Both ischemic preconditioning and ghrelin administration protect hippocampus from ischemia/reperfusion and upregulate uncoupling protein-2

BMC Physiology - Tập 9 Số 1 - 2009
Yajun Liu1, Lian-Bi Chen1, Xiaoqun Xu2, Éric Vicaut3, R. Sercombe3
1Institute of Physiology, School of Medicine, Shandong University, Jinan, 250012, Shandong, PR China
2Institute of Basic Medicine, Shandong Academy of Medical Sciences, Jinan, 250012, Shandong, PR China
3Laboratory of Microcirculation Research (EA 3509), University Paris 7, France

Tóm tắt

Abstract Background A major endogenous protective mechanism in many organs against ischemia/reperfusion (I/R) injury is ischemic preconditioning (IPC). By moderately uncoupling the mitochondrial respiratory chain and decreasing production of reactive oxygen species (ROS), IPC reduces apoptosis induced by I/R by reducing cytochrome c release from the mitochondria. One element believed to contribute to reduce ROS production is the uncoupling protein UCP2 (and UCP3 in the heart). Although its implication in IPC in the brain has been shown in vitro, no in vivo study of protein has shown its upregulation. Our first goal was to determine in rat hippocampus whether UCP2 protein upregulation was associated with IPC-induced protection and increased ROS production. The second goal was to determine whether the peptide ghrelin, which possesses anti-oxidant and protective properties, alters UCP2 mRNA levels in the same way as IPC during protection. Results After global forebrain ischemia (15 min) with 72 h reperfusion (I/R group), we found important neuronal lesion in the rat hippocampal CA1 region, which was reduced by a preceding 3-min preconditioning ischemia (IPC+I/R group), whereas the preconditioning stimulus alone (IPC group) had no effect. Compared to control, UCP2 protein labelling increased moderately in the I/R (+39%, NS) and IPC+I/R (+28%, NS) groups, and substantially in the IPC group (+339%, P < 0.05). Treatment with superoxide dismutase (10000 U/kg ip) at the time of a preconditioning ischemia greatly attenuated (-73%, P < 0.001) the increase in UCP2 staining at 72 h, implying a role of oxygen radicals in UCP2 induction. Hippocampal UCP2 mRNA showed a moderate increase in I/R (+33%, P < 0.05) and IPC+I/R (+40%, P < 0.05) groups versus control, and a large increase in the IPC group (+333%, P < 0.001). In ghrelin experiments, the I/R+ghrelin group (3 daily administrations) showed considerable protection of CA1 neurons versus I/R animals, and increased hippocampal UCP2 mRNA (+151%, P < 0.001). Conclusion We confirm that IPC causes increased expression of UCP2 protein in vivo, at a moment appropriate for protection against I/R in the hippocampus. The two dissimilar protective strategies, IPC and ghrelin administration, were both associated with upregulated UCP2, suggesting that UCP2 may often represent a final common pathway in protection from I/R.

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Tài liệu tham khảo

Barone FC, White RF, Spera PA, Ellison J, Currie RW, Wang X, Feuerstein GZ: Ischemic preconditioning and brain tolerance: temporal histological and functional outcomes, protein synthesis requirement, and interleukin-1 receptor antagonist and early gene expression. Stroke. 1998, 29: 1937-1950. discussion 1950-1931

Sack MN: Mitochondrial depolarization and the role of uncoupling proteins in ischemia tolerance. Cardiovasc Res. 2006, 72: 210-219. 10.1016/j.cardiores.2006.07.010.

Dirnagl U, Meisel A: Endogenous neuroprotection: mitochondria as gateways to cerebral preconditioning?. Neuropharmacology. 2008, 55: 334-344. 10.1016/j.neuropharm.2008.02.017.

Sommer C: Neuronal plasticity after ischemic preconditioning and TIA-like preconditioning ischemic periods. Acta Neuropathol. 2009, 117: 511-523. 10.1007/s00401-008-0473-0.

Hausenloy DJ, Yellon DM: Preconditioning and postconditioning: Underlying mechanisms and clinical application. Atherosclerosis. 2009, 204: 334-341. 10.1016/j.atherosclerosis.2008.10.029.

