, Volume 70, Issue 2, pp 523–536 | Cite as

Spirulina platensis prevents high glucose-induced oxidative stress mitochondrial damage mediated apoptosis in cardiomyoblasts

  • Pratiksha Jadaun
  • Dhananjay Yadav
  • Prakash Singh BisenEmail author
Original Article


The current study was undertaken to study the effect of Spirulina platensis (Spirulina) extract on enhanced oxidative stress during high glucose induced cell death in H9c2 cells. H9c2 cultured under high glucose (33 mM) conditions resulted in a noteworthy increase in oxidative stress (free radical species) accompanied by loss of mitochondrial membrane potential, release of cytochrome c, increase in caspase activity and pro-apoptotic protein (Bax). Spirulina extract (1 μg/mL), considerably inhibited increased ROS and RNS levels, reduction in cytochrome c release, raise in mitochondrial membrane potential, decreased the over expression of proapoptotic protein Bax and suppressed the Bax/Bcl2 ratio with induced apoptosis without affecting cell viability. Overall results suggest that Spirulina extract plays preventing role against enhanced oxidative stress during high glucose induced apoptosis in cardiomyoblasts as well as related dysfunction in H9c2 cells.

Graphical Abstract


H9c2 High glucose Oxidative stress Apoptosis Spirulina platensis 



Normal glucose


High glucose


Spirulina platensis


Reactive oxygen species


Reactive nitrogen species




Hydrogen peroxide


Nitric oxide



X + XO

Xanthine plus xanthine oxidase






Copper–Zinc-superoxide dismutase


NG-nitro-l-arginine methyl ester



We thank, Director,  National Centre of Cell Sciences, NCCS Complex, S.P. Pune University, Ganeshkhind, Pune, Maharashtra, India for providing basic facilities and to Dr. Sandhya Sitasawad, Senior Scientist, National Centre of Cell Sciences, NCCS Complex, S.P. Pune University, Ganeshkhind, Pune, Maharashtra, India for her critical suggestion, support and encouragement. This research work was supported by a research grant from Department of Science and Technology, Govt. of India, New Delhi under FTYS scheme   (SR/FT/LS-093/2007).

Compliance with ethical standards

Conflict of interest

We declare that there is no conflict of interest.

Supplementary material

10616_2017_121_MOESM1_ESM.pptx (684 kb)
Supplementary material 1 (PPTX 684 kb)
10616_2017_121_MOESM2_ESM.docx (13 kb)
Supplementary material 2 (DOCX 12 kb)


