Indole-3-carbinol improves neurobehavioral symptoms in a cerebral ischemic stroke model
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.
KeywordsIndole-3-carbinol Cerebral ischemia Middle cerebral artery occlusion Antithrombotic Cerebral blood flow
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 was obtained from all individual participants included in the study.
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- Luck H (1974) Estimation of catalase activity. Methods of enzymology (Ed Bergmeyer U), Academic Press, New York, 885Google Scholar
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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