Zhan RZ, Wu C, Fujihara H, Taga K, Qi S, Naito M, Shimoji K: Both caspase-dependent and caspase-independent pathways may be involved in hippocampal CA1 neuronal death because of loss of cytochrome c From mitochondria in a rat forebrain ischemia model. J Cereb Blood Flow Metab. 2001, 21: 529-540. 10.1097/00004647-200105000-00007.

Morin C, Simon N: Mitochondria: a target for neuroprotective interventions in cerebral ischemia-reperfusion. Curr Pharm Des. 2006, 12: 739-757. 10.2174/138161206775474242.

Liu Y, Sercombe R, Xie D, Liu K, Chen L: Inhibition of caspase-9 activation and apoptosis is involved in ischemic preconditioning-induced neuroprotection in rat brain. Neurol Res. 2007, 29: 855-861. 10.1179/016164107X223575.

Wang JY, Shen J, Gao Q, Ye ZG, Yang SY, Liang HW, Bruce IC, Luo BY, Xia Q: Ischemic postconditioning protects against global cerebral ischemia/reperfusion-induced injury in rats. Stroke. 2008, 39: 983-990. 10.1161/STROKEAHA.107.499079.

Lopez-Neblina F, Toledo AH, Toledo-Pereyra LH: Molecular biology of apoptosis in ischemia and reperfusion. J Invest Surg. 2005, 18: 335-350. 10.1080/08941930500328862.

Chen SD, Wu HY, Yang DI, Lee SY, Shaw FZ, Lin TK, Liou CW, Chuang YC: Effects of rosiglitazone on global ischemia-induced hippocampal injury and expression of mitochondrial uncoupling protein 2. Biochem Biophys Res Commun. 2006, 351: 198-203. 10.1016/j.bbrc.2006.10.017.

Chen Q, Moghaddas S, Hoppel CL, Lesnefsky EJ: Ischemic defects in the electron transport chain increase the production of reactive oxygen species from isolated rat heart mitochondria. Am J Physiol Cell Physiol. 2008, 294: C460-466. 10.1152/ajpcell.00211.2007.

Liem DA, Manintveld OC, Schoonderwoerd K, McFalls EO, Heinen A, Verdouw PD, Sluiter W, Duncker DJ: Ischemic preconditioning modulates mitochondrial respiration, irrespective of the employed signal transduction pathway. Transl Res. 2008, 151: 17-26. 10.1016/j.trsl.2007.09.007.

McLeod CJ, Aziz A, Hoyt RF, McCoy JP, Sack MN: Uncoupling proteins 2 and 3 function in concert to augment tolerance to cardiac ischemia. J Biol Chem. 2005, 280: 33470-33476. 10.1074/jbc.M505258200.

Mattiasson G, Shamloo M, Gido G, Mathi K, Tomasevic G, Yi S, Warden CH, Castilho RF, Melcher T, Gonzalez-Zulueta M, et al: Uncoupling protein-2 prevents neuronal death and diminishes brain dysfunction after stroke and brain trauma. Nat Med. 2003, 9: 1062-1068. 10.1038/nm903.

Diano S, Matthews RT, Patrylo P, Yang L, Beal MF, Barnstable CJ, Horvath TL: Uncoupling protein 2 prevents neuronal death including that occurring during seizures: a mechanism for preconditioning. Endocrinology. 2003, 144: 5014-5021. 10.1210/en.2003-0667.

Fujimura M, Morita-Fujimura Y, Kawase M, Copin JC, Calagui B, Epstein CJ, Chan PH: Manganese superoxide dismutase mediates the early release of mitochondrial cytochrome C and subsequent DNA fragmentation after permanent focal cerebral ischemia in mice. J Neurosci. 1999, 19: 3414-3422.

Kim GW, Kondo T, Noshita N, Chan PH: Manganese superoxide dismutase deficiency exacerbates cerebral infarction after focal cerebral ischemia/reperfusion in mice: implications for the production and role of superoxide radicals. Stroke. 2002, 33: 809-815. 10.1161/hs0302.103745.

