natural metabolites transcriptionally repress nuclear-factor-kappaB-mediated interleukin-8 gene expression. Eur J Biochem 250:63–71CrossRefPubMedGoogle Scholar
  15. 15.
    Hingtgen SD, Li Z, Kutschke W, Tian X, Sharma RV, Davisson RL (2010) Superoxide scavenging and Akt inhibition in myocardium ameliorate pressure overload-induced NF-jB activation and cardiac hypertrophy. Physiol Genomics 41(2):127–136CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    Jobe LJ, Melendez GC, Levick SP, Du Y, Brower GL, Janicki JS (2009) TNF-alpha inhibition attenuates adverse myocardial remodeling in a rat model of volume overload. Am J Physiol Heart Circ Physiol 297:1462–1468CrossRefGoogle Scholar
  17. 17.
    Jobe LJ, Meléndez GC, Levick SP, Du Y, Brower GL, Janicki JS (2009) Am J Physiol Heart Circ Physiol 297:4CrossRefGoogle Scholar
  18. 18.
    Kapadia SR, Oral H, Lee J, Nakano M, Taffet GE, Mann DL (1997) Hemodynamic regulation of tumor necrosis factor-alpha gene and protein expression in adult feline myocardium. Circ Res 81:187–195CrossRefPubMedGoogle Scholar
  19. 19.
    Kenneth J, Livak, Thomas D (2001) Schmittgen analysis of relative gene expression data using real-time quantitative PCR and the 22DDCT. Methods 25:402–408CrossRefGoogle Scholar
  20. 20.
    Levine B, Kalman J, Mayer L, Fillit H, Packer M (1990) Elevated circulating levels of tumor necrosis factor in severe chronic heart failure. N Engl J Med 323(4):236–241CrossRefPubMedGoogle Scholar
  21. 21.
    Leychenko A, Konorev E, Jijiwa M, Matter ML (2011) Stretch-induced hypertrophy activates NFkB-mediated VEGF secretion in adult cardiomyocytes. PLoS One 6(12):e29055CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Li N, Karin M (1999) Is NF-кB the sensor of oxidative stress? FASEB J 13:1137–1143PubMedGoogle Scholar
  23. 23.
    Meems LM, van der Harst P, van Gilst WH, de Boer RA (2011) Vitamin D biology in heart failure: molecular mechanisms and systematic review. Curr Drug Targets 12:29–41CrossRefPubMedGoogle Scholar
  24. 24.
    Mercurio F, Manning AM (1999) NF-кB as a primary regulator of the stress response. Oncogene 18:6163–6171CrossRefPubMedGoogle Scholar
  25. 25.
    Mészáros J, Khananshvili D, Hart G (2001) Mechanisms underlying delayed after depolarizations in hypertrophied left ventricular myocytes of rats. Am J Phys Heart Circ Physiol 281:903–914Google Scholar
  26. 26.
    Muller K, Haahr P, Diamant M, Rieneck K, Kharazmi A, Bendtzen K (1992) 1,25-Dihydroxyvitamin D3 inhibits cytokine production by human blood monocytes at the post-transcriptional level. Cytokine 4:506–512CrossRefPubMedGoogle Scholar
  27. 27.
    Mustapha S, Kirshner A, De Moissac D, Kirshenbaum LA (2000) A direct requirement of nuclear factor-kappa B for suppression of apoptosis in ventricular myocytes. Am J Physiol Heart Circ Physiol 279(3):H939–H945PubMedGoogle Scholar
  28. 28.
    Nagpal S, Na S, Rathnachalam R (2005) Noncalcemic actions of vitamin D receptor ligands. Endocr Rev 26:662–687CrossRefPubMedGoogle Scholar
  29. 29.
    Nagueh SF, Stetson SJ, Lakkis NM, Killip D, Perez-Verdia A, Entman ML, Spencer WH III, Torre-Amione G (2001) Decreased expression of tumor necrosis factor-alpha and regression of hypertrophy after nonsurgical septal reduction therapy for patients with hypertrophic obstructive cardiomyopathy. Circulation 103:1844–1850CrossRefPubMedGoogle Scholar
  30. 30.
    Palomer X, Alvarez-Guardia D, Rodríguez-Calvo R, Coll T, Laguna JC, Davidson MM, Chan TO, Feldman AM, Vázquez-Carrera M (2009) TNF-alpha reduces PGC-1alpha expression through NF-kappaB and p38 MAPK leading to increased glucose oxidation in a human cardiac cell model. Cardiovasc Res 81(4):703–712CrossRefPubMedGoogle Scholar
  31. 31.
    Riis JL, Johansen C, Gesser B, Møller K, Larsen CG, Kragballe K, Iversen L (2004) 1Alpha,25(OH)(2)D(3) regulates NF-kappaB DNA binding activity in cultured normal human keratinocytes through an increase in IkappaBalpha expression. Arch Dermatol Res 296:195–202CrossRefPubMedGoogle Scholar
