Indole-3-carbinol improves neurobehavioral symptoms in a cerebral ischemic stroke model

  • Pankaj Paliwal
  • Gaurav Chauhan
  • Deepa Gautam
  • Debabrata Dash
  • Shashikant C. U. Patne
  • Sairam Krishnamurthy
Original Article

Abstract

Stroke is one of the most common causes of death worldwide and also responsible for permanent disability. Ischemic stroke has been found to affect 80% of stroke patients. Recombinant tissue plasminogen activator (rtPA) is the widely used drug for the ischemic stroke with narrow therapeutic window. Indole-3-carbinol (I3C) is a natural compound obtained from brassica species having antithrombotic activity. Middle cerebral artery occlusion (MCAO) model was used followed by reperfusion after 2 h of ischemia for the evaluation of the I3C against ischemic stroke. After reperfusion, I3C (12.5, 25, and 50 mg/kg) was given by oral route once daily and continued up to the 14th day. Behavioral studies including postural reflex, forelimb placing, and cylinder tests showed I3C attenuated the MCAO-induced increase in average score and asymmetry score efficiently. Mean cerebral blood flow (CBF) was improved by treatment with I3C (12.5 mg/kg) by 60% of baseline at 6 h. I3C inhibited ADP-induced platelet aggregation and reduced ischemic volume significantly. It also inhibited in vitro the ADP-induced platelet aggregation in healthy human volunteers. I3C improves behavioral scores and mean CBF after focal cerebral ischemia in rats. Furthermore, I3C showed prophylactic anti-thrombotic activity against carrageenan induced tail thrombosis. Therefore, preclinical evidence points to I3C as a potential candidate for use in cerebral ischemic stroke.

Keywords

Indole-3-carbinol Cerebral ischemia Middle cerebral artery occlusion Antithrombotic Cerebral blood flow 

Notes

Acknowledgements

None.

Compliance with ethical standards

Conflict of interest

P.P, G.C and S.K are inventors of an Indian patent application “An Oral Composition of Indole-3-Carbinol and A Method of Preparation Thereof” (No. 201711026941, 28/July/2017). All other authors declare no conflict of interest.

Research involving human participants and animals

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Institutional Ethics Committee, Institute of Medical Sciences, Banaras Hindu University and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. All procedures performed in studies involving animals were in accordance with the ethical standards of the Institutional Animal Ethics Committee at which the studies were conducted (Protocol no. Dean/2016/CAEC/31).

