Effect of different protocols of ischemic postconditioning on skeletal muscle ischemia/reperfusion injury in a rat model

doi:10.3969/j.issn.2095-4344.2015.05.015

Abstract

Abstract:

BACKGROUND: Recent studies have shown that ischemic postconditioning can trigger endogenous protection against ischemia/reperfusion injury in various organs including myocardium and skeletal muscle. However, there is variability in protective effects with different protocols and species specificity.
OBJECTIVE: To investigate effects of various ischemic postconditioning protocols on rat skeletal muscle ischemia/reperfusion injury and to select an optimal protocol.
METHODS: Healthy Adult male Sprague-Dawley rats were randomly allocated into four groups(n = 9): one ischemia/reperfusion injury group and three ischemic postconditioning groups. The rats in ischemia/reperfusion injury group were subjected to 4 hours of main ischemia in the right lower limbs via occlusion of the femoral artery with a clamp, followed by 24 hours of reperfusion, and sham surgery in the left lower limbs as self control. The surgical procedures in three ischemic postconditioning groups were same as in ischemia/reperfusion group except additional ischemic postconditioning interventions were applied at the onset of reperfusion after 4 hours of ischemia with three different algorithms respectively: 4 cycles of 10 seconds reperfusion/10 seconds ischemia, 4 cycles of 30 seconds reperfusion/30 seconds ischemia, and 4 cycles of 1 minute reperfusion/1 minute ischemia. At the end of 24-hr reperfusion in each group, blood samples were taken from femoral vein for detecting serum lactate dehydrogenase(LDH), and tissue samples from gastrocnemius for measuring wet-dry weight (wet/dry)ratio, enzyme myeloperoxidase and malondialdehyde. Tibialis anterior muscle samples were also collected for observing the pathological changes in skeletal muscle under electron microscope.
RESULTS AND CONCLUSION: The wet/dry ratio was significantly lower in the 30-second reperfusion/30-second ischemia group than in the ischemia/reperfusion injury group (P < 0.05). No significant difference was detected between other two ischemic postconditioning groups and ischemia/reperfusion group (P > 0.05). Plasma lactate dehydrogenase activity, gastrocnemius myeloperoxidase activity and malondialdehyde contents were lower in the three ischemic postconditioning groups compared to the ischemia/reperfusion injury group (P < 0.05). No significant difference was detectable among the three ischemic postconditioning groups. Vacuolar degeneration of skeletal muscle mitochondria, myofibrils structural clarity and integrity of the nucleus were apparently improved in the 30-second reperfusion/30-second ischemia group compared to the ischemia/reperfusion injury group under electron microscope. Ultrastructure changes of the other two ischemic postconditioning groups were improved to different degrees as compared with the ischemia/reperfusion injury group. The results indicate that ischemic postconditioning confers protection against skeletal muscle ischemia/reperfusion injury in the rat model. The protocol of four cycle of 30-second reperfusion/30-second ischemia is optimal, which can be used as a basis for further investigations.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: Ischemia Reperfusion, Skeletal Muscle, Lactate Dehydrogenases, Myeloperoxidase, Malondialdehyde

CLC Number:

R318

Peng Long-long, Yang Fu-chun, Bo Zhan-dong, Tan Zhen, Yao Jun, Cheng Jian-wen, Xue Ming-qiang, Zhao Jin-min. Effect of different protocols of ischemic postconditioning on skeletal muscle ischemia/reperfusion injury in a rat model[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(5): 739-744.