Paradis E, Clavel S, Bouillaud F, Ricquier D, Richard D: Uncoupling protein 2: a novel player in neuroprotection. Trends Mol Med. 2003, 9: 522-525. 10.1016/j.molmed.2003.10.009.

Olsson T, Wieloch T, Diano S, Warden CH, Horvath TL, Mattiasson G: Overexpression of UCP2 protects thalamic neurons following global ischemia in the mouse. J Cereb Blood Flow Metab. 2008, 28: 1186-1195. 10.1038/jcbfm.2008.8.

Richard D, Clavel S, Huang Q, Sanchis D, Ricquier D: Uncoupling protein 2 in the brain: distribution and function. Biochem Soc Trans. 2001, 29: 812-817. 10.1042/BST0290812.

Brand MD, Affourtit C, Esteves TC, Green K, Lambert AJ, Miwa S, Pakay JL, Parker N: Mitochondrial superoxide: production, biological effects, and activation of uncoupling proteins. Free Radic Biol Med. 2004, 37: 755-767. 10.1016/j.freeradbiomed.2004.05.034.

Echtay KS, Brand MD: 4-hydroxy-2-nonenal and uncoupling proteins: an approach for regulation of mitochondrial ROS production. Redox Rep. 2007, 12: 26-29. 10.1179/135100007X162158.

Duval C, Negre-Salvayre A, Dogilo A, Salvayre R, Penicaud L, Casteilla L: Increased reactive oxygen species production with antisense oligonucleotides directed against uncoupling protein 2 in murine endothelial cells. Biochem Cell Biol. 2002, 80: 757-764. 10.1139/o02-158.

Brand MD, Esteves TC: Physiological functions of the mitochondrial uncoupling proteins UCP2 and UCP3. Cell Metab. 2005, 2: 85-93. 10.1016/j.cmet.2005.06.002.

Kojima M, Kangawa K: Ghrelin: structure and function. Physiol Rev. 2005, 85: 495-522. 10.1152/physrev.00012.2004.

Cao JM, Ong H, Chen C: Effects of ghrelin and synthetic GH secretagogues on the cardiovascular system. Trends Endocrinol Metab. 2006, 17: 13-18. 10.1016/j.tem.2005.11.004.

Benso A, Broglio F, Marafetti L, Lucatello B, Seardo MA, Granata R, Martina V, Papotti M, Muccioli G, Ghigo E: Ghrelin and synthetic growth hormone secretagogues are cardioactive molecules with identities and differences. Semin Vasc Med. 2004, 4: 107-114. 10.1055/s-2004-835367.

Chang L, Ren Y, Liu X, Li WG, Yang J, Geng B, Weintraub NL, Tang C: Protective effects of ghrelin on ischemia/reperfusion injury in the isolated rat heart. J Cardiovasc Pharmacol. 2004, 43: 165-170. 10.1097/00005344-200402000-00001.

Konturek PC, Brzozowski T, Walter B, Burnat G, Hess T, Hahn EG, Konturek SJ: Ghrelin-induced gastroprotection against ischemia-reperfusion injury involves an activation of sensory afferent nerves and hyperemia mediated by nitric oxide. Eur J Pharmacol. 2006, 536: 171-181. 10.1016/j.ejphar.2006.02.032.

El Eter E, Al Tuwaijiri A, Hagar H, Arafa M: In vivo and in vitro antioxidant activity of ghrelin: Attenuation of gastric ischemic injury in the rat. J Gastroenterol Hepatol. 2007, 22: 1791-1799. 10.1111/j.1440-1746.2006.04696.x.

Wu R, Dong W, Ji Y, Zhou M, Marini CP, Ravikumar TS, Wang P: Orexigenic hormone ghrelin attenuates local and remote organ injury after intestinal ischemia-reperfusion. PLoS ONE. 2008, 3: e2026-10.1371/journal.pone.0002026.

Granata R, Settanni F, Trovato L, Destefanis S, Gallo D, Martinetti M, Ghigo E, Muccioli G: Unacylated as well as acylated ghrelin promotes cell survival and inhibit apoptosis in HIT-T15 pancreatic beta-cells. J Endocrinol Invest. 2006, 29: RC19-22.