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. Abumiya T, Fitridge R, Mazur C, Copeland BR, Koziol JA, Tschopp JF, Pierschbacher MD, Del Zoppo GJ (2000) Integrin αIIbβ3 inhibitor preserves microvascular patency in experimental acute focal cerebral ischemia. Stroke 31(6):1402–1410.  https://doi.org/10.1161/01.STR.31.6.1402 CrossRefPubMedGoogle Scholar
  2. Aggarwal BB, Shishodia S (2006) Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharmacol 71:1397–1421.  https://doi.org/10.1016/j.bcp.2006.02.009 CrossRefPubMedGoogle Scholar
  3. Ampofo E, Lachnitt N, Rudzitis-Auth J, Schmitt BM, Menger MD, Laschke MW (2017) Indole-3-carbinol is a potent inhibitor of ischemia–reperfusion–induced inflammation. J Surg Res 215:34–46.  https://doi.org/10.1016/j.jss.2017.03.019 CrossRefPubMedGoogle Scholar
  4. Anderton MJ, Manson MM, Verschoyle RD, Gescher A, Lamb JH, Farmer PB, Steward WP, Williams ML (2004) Pharmacokinetics and tissue disposition of indole-3-carbinol and its acid condensation products after oral administration to mice. Clin Cancer Res 10(15):5233–5241.  https://doi.org/10.1158/1078-0432.CCR-04-0163 CrossRefPubMedGoogle Scholar
  5. Atlan M, Forget BC, Boccara AC, Vitalis T, Rancillac A, Dunn AK, Gross M (2007) Cortical blood flow assessment with frequency-domain laser Doppler microscopy. J Biomed Opt 12:024019.  https://doi.org/10.1117/1.2715184 CrossRefPubMedGoogle Scholar
  6. Ban JY, Kang SW, Lee JS, Chung JH, Ko YG, Choi HS (2012) Korean red ginseng protects against neuronal damage induced by transient focal ischemia in rats. Exp Ther Med 3:693–698.  https://doi.org/10.3892/etm.2012.449 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Bederson JB, Pitts LH, Tsuji M, Nishimura M, Davis R, Bartkowski H (1986) Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke 17:472–476.  https://doi.org/10.1161/01.STR.17.3.472 CrossRefPubMedGoogle Scholar
  8. Belayev L, Alonso OF, Busto R, Zhao W, Ginsberg MD (1996) Middle cerebral artery occlusion in the rat by intraluminal suture neurological and pathological evaluation of an improved model. Stroke 27:1616–1623.  https://doi.org/10.1161/01.STR.27.9.1616 CrossRefPubMedGoogle Scholar
  9. Belayev L, Khoutorova L, Deisher TA, Belayev A, Busto R, Zhang Y, Zhao W, Ginsberg MD (2003) Neuroprotective effect of SolCD39, a novel platelet aggregation inhibitor, on transient middle cerebral artery occlusion in rats. Stroke 34(3):758–763.  https://doi.org/10.1161/01.STR.0000056169.45365.15 CrossRefPubMedGoogle Scholar
  10. Berti R, Williams AJ, Moffett JR, Hale SL, Velarde LC, Elliott PJ, Yao C, Dave JR, Tortella FC (2002) Quantitative real-time RT-PCR analysis of inflammatory gene expression associated with ischemia-reperfusion brain injury. J Cereb Blood Flow 22(9):1068–1079.  https://doi.org/10.1097/00004647-200209000-00004 CrossRefGoogle Scholar
  11. Born GV, Wehmeier A (1979) Inhibition of platelet thrombus formation by chloropromazine acting to diminish haemolysis. Nature 282:212–213.  https://doi.org/10.1038/282212a0 CrossRefPubMedGoogle Scholar
  12. Cha JK, Jeon HW, Kang MJ (2008) ADP induced platelet aggregation in acute ischemic stroke patients on aspirin therapy. Eur J Neurol 15(12):1304–1308.  https://doi.org/10.1111/j.1468-1331.2008.02306.x CrossRefPubMedGoogle Scholar
  13. Choudhri TF, Hoh BL, Zerwes HG, Prestigiacomo CJ, Kim SC, Connolly ES, Kottirsch G, Pinsky DJ (1998) Reduced microvascular thrombosis and improved outcome in acute murine stroke by inhibiting GP IIb/IIIa receptor-mediated platelet aggregation. J Clin Invest 102(7):1301–1310.  https://doi.org/10.1172/JCI3338 CrossRefPubMedPubMedCentralGoogle Scholar