  32. 32.
    Schuster M, Annemann M, Plaza-Sirvent C, Schmitz I (2013) Atypical IκB proteins—nuclear modulators of NF-κB signaling. Cell Commun Sign 11:23CrossRefGoogle Scholar
  33. 33.
    Sorriento D, Santulli G, Fusco A, Anastasio A, Trimarco B, Iaccarino G (2010) Intracardiac injection of AdGRK5-NT reduces left ventricular hypertrophy by inhibiting NFkappaB-dependent hypertrophic gene expression. Hypertension 56(4):696–704CrossRefPubMedGoogle Scholar
  34. 34.
    Sun J, Mustafi R, Cerda S, Chumsangsri A, Xia YR, Li YC, Bissonnette M (2008) Lithocholic acid down-regulation of NF-kappaB activity through vitamin D receptor in colonic cancer cells. J Steroid Biochem Mol Biol 111:37–40CrossRefPubMedCentralPubMedGoogle Scholar
  35. 35.
    Szeto FL, Sun J, Kong J, Duan Y, Liao A, Madara JL, Li YC (2007) Involvement of the vitamin D receptor in the regulation of NF-_B activity in fibroblasts. J Steroid Biochem Mol Biol 103:563–566CrossRefPubMedCentralPubMedGoogle Scholar
  36. 36.
    Tse AK, Wan CK, Shen XL, Zhu GY, Cheung HY, Yang M, Fong WF (2007) 1,25-Dihydroxyvitamin D3 induces biphasic NF-kappaB responses during HL-60 leukemia cells differentiation through protein induction and PI3K/Akt-dependent phosphorylation/degradation of IkappaB. Exp Cell Res 313:1722–1734CrossRefPubMedGoogle Scholar
  37. 37.
    Van Antwarp DJ, Martin SJ, Kafrt T, Green DR, Verma IM (1996) Suppression of TNF-α-induced apoptosis by NF-_B. Science 274:787–789CrossRefGoogle Scholar
  38. 38.
    Wang CY, Mayo MW, Waldwin AS Jr (1996) TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-кB. Science 274:784–787CrossRefPubMedGoogle Scholar
  39. 39.
    Weishaar RE, Kim SN, Saunders DE, Simpson RE (1990) Involvement of vitamin D3 with cardiovascular function III. Effect on physical and morphological properties. Am J Physiol 258:134–142Google Scholar
  40. 40.
    Yang L, Wang J, Fan Y, Chen S, Wang L, Ma J (2011) Effect of 1,25 (OH) 2D3 on rat peritoneal mesothelial cells treated with high glucose plus lipopolysaccharide. Cell Immunol 271:173–179CrossRefPubMedGoogle Scholar
  41. 41.
    Yu XW, Chen Q, Kennedy RH (2005) Inhibition of sarcoplasm micreticular function by chronic interleukin-6 exposure via NOS in adult ventricular myocytes. J Physiol 566:327–340CrossRefPubMedCentralPubMedGoogle Scholar
  42. 42.
    Zhang Z, Yuan W, Sun L, Szeto FL, Wong KE, Li X, Kong J, Li YC (2007) 1,25-Dihydroxyvitamin D3 targeting of NF-kappaB suppresses high glucose-induced MCP-1 expression in mesangial cells. Kidney Int 72:193–201CrossRefPubMedGoogle Scholar
  43. 43.
    Zhang T, Yang S, Du J (2014) Protective effects of berberine on isoproterenol-induced acute myocardial ischemia in rats through regulating HMGB1-TLR4 Axis. Evidence-Based Complementary and Alternative Medicine: Article ID 849783, 8 pagesGoogle Scholar
  44. 44.
    Zordoky BN, Aboutabl ME, El-Kadi AO (2008) Modulation of cytochrome P450 gene expression and arachidonic acid metabolism during isoproterenol-induced cardiac hypertrophy in rats. Drug Metab Dispos 36:2277–2286CrossRefPubMedGoogle Scholar

Copyright information

© University of Navarra 2015

Authors and Affiliations

  • Nawal M. Al-Rasheed
    • 1
  • Nouf M. Al-Rasheed
    • 1
  • Yieldez A. Bassiouni
    • 1
    • 2
  • Iman H. Hasan
    • 1
    • 4
    Email author
  • Maha A. Al-Amin
    • 1
  • Hanaa N. Al-Ajmi
    • 1
  • Raeesa A. Mohamad
    • 3
  1. 1.Pharmacology Department, Faculty of PharmacyKing Saud UniversityRiyadhKingdom of Saudi Arabia
  2. 2.Department of Pharmacology, Faculty of MedicineAlexandria UniversityAlexandriaEgypt
  3. 3.Department of Anatomy, College of MedicineKing Saud UniversityRiyadhSaudi Arabia
  4. 4.Department of Pharmacology and Toxicology, Faculty of PharmacyKing Saud UniversityRiyadhSaudi Arabia

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