Inhibition of Rho-kinase by fasudil restores the cardioprotection of ischemic postconditioninng in hypercholesterolemic rat heart

Wu,Nan; Li,Wenna; Shu,Wenqi; Lv,Yan; Jia,Dalin

doi:10.3892/mmr.2014.2566

Inhibition of Rho-kinase by fasudil restores the cardioprotection of ischemic postconditioninng in hypercholesterolemic rat heart

Authors:
- Nan Wu
- Wenna Li
- Wenqi Shu
- Yan Lv
- Dalin Jia
View Affiliations

Affiliations: Department of Cardiology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: September 15, 2014 https://doi.org/10.3892/mmr.2014.2566
Pages: 2517-2524

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Ischemic postconditioning (IPoC) reduces lethal reperfusion injury under normal conditions, but its effectiveness is blocked by hypercholesterolemia. The present study aimed to determine whether the inhibition of Rho‑kinase by fasudil restores the cardioprotection of IPoC in the hypercholesterolemic rat heart, and to elucidate the potential mechanisms underlying this process. The isolated rat hearts underwent 30 min global ischemia and 120 min reperfusion. IPoC was induced by six cycles of 10 sec ischemia and 10 sec reperfusion at the onset of the reperfusion. Fasudil was administered 15 min prior to ischemia, and wortmannin and L‑NAME were administered following IPoC. The myocardial infarct size, apoptosis, myocardial nitric oxide (NO) content and Rho‑kinase activity, as well as the activation of the phosphatidylinositol 3‑kinase/Akt/endothelial nitric oxide synthase (PI3K/Akt/eNOS) pathway, were examined. The results revealed that IPoC and 1 µM fasudil treatment alone failed to reduce the infarct size and apoptosis rate. However, IPoC combined with 1 µM fasudil treatment or 10 µM fasudil treatment alone restored the cardioprotection as evidenced by the decreasing in infarct size and apoptosis rate, whereas it was blocked by the administration of wortmannin or L‑NAME. Furthermore, IPoC combined with 1 µM fasudil treatment also enhanced the phosphorylation of Akt and eNOS and conferred a significant increase in the content of NO. By contrast, no significant improvements were demonstrated in the phosphorylation of Akt and eNOS, as well as myocardial NO content when treated with 1 µM fasudil and IPoC alone. The inhibition of Rho‑kinase by fasudil was able to restore the cardioprotection of IPoC in the hypercholesterolemic rat heart. The underlying mechanisms involved in this process appear to be mediated by the activation of the PI3K/Akt/eNOS signal pathway and an increase in the myocardial NO content.