  14. Cui K, Luo X, Xu K, Murthy MV (2004) Role of oxidative stress in neurodegeneration: recent developments in assay methods for oxidative stress and nutraceutical antioxidants. Prog Neuro-Psychopharmacol Biol Psychiatry 28:771–799.  https://doi.org/10.1016/j.pnpbp.2004.05.023 CrossRefGoogle Scholar
  15. De Ryck M, Van Reempts J, Borgers M, Wauquier A, Janssen PA (1989) Photochemical stroke model: flunarizine prevents sensorimotor deficits after neocortical infarcts in rats. Stroke 20:1383–1390.  https://doi.org/10.1161/01.STR.20.10.1383 CrossRefPubMedGoogle Scholar
  16. Deb P, Sharma S, Hassan K (2010) Pathophysiologic mechanisms of acute ischemic stroke: an overview with emphasis on therapeutic significance beyond thrombolysis. Pathophysiology 17:197–218.  https://doi.org/10.1016/j.pathophys.2009.12.001 CrossRefPubMedGoogle Scholar
  17. Dhurat R, Sukesh MS (2014) Principles and methods of preparation of platelet-rich plasma: a review and author’s perspective. J Cutan Aesthet Surg 7(4):189–197.  https://doi.org/10.4103/0974-2077.150734 CrossRefPubMedPubMedCentralGoogle Scholar
  18. El-Naga RN, Ahmed HI, Al Haleem EN (2014) Effects of indole-3-carbinol on clonidine-induced neurotoxicity in rats: impact on oxidative stress, inflammation, apoptosis and monoamine levels. Neurotoxicology 44:48–57.  https://doi.org/10.1016/j.neuro.2014.05.004 CrossRefPubMedGoogle Scholar
  19. Ezzati M, Vander Hoorn S, Rodgers A, Lopez AD, Mathers CD, Murray CJ (2003) Estimates of global and regional potential health gains from reducing muliple major risk factors. Lancet 362:271–280.  https://doi.org/10.1016/S0140-6736(03)13968-2 CrossRefPubMedGoogle Scholar
  20. Fahlenkamp AV, Coburn M, de Prada A, Gereitzig N, Beyer C, Haase H, Rossaint R, Gempt J, Ryang YM (2014) Expression analysis following argon treatment in an in vivo model of transient middle cerebral artery occlusion in rats. Med Gas Res 4:11.  https://doi.org/10.1186/2045-9912-4-11 CrossRefPubMedPubMedCentralGoogle Scholar
  21. Fisher M (2004) The ischemic penumbra: identification, evolution and treatment concepts. Cerebrovasc Dis 17:1–6.  https://doi.org/10.1159/000074790 CrossRefPubMedGoogle Scholar
  22. Freedman JE (2008) Oxidative stress and platelets. Arterioscler Thromb Vasc Biol 28(3):11–16.  https://doi.org/10.1161/ATVBAHA.107.159178 CrossRefGoogle Scholar
  23. Guo Q, Wang G, Namura S (2010) Fenofibrate improves cerebral blood flow after middle cerebral artery occlusion in mice. J Cereb Blood Flow Metab 30:70–78.  https://doi.org/10.1038/jcbfm.2009.185 CrossRefPubMedGoogle Scholar
  24. Hagimori M, Kamiya S, Yamaguchi Y, Arakawa M (2009) Improving frequency of thrombosis by altering blood flow in the carrageenan-induced rat tail thrombosis model. Pharmacol Res 60:320–323.  https://doi.org/10.1016/j.phrs.2009.04.010 CrossRefPubMedGoogle Scholar
  25. Hua Y, Schallert T, Keep RF, Wu J, Hoff JT, Xi G (2002) Behavioral tests after intracerebral hemorrhage in the rat. Stroke 33:2478–2484.  https://doi.org/10.1161/01.STR.0000032302.91894.0F CrossRefPubMedGoogle Scholar
  26. Kakkar P, Das B, Viswanathan PN (1984) A modified spectrophotometric assay of superoxide dismutase. Indian J Biochem Biophys 21:130–132PubMedGoogle Scholar
  27. Khodaverdi E, Khalili N, Zangiabadi F, Homayouni A (2012) Preparation, characterization and stability studies of glassy solid dispersions of indomethacin using PVP and isomalt as carriers. Iran J Basic Med Sci 15:820–832.  https://doi.org/10.22038/IJBMS.2012.4866 PubMedPubMedCentralGoogle Scholar
