Neste estudo utilizou-se dois moduladores que têm seus mecanismos de ação na proteção da lesão de IR ainda não totalmente esclarecidos e que nesta fase da lesão também têm sido menos estudados.
Os resultados evidenciam uma proteção do fígado pela N-acetilcisteína e pelo
precondicionamento isquêmico e também uma proteção ao pulmão de ambos os moduladores na fase tardia da lesão de isquemia e reperfusão hepática.
Esses achados nos permitem sugerir que tanto o IPC quanto a NAC atuam sobre a cascata das caspases inibindo a apoptose. Com ambos os moduladores, também se pode observar que o fígado e o pulmão conseguem manter a indução da proliferação celular, permitindo o processo de manutenção do reparo celular após a lesão de isquemia e reperfusão hepática neste modelo de IR.
A maior necessidade de órgãos para transplante, pelo aumento de doenças crônicas, e o maior número de traumas hepáticos advindos da violência urbana, fazem com estratégias de redução da lesão de isquemia hepática sejam constantemente buscadas.
Estimulados pelos experimentos em animais, alguns grupos de pesquisadores iniciaram recentemente a aplicação do IPC em estudos clínicos. Esses trabalhos têm demonstrado que outros fatores ainda devem ser avaliados. Clavien et al47 em um estudo prospectivo com 100 pacientes onde o IPC foi efetivo em reduzir as lesões de IR evidenciou algumas situações onde o IPC foi menos efetivo. Mais recentemente Azoulay et al49 relataram pela primeira vez o uso do IPC no transplante clinico demonstrando uma maior tolerância à lesão de IR, mas identificaram uma piora da função hepática inicial. A NAC também tem sido usada nos últimos anos em alguns estudos clínicos de IR demonstrando que existe uma necessidade de ajuste no tempo, modo e dose da aplicação. Apesar desses estudos nos últimos anos com o IPC e a NAC, ainda restam muitas dúvidas a serem esclarecidas, tanto no que se refere aos mecanismos de ação quanto aos critérios de utilização.
47
Este estudo desenvolvido em uma linha de pesquisa, que se propõe a estudar o papel do IPC e da NAC na lesão de IR, contribui com a identificação da apoptose como importante mecanismo de morte celular na fase tardia da lesão e com a identificação da proliferação celular neste modelo de IR, além de verificar a proteção do IPC e da NAC no fígado e no pulmão na fase tardia.
48
6. CONCLUSÕES
1. O precondicionamento isquêmico e a N-acetilcisteína protegem o fígado e o
pulmão na fase tardia da lesão de isquemia e reperfusão hepática, segundo a ocorrência de apoptose e proliferação celular.
2. As alterações morfológicas do fígado foram minimizadas por ambas as estratégias estudadas, exceto o infiltrado inflamatório que foi prevenido principalmente pela N-acetilcisteína.
3. As alterações morfológicas do pulmão tenderam a diminuir com ambas as estratégias, entretanto, somente o espessamento do septo alveolar foi prevenido pelo precondicionamento isquêmico.
49
7. REFERÊNCIAS
1. Peralta C, Bulbena O, Xaus C, Prats N, Cutrin J C, Poli G, Gelpi E, Roselló- Catafau J. Ischemic preconditioning: a defense mechanism against the reactive oxygen species generated after hepatic ischemia reperfusion. Transpl 2002; 73(8):1203-1211.
2. Peralta C, Perales JC, Bartrons R, Mitchell C, Gilgenkrantz H, Xaus C, Prats N, Fernandez L, Gelpi E, Panes J, Rosello-Catafau J. The combination of ischemic preconditioning and liver Bcl-2 over expression is a suitable strategy to prevent liver and lung damage after hepatic ischemia-reperfusion. Am J Pathol 2002; 160:2111:2122.
3. Fernandez L, Heredia N, Peralta C, Xaus C, Rosello-Catafau J, Rimola A, Marco A, Serafin A, Deulofeu R, Gelpi E, Grande L. Role of ischemic preconditioning and the portosystemic shunt in the prevention of liver and lung damage after rat liver transplantation. Transplantation 2003; 76:282:289.