View Figures

View References

1	Zhao ZQ, Corvera JS, Halkos ME, et al: Inhibition of myocardial injury by ischemic postconditioning during reperfusion: comparison with ischemic preconditioning. Am J Physiol Heart Circ Physiol. 285:H579–H588. 2003.PubMed/NCBI
2	Schwartz LM and Lagranha CJ: Ischemic postconditioning during reperfusion activates Akt and ERK without protecting against lethal myocardial ischemia-reperfusion injury in pigs. Am J Physiol Heart Circ Physiol. 290:H1011–H1018. 2006. View Article : Google Scholar
3	Gross GJ, Gauthier KM, Moore J, et al: Evidence for role of epoxyeicosatrienoic acids in mediating ischemic preconditioning and postconditioning in dog. Am J Physiol Heart Circ Physiol. 297:H47–H52. 2009. View Article : Google Scholar : PubMed/NCBI
4	Staat P, Rioufol G, Piot C, et al: Postconditioning the human heart. Circulation. 112:2143–2148. 2005. View Article : Google Scholar : PubMed/NCBI
5	Iliodromitis EK, Zoga A, Vrettou A, et al: The effectiveness of postconditioning and preconditioning on infarct size in hypercholesterolemic and normal anesthetized rabbits. Atherosclerosis. 188:356–362. 2006. View Article : Google Scholar : PubMed/NCBI
6	Zhao JL, Yang YJ, You SJ, Cui CJ and Gao RL: Different effects of postconditioning on myocardial no-reflow in the normal and hypercholesterolemic mini-swines. Microvasc Res. 73:137–142. 2007. View Article : Google Scholar : PubMed/NCBI
7	Kupai K, Csonka C, Fekete V, et al: Cholesterol diet-induced hyperlipidemia impairs the cardioprotective effect of postconditioning: role of peroxynitrite. Am J Physiol Heart Circ Physiol. 297:H1729–H1735. 2009. View Article : Google Scholar : PubMed/NCBI
8	Lauzier B, Delemasure S, Pesant M, et al: A cholesterol-rich diet improves resistance to ischemic insult in mouse hearts but suppresses the beneficial effect of post-conditioning. J Heart Lung Transplant. 28:821–826. 2009. View Article : Google Scholar
9	Andreadou I, Farmakis D, Prokovas E, et al: Short-term statin administration in hypercholesterolaemic rabbits resistant to postconditioning: effects on infarct size, endothelial nitric oxide synthase, and nitro-oxidative stress. Cardiovasc Res. 94:501–509. 2012. View Article : Google Scholar
10	Zhang FJ, Ma LL, Wang WN, et al: Hypercholesterolemia abrogates sevoflurane-induced delayed preconditioning against myocardial infarct in rats by alteration of nitric oxide synthase signaling. Shock. 37:485–491. 2012. View Article : Google Scholar
11	Ma LL, Zhang FJ, Qian LB, et al: Hypercholesterolemia blocked sevoflurane-induced cardioprotection against ischemia-reperfusion injury by alteration of the MG53/RISK/GSK3β signaling. Int J Cardiol. 168:3671–3678. 2013.PubMed/NCBI
12	Fukumoto Y, Mohri M, Inokuchi K, et al: Anti-ischemic effects of fasudil, a specific Rho-kinase inhibitor, in patients with stable effort angina. J Cardiovasc Pharmacol. 49:117–121. 2007. View Article : Google Scholar : PubMed/NCBI
13	Wu DJ, Xu JZ, Wu YJ, et al: Effects of fasudil on early atherosclerotic plaque formation and established lesion progression in apolipoprotein E-knockout mice. Atherosclerosis. 207:68–73. 2009. View Article : Google Scholar : PubMed/NCBI
14	Ho TJ, Huang CC, Huang CY and Lin WT: Fasudil, a Rho-kinase inhibitor, protects against excessive endurance exercise training-induced cardiac hypertrophy, apoptosis and fibrosis in rats. Eur J Appl Physiol. 112:2943–2955. 2012. View Article : Google Scholar
15	Wang N, Guan P, Zhang JP, et al: Fasudil hydrochloride hydrate, a Rho-kinase inhibitor, suppresses isoproterenol-induced heart failure in rats via JNK and ERK1/2 pathways. J Cell Biochem. 112:1920–1929. 2011. View Article : Google Scholar : PubMed/NCBI
16	Wolfrum S, Dendorfer A, Rikitake Y, et al: Inhibition of Rho-kinase leads to rapid activation of phosphatidylinositol 3-kinase/protein kinase Akt and cardiovascular protection. Arterioscler Thromb Vasc Biol. 24:1842–1847. 2004. View Article : Google Scholar : PubMed/NCBI