  28. Kim HW, Kim J, Kim J, Lee S, Choi BR, Han JS, Lee KW, Lee HJ (2013) 3,3′-Diindolylmethane inhibits lipopolysaccharide-induced microglial hyperactivation and attenuates brain inflammation. Toxicol Sci 137(1):158–167.  https://doi.org/10.1093/toxsci/kft240 CrossRefPubMedGoogle Scholar
  29. Krishnamurthy S, Garabadu D, Easwaran S, Bhaseen S, Paliwal P, Rajat V (2015) Silibin in ameliorates the hepatic blood flow in T2dm animals. Indian J Pharmacol 47(1):S57–S177Google Scholar
  30. Kumar G, Paliwal P, Mukherjee S, Patnaik N, Krishnamurthy S, Patnaik R (2018) Pharmacokinetics and brain penetration study of chlorogenic acid. Xenobiotica:1–33.  https://doi.org/10.1080/00498254.2018.1445882
  31. Li J, He J, Du Y, Cui J, Ma Y, Zhang X (2014) Electroacupuncture improves cerebral blood flow and attenuates moderate ischemic injury via angiotensin II its receptors-mediated mechanism in rats. BMC Complement Altern Med 14:441.  https://doi.org/10.1186/1472-6882-14-441 CrossRefPubMedPubMedCentralGoogle Scholar
  32. Lipton P (1999) Ischemic cell death in brain neurons. Physiol Rev 79:1431–1568.  https://doi.org/10.1152/physrev.1999.79.4.1431 CrossRefPubMedGoogle Scholar
  33. Liu F, McCullough LD (2011) Middle cerebral artery occlusion model in rodents: methods and potential pitfalls. Biomed Res Int 2011:1–9.  https://doi.org/10.1155/2011/464701 Google Scholar
  34. Longa EZ, Weinstein PR, Carlson S, Cummins R (1989) Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20:84–91.  https://doi.org/10.1161/01.STR.20.1.84 CrossRefPubMedGoogle Scholar
  35. Luck H (1974) Estimation of catalase activity. Methods of enzymology (Ed Bergmeyer U), Academic Press, New York, 885Google Scholar
  36. Marcus AJ, Broekman MJ, Drosopoulos JH, Islam N, Alyonycheva TN, Safier LB, Hajjar KA, Posnett DN, Schoenborn MA, Schooley KA, Gayle RB (1997) The endothelial cell ecto-ADPase responsible for inhibition of platelet function is CD39. J Clin Invest 99(6):1351–1360.  https://doi.org/10.1172/JCI119294 CrossRefPubMedPubMedCentralGoogle Scholar
  37. Muddana NR, Benade V, Paliwal P, Muppidi AB, Subramanian R, Chillakur M, Kandikere V, Bhyrapuneni G, Nirogi R (2014) Neuro-pharmacokinetics based prediction of p-glycoprotein liability in early drug discovery. Drug Metab Rev 45:111–112Google Scholar
  38. Muddana NR, Paliwal P, Aleti RR, Bhyrapuneni G, Nirogi R (2015) Pharmacokinetics and brain penetration of donepezil after intranasal, oral, intravenous administration using rat as model—a comparison study. Drug Metab Rev 47:227–227Google Scholar
  39. Nazam Ansari M, Bhandari U, Islam F, Tripathi CD (2008) Evaluation of antioxidant and neuroprotective effect of ethanolic extract of Embelia ribes Burm in focal cerebral ischemia/reperfusion induced oxidative stress in rats. Fundam Clin Pharmacol 22:305–314.  https://doi.org/10.1111/j.1472-8206.2008.00580.x CrossRefPubMedGoogle Scholar
  40. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358.  https://doi.org/10.1016/0003-2697(79)90738-3 CrossRefPubMedGoogle Scholar
  41. Paliwal P, Dash D, Krishnamurthy S (2017) Pharmacokinetic study of piracetam in focal cerebral ischemic rats. Eur J Drug Metab Pharmacokinet 15:1–9.  https://doi.org/10.1007/s13318-017-0435-9 Google Scholar
  42. Park MK, Rhee YH, Lee HJ, Lee EO, Kim KH, Park MJ, Jeon BH, Shim BS, Jung CH, Choi SH, Ahn KS (2008) Antiplatelet and antithrombotic activity of indole 3 carbinol in vitro and in vivo. Phytother Res 22:58–64.  https://doi.org/10.1002/ptr.2260 CrossRefPubMedGoogle Scholar