4. Daemen M, De Vries B, Buurman WA. Apoptosis and inflammation in renal reperfusion injury. Transplantation 2002; 73:1693-700.
5. Fondevila C, Busuttil RW, Kupiec-Weglinski JW. Hepatic ischemia/reperfusion injury – a fresh look. Exp Mol Pathol 2003; 74:86-93.
6. Glantzounis GK, Salacinski HJ, Yang W, Davidson BR, Seifalian AM. The contemporary role of antioxidant therapy in attenuating liver ischemia- reperfusion injury: A Review. Liver Transpl 2005; 11(9):1031-47.
7. Kumar, Abbas, Fausto. Robbins and Cotran Pathologic Basis of Disease. 7th ed. Elsevier Inc; 2004.
8. Miranda LEC, Viaro F, Ceneviva R, Évora PRB. The experimental basis of hepatic ischemia-reperfusion injury: review. Acta Cir Bras 2004; 19(1):1-12. 9. Fusai G, Glantzounis GK, Hafez T, Yang W, Quaglia A, Sheth H, Kanoria S,
Parkes H, Seifalian A, Davidson BR. N-Acetylcysteine ameliorates the late phase of liver ischaemia/reperfusion injury in the rabbit with hepatic steatosis. Clin Sci 2005; 109(5):465-73.
50
Protective effects of ischemic preconditioning and application of lipoic acid prior to 90 min of hepatic ischemia in a rat model. World J Gastroenterol 2007; 13(27):3692-8.
11. Jaeschke, H. and Farhood, A. Neutrophil and Kupffer cell-induced oxidant stress and ischemia-reperfusion injury in rat liver. Am J Physiol 1991; 260:G355–G362.
12. Lentsch, A. B., Kato, A., Yoshidome, H., McMasters, K. M. and Edwards, M. J. Inflammatory mechanisms and therapeutic strategies for warm hepatic ischemia/reperfusion injury. Hepatology 2000; 32:169–173.
13. Pesonen EJ, Linder N, Raivio KO, et al. Circulating xanthine oxidase and neutrophil activation during human liver transplantation. Gastroenterology 1998; 114: 1009
14. Lopez-Neblina F, Toledo AH, Toledo-Pereyra LH. Molecular biology of apoptosis in ischemia and reperfusion. J Invest Surg 2005; 18:335-50.
15. Toledo-Pereyra LH and Suzuki S. Neutrophils, cytokines and adhesion molecules in hepatic ischemia and reperfusion injury. J Am Coll Surg 1994; 179:758-62.
16. Andrade DR Jr, Souza RB, Santos SA, Andrade DR. Oxygen free radicals and pulmonary disease. JBP 2005; 31:60-1.
17. Anaya-Prado R, Toledo-Pereyra LH, Lentsch AB, Ward PA. Ischemia/ reperfusion injury. J Surg Res 2002; 105:248-58.
18. Fan C, Zwacka RM, Engelhard JF: Therapeutic approaches for ischemia/ reperfusion injury in the liver. J Mol Med 1999; 77:577-592.
19. Riedemann RR, Ward PA. Complement in ischemia reperfusion injury. Am J Pathol 2003; 162:363-7.
20. Vaux D, Silke J. Mammalian mitochondrial IAP-binding proteins. Biochem Biophys Res Commun 2003; 304(3):499-504.
21. Strasser A, O’Connor L, Dixit VM. Apoptosis signaling. Annu Rev Biochem 2000; 69:217-45.
22. Fausto N, Laird A, Webber EM. Liver regeneration 2. Role of growth factors and cytokines in hepatic regeneration. FASEB J 1995; 9:1527-36.
51
liver regeneration. Br J Surg 2002; 89(9):1089-95.