  1. Abdel-Daim MM, Abuzead SM, Halawa SM (2013) Protective role of Spirulina platensis against acute deltamethrin-induced toxicity in rats. PLoS ONE 8:e72991CrossRefGoogle Scholar
  2. Allen DA, Harwood S, Varagunam M, Raftery MJ, Yaqoob MM (2003) High glucose-induced oxidative stress causes apoptosis in proximal tubular epithelial cells and is mediated by multiple caspases. FASEB J 17:908–910CrossRefGoogle Scholar
  3. Benedetti S, Benvenuti F, Pagilarani S, Francogli S, Scoglio S, Canestran F (2004) Antioxidant properties of a novel phycocyanin extract from the bluegreen alga Aphanizomenon flos-aquae. Life Sci 75:2353–2362CrossRefGoogle Scholar
  4. Brownlee M (2001) Biochemistry and molecular cell biology of diabetic complications. Nature 414:813–820CrossRefGoogle Scholar
  5. Budihardjo I, Oliver H, Lutter M, Luo X, Wang X (1999) Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol 15:269–290CrossRefGoogle Scholar
  6. Cai L, Kang YJ (2001) Oxidative stress and diabetic cardiomyopathy: a brief review. Cardiovasc Toxicol 1:181–193CrossRefGoogle Scholar
  7. Cai L, Li W, Wang G, Guo L, Jiang Y, Kang YJ (2002) Hyperglycemia-induced apoptosis in mouse myocardium: mitochondrial cytochrome C-mediated caspase-3 activation pathway. Diabetes 51:1938–1948CrossRefGoogle Scholar
  8. Carmody RJ, Cotter TG (2001) Signalling apoptosis: a radical approach. Redox Rep 6:77–90CrossRefGoogle Scholar
  9. Chandra J, Samali A, Orrenius S (2003) Triggering and modulation of apoptosis by oxidative stress. Free Radic Biol Med 29:323–333CrossRefGoogle Scholar
  10. Chen Y, Saari JT, Kang YJ (1995) Weak antioxidant defenses make the heart a target for damage in copper-deficient rats. Free Radic Biol Med 17:529–536CrossRefGoogle Scholar
  11. Chen T, Wong Y, Zheng W (2009) Induction of cell cycle arrest and mitochondria-mediated apoptosis in MCF-7 human breast carcinoma cells by selenium-enriched Spirulina extract. Biomed Pharmacother. doi: 10.1016/j.biopha.2009.09.006 Google Scholar
  12. Chen-Levy Z, Cleary ML (1990) Membrane topology of the Bcl-2 proto-oncogenic protein demonstrated in vitro. Biol Chem 265:4929–4933Google Scholar
  13. Chu W-L, Lim Y-W, Radhakrishnan AK, Lim P-E (2010) Protective effect of aqueous extract from Spirulina platensis against cell death induced by free radicals. BMC Complement Altern Med 10:53.
  14. Cooke MS, Evans MD, Dizdaroglu M, Lunec J (2003) Oxidative DNA damage: mechanisms, mutation, and disease. FASEB J 17:1195–1214CrossRefGoogle Scholar
  15. Desagher S, Martinou JC (2000) Mitochondria as the central control point of apoptosis. Trends Cell Biol 10:369–377CrossRefGoogle Scholar
  16. Felix C, Gillis M, Driedzic WR, Paulson DJ, Broderick TL (2001) Effects of propionyl-l-carnitine on isolated mitochondrial function in the reperfused diabetic rat heart. Diabetes Res Clin Pract 53:17–24CrossRefGoogle Scholar
  17. Gupta R, Bhadauriya P, Chauhan VS, Bisen PS (2008) Impact of UV-B radiation on thylakoid membrane and fatty acid profile of Spirulina platensis. Curr Microbiol 56:156–161CrossRefGoogle Scholar
  18. Gustafsson AB, Gottlieb RA (2007) Bcl-2 family members and apoptosis, taken to heart. Am J Physiol Cell Physiol 292:C45–C51CrossRefGoogle Scholar
  19. Hand SC, Menze MA (2008) Mitochondria in energy-limited states: mechanisms that blunt the signaling of cell death. J Exp Biol 211:1829–1840CrossRefGoogle Scholar
  20. Hwang JH, Chen JC, Chan YC (2013) Effects of C-phycocyanin and Spirulina on salicylate-induced tinnitus, expression of NMDA receptor and inflammatory genes. PLoS ONE 8:e58215CrossRefGoogle Scholar
  21. Jin HJ, Xie XL, Ye JMC-G, Li CG (2013) TanshinoneIIA and cryptotanshinone protect against hypoxia-induced mitochondrial apoptosis in H9c2 Cells. PLoS ONE 8:e51720. doi: 10.1371/journal.pone.0051720 CrossRefGoogle Scholar
  22. Kaul N, Siveski Iliskovic N, Hill M, Khaper N, Seneviratne C, Singal PK (1996) Probucol treatment reverses antioxidant and functional deficit in diabetic cardiomyopathy. Mol Cell Biochem 160:283–288CrossRefGoogle Scholar
  23. Khan Z, Bhadouria P, Bisen PS (2005) Nutritional and therapeutic potential of Spirulina. Curr Pharm Biotechnol 6:373–379CrossRefGoogle Scholar
  24. Kinoshita T, Yokota T, Arai K, Miyajima A (1995) Regulation of Bcl2 expression of oncogenic Ras protein in hematopoietic cells. Oncogene 10:2207–2212Google Scholar
  25. Kulshreshtha A, Zacharia AJ, Jarouliya U, Bhadauriya P, Prasad GBKS, Bisen PS (2008) Spirulina in health care management. Curr Pharm Biotechnol 9:400–405CrossRefGoogle Scholar
  26. Kumar S, Sitasawad SL (2009) N-acetylcysteine prevents glucose/glucose oxidase-induced oxidative stress, mitochondrial damage and apoptosis in H9c2 cells. Life Sci 84:328–336CrossRefGoogle Scholar
  27. Kumar S, Kain V, Sitasawad SL (2012) High glucose-induced Ca2+ overload and oxidative stress contribute to apoptosis of cardiac cells through mitochondrial dependent and independent pathways. Biochim Biophys Acta 1820:907–920CrossRefGoogle Scholar
  28. Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91:92–104CrossRefGoogle Scholar