Liu Y, Wang PS, Xie D, Liu K, Chen L: Ghrelin reduces injury of hippocampal neurons in a rat model of cerebral ischemia/reperfusion. Chin J Physiol. 2006, 49 (5): 244-250.

Chung H, Kim E, Lee DH, Seo S, Ju S, Lee D, Kim H, Park S: Ghrelin inhibits apoptosis in hypothalamic neuronal cells during oxygen-glucose deprivation. Endocrinology. 2007, 148: 148-159. 10.1210/en.2006-0991.

Nakatsuka H, Ohta S, Tanaka J, Toku K, Kumon Y, Maeda N, Sakanaka M, Sakaki S: Cytochrome c release from mitochondria to the cytosol was suppressed in the ischemia-tolerance-induced hippocampal CA1 region after 5-min forebrain ischemia in gerbils. Neurosci Lett. 2000, 278: 53-56. 10.1016/S0304-3940(99)00894-0.

Miao Y, Xia Q, Hou Z, Zheng Y, Pan H, Zhu S: Ghrelin protects cortical neuron against focal ischemia/reperfusion in rats. Biochem Biophys Res Commun. 2007, 359: 795-800. 10.1016/j.bbrc.2007.05.192.

Tsubone T, Masaki T, Katsuragi I, Tanaka K, Kakuma T, Yoshimatsu H: Ghrelin regulates adiposity in white adipose tissue and UCP1 mRNA expression in brown adipose tissue in mice. Regul Pept. 2005, 130: 97-103. 10.1016/j.regpep.2005.04.004.

Barazzoni R, Bosutti A, Stebel M, Cattin MR, Roder E, Visintin L, Cattin L, Biolo G, Zanetti M, Guarnieri G: Ghrelin regulates mitochondrial-lipid metabolism gene expression and tissue fat distribution in liver and skeletal muscle. Am J Physiol Endocrinol Metab. 2005, 288: E228-235. 10.1152/ajpendo.00115.2004.

Pecqueur C, Alves-Guerra MC, Gelly C, Levi-Meyrueis C, Couplan E, Collins S, Ricquier D, Bouillaud F, Miroux B: Uncoupling protein 2, in vivo distribution, induction upon oxidative stress, and evidence for translational regulation. J Biol Chem. 2001, 276: 8705-8712. 10.1074/jbc.M006938200.

Horvath B, Spies C, Horvath G, Kox WJ, Miyamoto S, Barry S, Warden CH, Bechmann I, Diano S, Heemskerk J, et al: Uncoupling protein 2 (UCP2) lowers alcohol sensitivity and pain threshold. Biochem Pharmacol. 2002, 64: 369-374. 10.1016/S0006-2952(02)01167-X.

Kitagawa K, Matsumoto M, Kuwabara K, Tagaya M, Ohtsuki T, Hata R, Ueda H, Handa N, Kimura K, Kamada T: 'Ischemic tolerance' phenomenon detected in various brain regions. Brain Res. 1991, 561: 203-211. 10.1016/0006-8993(91)91596-S.

Puisieux F, Deplanque D, Bulckaen H, Maboudou P, Gele P, Lhermitte M, Lebuffe G, Bordet R: Brain ischemic preconditioning is abolished by antioxidant drugs but does not up-regulate superoxide dismutase and glutathion peroxidase. Brain Res. 2004, 1027: 30-37. 10.1016/j.brainres.2004.08.067.

Schmidt-Kastner R, Hossmann KA: Distribution of ischemic neuronal damage in the dorsal hippocampus of rat. Acta Neuropathol. 1988, 76: 411-421. 10.1007/BF00686979.

Kitahara T, Li HS, Balaban CD: Localization of the mitochondrial uncoupling protein family in the rat inner ear. Hear Res. 2004, 196: 39-48. 10.1016/j.heares.2004.02.002.

Parre Van De TJ, Martinet W, Verheye S, Kockx MM, Van Langenhove G, Herman AG, De Meyer GR: Mitochondrial uncoupling protein 2 mediates temperature heterogeneity in atherosclerotic plaques. Cardiovasc Res. 2008, 77: 425-431. 10.1093/cvr/cvm003.