17	Li Y, Zhu W, Tao J, et al: Fasudil protects the heart against ischemia-reperfusion injury by attenuating endoplasmic reticulum stress and modulating SERCA activity: the differential role for PI3K/Akt and JAK2/STAT3 signaling pathways. PLoS One. 7:e481152012. View Article : Google Scholar
18	Jiang ZH, Zhang TT and Zhang JF: Protective effects of fasudil hydrochloride post-conditioning on acute myocardial ischemia/reperfusion injury in rats. Cardiol J. 20:197–202. 2013. View Article : Google Scholar : PubMed/NCBI
19	Zhao H, Wang Y, Wu Y, et al: Hyperlipidemia does not prevent the cardioprotection by postconditioning against myocardial ischemia/reperfusion injury and the involvement of hypoxia inducible factor-1alpha upregulation. Acta Biochim Biophys Sin (Shanghai). 41:745–753. 2009. View Article : Google Scholar
20	Lv Y, Ren Y, Sun L, Wang S, Wei M and Jia D: Protective effect of Na⁽⁺⁾/Ca⁽²⁺⁾ exchange blocker KB-R7943 on myocardial ischemia-reperfusion injury in hypercholesterolemic rats. Cell Biochem Biophys. 66:357–363. 2013.
21	Byrne CJ, McCafferty K, Kieswich J, et al: Ischemic conditioning protects the uremic heart in a rodent model of myocardial infarction. Circulation. 125:1256–1265. 2012. View Article : Google Scholar
22	Ferdinandy P, Schulz R and Baxter GF: Interaction of cardiovascular risk factors with myocardial ischemia/reperfusion injury, preconditioning, and postconditioning. Pharmacol Rev. 59:418–458. 2007. View Article : Google Scholar : PubMed/NCBI
23	Donato M, D’Annunzio V, Berg G, et al: Ischemic postconditioning reduces infarct size by activation of A1 receptors and KATP channels in both normal and hypercholesterolemic rabbits. J Cardiovasc Pharmacol. 49:287–292. 2007. View Article : Google Scholar : PubMed/NCBI
24	Iliodromitis EK, Andreadou I, Prokovas E, et al: Simvastatin in contrast to postconditioning reduces infarct size in hyperlipidemic rabbits: possible role of oxidative/nitrosative stress attenuation. Basic Res Cardiol. 105:193–203. 2010. View Article : Google Scholar
25	Noma K, Oyama N and Liao JK: Physiological role of ROCKs in the cardiovascular system. Am J Physiol Cell Physiol. 290:C661–C668. 2006. View Article : Google Scholar : PubMed/NCBI
26	Hamid SA, Bower HS and Baxter GF: Rho kinase activation plays a major role as a mediator of irreversible injury in reperfused myocardium. Am J Physiol Heart Circ Physiol. 292:H2598–H2606. 2007. View Article : Google Scholar : PubMed/NCBI
27	Jugdutt BI: Nitric oxide and cardioprotection during ischemia-reperfusion. Heart Fail Rev. 7:391–405. 2002. View Article : Google Scholar : PubMed/NCBI
28	Suematsu Y, Ohtsuka T, Hirata Y, et al: L-Arginine given after ischaemic preconditioning can enhance cardioprotection in isolated rat hearts. Eur J Cardiothorac Surg. 19:873–879. 2001. View Article : Google Scholar
29	Izhar U, Schwalb H, Borman JB and Merin G: Cardioprotective effect of L-arginine in myocardial ischemia and reperfusion in an isolated working rat heart model. J Cardiovasc Surg (Torino). 39:321–329. 1998.PubMed/NCBI
30	Dimmeler S, Fleming I, Fisslthaler B, et al: Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature. 399:601–605. 1999. View Article : Google Scholar : PubMed/NCBI
31	Dauber IM, Lesnefsky EJ, VanBenthuysen RM, Weil JV and Horwitz ID: Reactive oxygen metabolite scanvengers decrease functional coronary microvascular injury due to ischemia reperfusion. Am J Physiol. 260:H42–H49. 1991.
32	Wilson SH, Simari RD, Best PJ, et al: Simvastatin preserves coronary endothelial function in hypercholesterolemia in the absence of lipid lowering. Arterioscler Thromb Vasc Biol. 21:122–128. 2001. View Article : Google Scholar : PubMed/NCBI
33	Freixa X, Bellera N, Ortiz-Pérez JT, et al: Ischaemic postconditioning revisited: lack of effects on infarct size following primary percutaneous coronary intervention. Eur Heart J. 33:103–112. 2012. View Article : Google Scholar
34	Tarantini G, Favaretto E, Marra MP, et al: Postconditioning during coronary angioplasty in acute myocardial infarction: the POST-AMI trial. Int J Cardiol. 162:33–38. 2012. View Article : Google Scholar : PubMed/NCBI