  29. Li XL, Xu G, Chen T, Wong YS, Zhao HL, Fan RR, Gu XM, Tong PC, Chan JC (2009) Phycocyanin protects INS-1E pancreatic beta cells against human islet amyloid polypeptide-induced apoptosis through attenuating oxidative stress and modulating JNK and p38 mitogen-activated protein kinase pathways. Int J Biochem Cell Biol 41:1526–1535CrossRefGoogle Scholar
  30. Mates JM (2000) Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology. Toxicology 153:83–104CrossRefGoogle Scholar
  31. Mayer B, Oberbauer R (2003) Mitochondrial regulation of apoptosis. Physiology 18:89–94CrossRefGoogle Scholar
  32. Nishikawa T, Edelstein D, Du XL, Yamagishi S, Matsumura T, Kaneda Y, Yorek MA, Beebe D, Oates PJ, Hammes HP, Giardino I, Brownlee M (2000) Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature 404:787–790CrossRefGoogle Scholar
  33. Oltvai ZN, Milliman CL, Korsmeyer SI (1993) Bcl-2 heterodimerizes in vitro with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74:609–619CrossRefGoogle Scholar
  34. Romay C, Ledon N, Gonzalez R (1999) Phycocyanin extract reduces leukotriene B4 levels in arachidonic acid-induced mouse-ear inflammation test. J Pharm Pharmacol 51:641–642CrossRefGoogle Scholar
  35. Rosen P, Ballhausen T, BlochW Addicks K (1995) Endothelial relaxation is disturbed by oxidative stress in the diabetic rat heart: influence of tocopherol as antioxidant. Diabetologia 38:1157–1168CrossRefGoogle Scholar
  36. Samuels R, Mani UV, Iyer UM, Nayak US (2002) Hypocholesterolemic effect of Spirulina in patients with hyperlipidemic nephrotic syndrome. J Med Food 5:91–96CrossRefGoogle Scholar
  37. Sanghvi AM, Lo YM (2010) Present and potential industrial applications of macro- and microalgae. Recent Pat Food Nutr Agric 2:187–194CrossRefGoogle Scholar
  38. Shih CM, Ko WC, Wu JS, Wei YH, Wang LF, Chang EE, Lo TY, Cheng HH, Chen CT (2004) Mediating of caspase-independent apoptosis by cadmium through the mitochondria-ROS pathway in MRC-5 fibroblasts. J Cell Biochem 91:384–397CrossRefGoogle Scholar
  39. Silveira ST, Burkert JF, Costa JA, Burkert CA, Kalil SJ (2007) Optimization of phycocyanin extraction from Spirulina platensis using factorial design. Bioresour Technol 98:1629–1634CrossRefGoogle Scholar
  40. Smogorzewski M, Galfayan V, Massry SG (1998) High glucose concentration causes a rise in [Ca2+]i of cardiac myocytes. Kidney Int 53:1237–1243CrossRefGoogle Scholar
  41. Susin SA, Zamzami N, Castedo M, Hirsch T, Marchetti P, Macho A, Daugas E, Gauskens M, Kroemer G (1996) Bcl-2 inhibits the mitochondrial release of an apoptogenic protease. J Exp Med 184:1331–1341CrossRefGoogle Scholar
  42. Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, Prévost MC, Alzari PM, Kroemer G (1999) Mitochondrial release of caspase-2 and -9 during the apoptotic process. J Exp Med 189:381–394CrossRefGoogle Scholar
  43. Tsai KH, Wang WJ, Lin CW, Pai P, Lai TY, Tsai CY, Kuo WW (2012) NADPH oxidase-derived superoxide anion-induced apoptosis is mediated via the JNK-dependent activation of NF-kappaB in cardiomyocytes exposed to high glucose. J Cell Physiol 227:1347–1357CrossRefGoogle Scholar
  44. Verzola D, Bertolotto MB, Villaggio B, Ottonello L, Dallegri F, Salvatore F, Berruti V, Gandolfo MT, Garibotto G, Deferrari G (2004) Oxidative stress mediates apoptotic changes induced by hyperglycemia in human tubular kidney cells. J Am Soc Nephrol 15:S85–S87CrossRefGoogle Scholar
  45. Wu Q, Liu L, Miron A, Klímová B, Wan D, Kuc K (2016) The antioxidant, immunomodulatory, and anti-inflammatory activities of Spirulina: an overview. Toxicol, Arch. doi: 10.1007/s00204-016-1744-5 Google Scholar
  46. Yang L, Mashima T, Sato S, Mochizuki M, Sakamoto H, Yamori T, Oh-Hara T, Tsuruo T (2003) Predominant suppression of apoptosome by inhibitor of apoptosis protein in non-small cell lung cancer. H460 cells: therapeutic affects of a novel polyarginine-conjugated Smac peptide. Cancer Res 63:831–837Google Scholar
  47. Yin XM, Oltvai ZN, Korsmeyer SJ (1995) Heterodimerization with Bax is required for Bcl-2 to repress cell death. Curr Trop Microbiol Immunol 194:331–338Google Scholar
  48. Yu T, Sheu SS, Robotham JL, Yoon Y (2008) Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species. Cardiovasc Res 79:341–351CrossRefGoogle Scholar
  49. Zheng J, Inoguch T, Sasaki S, Maeda Y, McCarty MF, Fujii M, Ikeda N, Kobayashi K, Sonoda N, Takayanagi R (2013) Phycocyanin and phycocyanobilin from Spirulina platensis protect against diabetic nephropathy by inhibiting oxidative stress. Am J Physiol Regul Integr Comp Physiol 304:R110–R120CrossRefGoogle Scholar
  50. Zhou ZP, Liu LN, Chen XL, Wang JX, Chen M, Zhang YZ, Zhou BC (2005) Factors that affect antioxidant activity of c-phycocyanins from Spirulina platensis. J Food Biochem 29:313–322CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Pratiksha Jadaun
    • 1
  • Dhananjay Yadav
    • 2
  • Prakash Singh Bisen
    • 3
    Email author
  1. 1.School of Studies in BiotechnologyGwaliorIndia
  2. 2.Department of Medical BiotechnologyYeungnam UniversityGyeongsanKorea
  3. 3.School of Studies in BiotechnologyJiwaji UniversityGwaliorIndia

Personalised recommendations