Figures/Tables 3

References

[1] Rubin BB, Romaschin A, Walker PM, et al. Mechanisms of postischemic injury in skeletal muscle: intervention strategies . J Appl Physiol (1985).1996;80(2):369-387.
[2] Eckert P, Schnackerz K. Ischemic tolerance of human skeletal muscle. Ann plast surg.1991;26(1):77-84.
[3] Murry CE, Jennings RB, Reimer KA. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation.1986;74(5):1124-1136.
[4] Otani H, Kawasaki H, Ninomiya H, et al. Effects of ischemic preconditioning on the recovery of myocardial function after unprotected ischemia and cardioplegia in the isolated and crystalloid perfused rat hearts. Nihon Kyobu Geka Gakkai. 1997;45(1): 23-30.
[5] Zhang Q, Yan S, Tian Y, et al. Ischemic preconditioning improves liver tolerance to congestion-reperfusion injury in mice. J Surg Res.2014;189(1): 152-158.
[6] Cao C, Wang S, Fan L, et al. Renal protection by ischemic preconditioning is associated with p50/p50 homodimers. Am J Nephrol.2010;31(1): 1-8.
[7] Xia DY, Li W, Qian HR, et al. Ischemia preconditioning is neuroprotective in a rat cerebral ischemic injury model through autophagy activation and apoptosis inhibition. Braz J Med Biol Res. 2013;46(7): 580-8.
[8] Mallick IH, Yang W, Winslet MC, et al. Protective effects of ischemic preconditioning on the intestinal mucosal microcirculation following ischemia-reperfusion of the intestine. Microcirculation.2005;12(8): 615-25.
[9] Saita Y, Yokoyama K, Nakamura K, et al. Protective effect of ischaemic preconditioning against ischaemia-induced reperfusion injury of skeletal muscle: how many preconditioning cycles are appropriate?. Br J Plast Surg. 2002; 55(3): 241-245.
[10] Mansour Z, Bouitbir J, Charles AL, et al. Remote and local ischemic preconditioning equivalently protects rat skeletal muscle mitochondrial function during experimental aortic cross-clamping. J Vascul surg.2012;55(2): 497-505 e1.
[11] 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. 2003;285(2): H579-88.
[12] Gulati P, Singh N. Pharmacological evidence for connection of nitric oxide-mediated pathways in neuroprotective mechanism of ischemic postconditioning in mice. J Pharm Bioallied Sci. 2014;6(4): 233-240.
[13] Young SB, Pires AR, Boaventura GT, et al. Effect of ischemic preconditioning and postconditioning on liver regeneration in prepubertal rats. Transplant Proc. 2014;46(6):1867-1871.
[14] Cao GY, Xie RQ, Cui W, et al. Cardiac protection from myocardial ischemic postconditioning and remote postconditioning during myocardial ischemia/reperfusion injury in rabbits. Zhongguo ying yong sheng li xue za zhi. 2012;28(5): 476-80.
[15] Zhang ZY, Liu XH, Guo XS, et al. Calreticulin is involved in ischemic postconditioning-induced attenuation of ischemia/reperfusion injury in rat skeletal muscle. Sheng li xue bao.2007;59(5): 643-650.
[16] Mcallister SE, Ashrafpour H, Cahoon N, et al. Postconditioning for salvage of ischemic skeletal muscle from reperfusion injury: efficacy and mechanism. Am J Physiol Regul Integr Comp Physiol. 2008;295(2): R681-9.
[17] Yan H, Zhang F, Kochevar AJ, et al. The effect of postconditioning on the muscle flap survival after ischemia- reperfusion injury in rats. J Invest Surg.2010;23(5): 249-256.
[18] Kin H, Zhao ZQ, Sun HY, et al. Postconditioning attenuates myocardial ischemia-reperfusion injury by inhibiting events in the early minutes of reperfusion. Cardiovasc Res.2004;62(1): 74-85.
[19] Iliodromitis EK, Georgiadis M, Cohen MV, et al. Protection from post-conditioning depends on the number of short ischemic insults in anesthetized pigs. Basic research in cardiology. 2006;101(6): 502-507.
[20] Van Vuuren D, Lochner A. Ischaemic postconditioning: from bench to bedside. Cardiovasc J Afr 2008;19(6): 311-320.
[21] Lintz JA, Dalio MB, Joviliano EE, et al. Ischemic pre and postconditioning in skeletal muscle injury produced by ischemia and reperfusion in rats. Acta Cir bras.2013;28(6): 441-446.
[22] Mansour Z, Charles AL, Bouitbir J, et al. Remote and local ischemic postconditioning further impaired skeletal muscle mitochondrial function after ischemia-reperfusion. J Vascul Surg.2012;56(3): 774-82 e1.
[23] Carter WO, Bull C, Bortolon E, et al. A murine skeletal muscle ischemia-reperfusion injury model: differential pathology in BALB/c and DBA/2N mice. J Appl Physiol (1985).1998;85(5): 1676-1683.
[24] Smith JK, Grisham MB, Granger DN, et al. Free radical defense mechanisms and neutrophil infiltration in postischemic skeletal muscle. Am J Physiol.1989;256(3 Pt 2): H789-93.
[25] Park JW, Kang JW, Jeon WJ, et al. Postconditioning protects skeletal muscle from ischemia-reperfusion injury. Microsurgery. 2010;30(3): 223-229.
[26] Zheng W, Huang LZ, Zhao L, et al. Superoxide dismutase activity and malondialdehyde level in plasma and morphological evaluation of acute severe hemorrhagic shock in rats. Am J Emerg Med. 2008;26(1): 54-58.
[27] Hu Y. Experimental study on the role of oxygen-free radicals in ischemic reperfusion tissue injury is island skin flaps . Zhonghua kou qiang yi xue za zhi.1994;29(2): 79-81, 127-128.
[28] Lee KR, Cronenwett JL, Shlafer M, et al. Effect of superoxide dismutase plus catalase on Ca2+ transport in ischemic and reperfused skeletal muscle.J Surg Res.1987; 42(1): 24-32.
[29] Seyama A. The role of oxygen-derived free radicals and the effect of free radical scavengers on skeletal muscle ischemia/reperfusion injury. Surg today.1993;23(12): 1060-1067.
[30] Charles AL, Guilbert AS, Bouitbir J, et al. Effect of postconditioning on mitochondrial dysfunction in experimental aortic cross-clamping. Bri J Surg.2011; 98(4): 511-516.