  43. Pinsky DJ, Broekman MJ, Peschon JJ, Stocking KL, Fujita T, Ramasamy R, Connolly ES Jr, Huang J, Kiss S, Zhang Y, Choudhri TF, McTaggart RA, Liao H, Drosopoulos JH, Price VL, Marcus AJ, Maliszewski CR (2002) Elucidation of the thromboregulatory role of CD39/ectoapyrase in the ischemic brain. J Clin Invest 109:1031–1040.  https://doi.org/10.1172/JCI10649 CrossRefPubMedPubMedCentralGoogle Scholar
  44. Sandercock P, Warlow C, Dennis M, Chen ZM (1999) Antiplatelet therapy in acute cerebral ischaemia. Stroke 30(10):2238–2248.  https://doi.org/10.1161/01.STR.30.10.2238k CrossRefGoogle Scholar
  45. Schaar KL, Brenneman MM, Savitz SI (2010) Functional assessments in the rodent stroke model. Exp Transl Stroke Med 2:13.  https://doi.org/10.1186/2040-7378-2-13 CrossRefPubMedPubMedCentralGoogle Scholar
  46. Shakya M, Paliwal P, Patil S, Koti BC, Swamy AH (2011) Cardioprotective effect of'Qolest'a polyherbal formulation against doxorubicin induced cardiotoxicity in wistar rats. Int J Res Pharm Sci 1(3):85–100Google Scholar
  47. Shirley R, Ord EN, Work LM (2014) Oxidative stress and the use of antioxidants in stroke. Antioxidants 3:472–501.  https://doi.org/10.3390/antiox3030472 CrossRefPubMedPubMedCentralGoogle Scholar
  48. Singh S, Sathali AA, Jayaswal SB (2002) Improvement of dissolution rate and aqueous solubility of nitrazepam by solid dispersion technique. Acta Pharm Turc 44:105–118Google Scholar
  49. Thiyagarajan M, Sharma SS (2004) Neuroprotective effect of curcumin in middle cerebral artery occlusion induced focal cerebral ischemia in rats. Life Sci 74:969–985.  https://doi.org/10.1016/j.lfs.2003.06.042 CrossRefPubMedGoogle Scholar
  50. Tripathi A, Paliwal P, Krishnamurthy S (2017) Piracetam attenuates LPS-induced neuroinflammation and cognitive impairment in rats. Cell Mol Neurobiol 37(8):1373–1386.  https://doi.org/10.1007/s10571-017-0468-2 CrossRefPubMedGoogle Scholar
  51. Tsai MJ, Tsai SK, Huang MC, Liou DY, Huang SL, Hsieh WH, Huang WC, Huang SS, Cheng H (2015) Acidic FGF promotes neurite outgrowth of cortical neurons and improves neuroprotective effect in a cerebral ischemic rat model. Neuroscience 305:238–247.  https://doi.org/10.1016/j.neuroscience.2015.07.074 CrossRefPubMedGoogle Scholar
  52. Weaver J, Liu KJ (2015) Does normobaric hyperoxia increase oxidative stress in acute ischemic stroke? A critical review of the literature. Med Gas Res 5:11.  https://doi.org/10.1186/s13618-015-0032-4 CrossRefPubMedPubMedCentralGoogle Scholar
  53. Yamauchi K, Imai T, Shimazawa M, Iwama T, Hara H (2017) Effects of ticagrelor in a mouse model of ischemic stroke. Sci Rep 7(1):12088.  https://doi.org/10.1038/s41598-017-12205-w CrossRefPubMedPubMedCentralGoogle Scholar
  54. Yang L, Chen X, Wang S, Fei Y, Wang D, Li Y, He G, Wu Q, Chu S, Fang W (2015) N2 extenuates experimental ischemic stroke through platelet aggregation inhibition. Thromb Res 136:1310–1317.  https://doi.org/10.1016/j.thromres.2015.10.039 CrossRefPubMedGoogle Scholar
  55. Yousuf S, Salim S, Ahmad M, Ahmed AS, Ansari MA, Islam F (2005) Protective effect of Khamira Abresham Uood Mastagiwala against free radical induced damage in focal cerebral ischemia. J Ethnopharmacol 99:179–184.  https://doi.org/10.1016/j.jep.2004.12.035 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Neurotherapeutics Laboratory, Department of Pharmaceutical Engineering and TechnologyIndian Institute of Technology (Banaras Hindu University)VaranasiIndia
  2. 2.Department of Biochemistry, Institute of Medical SciencesBanaras Hindu UniversityVaranasiIndia
  3. 3.Department of Pathology, Institute of Medical SciencesBanaras Hindu UniversityVaranasiIndia

Personalised recommendations