Krauss S, Zhang CY, Lowell BB: A significant portion of mitochondrial proton leak in intact thymocytes depends on expression of UCP2. Proc Natl Acad Sci USA. 2002, 99: 118-122. 10.1073/pnas.012410699.

Nakagomi T, Kirino T, Kanemitsu H, Tsujita Y, Tamura A: Early recovery of protein synthesis following ischemia in hippocampal neurons with induced tolerance in the gerbil. Acta Neuropathol. 1993, 86: 10-15. 10.1007/BF00454892.

Busquets S, Alvarez B, Van Royen M, Figueras MT, Lopez-Soriano FJ, Argiles JM: Increased uncoupling protein-2 gene expression in brain of lipopolysaccharide-injected mice: role of tumour necrosis factor-alpha?. Biochim Biophys Acta. 2001, 1499: 249-256. 10.1016/S0167-4889(00)00126-9.

Kim-Han JS, Dugan LL: Mitochondrial uncoupling proteins in the central nervous system. Antioxid Redox Signal. 2005, 7: 1173-1181. 10.1089/ars.2005.7.1173.

Mori T, Muramatsu H, Matsui T, McKee A, Asano T: Possible role of the superoxide anion in the development of neuronal tolerance following ischaemic preconditioning in rats. Neuropathol Appl Neurobiol. 2000, 26: 31-40. 10.1046/j.1365-2990.2000.00215.x.

Preston E, Sutherland G, Finsten A: Three openings of the blood-brain barrier produced by forebrain ischemia in the rat. Neurosci Lett. 1993, 149: 75-78. 10.1016/0304-3940(93)90351-K.

Roda JM, Carceller F, Pajares R, Diez-Tejedor E: Prevention of cerebral ischaemic reperfusion injury by intra-arterial administration of superoxide dismutase in the rat. Neurol Res. 1991, 13: 160-163.

Li L, Zhang LK, Pang YZ, Pan CS, Qi YF, Chen L, Wang X, Tang CS, Zhang J: Cardioprotective effects of ghrelin and des-octanoyl ghrelin on myocardial injury induced by isoproterenol in rats. Acta Pharmacol Sin. 2006, 27: 527-535. 10.1111/j.1745-7254.2006.00319.x.

De Vriese C, Hacquebard M, Gregoire F, Carpentier Y, Delporte C: Ghrelin interacts with human plasma lipoproteins. Endocrinology. 2007, 148: 2355-2362. 10.1210/en.2006-1281.

Wu C, Fujihara H, Yao J, Qi S, Li H, Shimoji K, Baba H: Different expression patterns of Bcl-2, Bcl-xl, and Bax proteins after sublethal forebrain ischemia in C57Black/Crj6 mouse striatum. Stroke. 2003, 34: 1803-1808. 10.1161/01.STR.0000077255.15597.69.

Obay BD, Tasdemir E, Tumer C, Bilgin HM, Atmaca M: Dose dependent effects of ghrelin on pentylenetetrazole-induced oxidative stress in a rat seizure model. Peptides. 2008, 29: 448-455. 10.1016/j.peptides.2007.11.020.

Zhao H, Liu G, Wang Q, Ding L, Cai H, Jiang H, Xin Z: Effect of ghrelin on human endothelial cells apoptosis induced by high glucose. Biochem Biophys Res Commun. 2007, 362: 677-681. 10.1016/j.bbrc.2007.08.021.

Andrews ZB, Liu ZW, Walllingford N, Erion DM, Borok E, Friedman JM, Tschop MH, Shanabrough M, Cline G, Shulman GI, et al: UCP2 mediates ghrelin's action on NPY/AgRP neurons by lowering free radicals. Nature. 2008, 454: 846-851. 10.1038/nature07181.

Hioki C, Yoshida T, Kogure A, Takakura Y, Umekawa T, Yoshioka K, Shimatsu A, Yoshikawa T: Effects of growth hormone (GH) on mRNA levels of uncoupling proteins 1, 2, and 3 in brown and white adipose tissues and skeletal muscle in obese mice. Horm Metab Res. 2004, 36: 607-613. 10.1055/s-2004-825905.

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Tập 9 Số 1

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