24. Yamada Y, Webber EM, Kirillova I, Peschon JJ, Fausto N. Analysis of liver regeneration in mice lacking type 1 and type 2 tumor necrosis factor receptor Requirement for type 1 but not type 2 receptor. Hepatology 1998; 28(4):959-70. 25. Fausto N. Liver regeneration and repair. Hepatology 2004; 39:1477-87.
26. Debonera F, Aldeguer X, Shen X, Gelman AE, Gao F, Que X, et al. Activation of interleukin-6/STAT3 and liver regeneration following transplantation. J Surg Res 2001; 96:289-95.
27. Rudiger HA, Clavien PA. Tumor necrosis factor alpha, but not Fas, mediates hepatocellular apoptosis in the murine ischemic liver. Gastroenterology 2002; 122(1):202-10.
28. Michalopoulos GK, Defrance MC. Liver regeneration. Science 1997; 276:60- 66.
29. Ishii S, Abe T, Saito T, Tsuchiya T, Kanno H, Miyazawa M, et al. Effects of preconditioning on ischemia/reperfusion injury of hepatocytes determined by immediate early gene transcription. J Hepatobiliary Pancreat Surg 2001; 8(5):461-8.
30. Schlossberg H, Zhang Y, Dudus L, Engslhardt JF. Expression of c-fos and c- jun during hepatocellular remodeling following ischemia/reperfusion in mouse liver. Hepatology 1996; 23:1546-55.
31. Xiao JS, Cai FG, Niu Y, Zhang Y, Xu XL , Ye QF. Preconditioning effects on expression of proto-oncogenes c-fos and c-jun after hepatic ischemia/reperfusion in rats. Hepatobiliary Pancreat Dis Int 2005; 4:197-202 32. Saito T, Matsumoto I, Goto S, Kamada N, Motoki R, Wilce PA. The differential
induction of two immediate early genes, c-fos and c-jun, after systemic hypovolemic shock/resuscitation in the rat liver and kidney. Surg Today 1998; 28:608-61.
33. Bedirli A, Kerem M, Pasaoglu H, Erdem O, Ofluoglu E, Sakrak O. Effects of ischemic preconditionong on regenerative capacity of hepatocyte in the ischemically damage rat livers. J Surg Res 2005; 125:42-8.
34. Weinbroum AA, Rudick V, Ben-Abraham R, Karchevski E. N-acetyl-L-cysteine for preventing lung reperfusion injury after liver ischemia-reperfusion: A
52
possible dual protective mechanism in a dose-response study. Transplantation 2000; 69:853:859.
35. Murray CE, Jenning RB, Reimer KA: Preconditioning with ischemia: a delay in lethal cell injury in ischemic myocardium. Circulation 1986; 74:1124-36.
36. Glanemann M, Vollmar B, Nussler AK, Schaefer T, Neuhaus P, Menger MD. Ischemic preconditioning protects from hepatic ischemia/reperfusion-injury by preservation of microcirculation and mitochondrial redox-state. J Hepatol 2003; 38: 59-66.
37. Lee WY, Lee SM. Ischemic preconditioning protects post-ischemic oxidative damage to mitochondria in rat liver. Shock 2005; 24:370-75.
38. Uhlmann, D., Uhlmann, S. and Spiegel, H. U. Endothelin/nitric oxide balance influences hepatic ischemia-reperfusion injury. J Cardiovasc Pharmacol 2000; 36: S212–S214.
39. Peralta C, Hotter G, Closa D, Prats N, Xaus C, Gelpi E, Rosello-Catafau J. The protective role of adenosine in inducing nitric oxide synthesis in rat liver ischemia preconditioning is mediated by activation of adenosine A2 receptors. Hepatology 1999; 29(1):126-32.
40. Teoh NC, Farrell GC. Hepatic ischemia reperfusion injury: pathogenic mechanisms and basis for hepatoprotection. J Gastroenterol Hepatol. 2003; 18(8):891-902.
41. Yadav SS, Sindram D, Perry DK, Clavien PA. Ischemic preconditioning protects the mouse liver by inhibition of apoptosis through a caspase- dependent pathway. Hepatology 1999; 30: 1223-1231
42. Natori S, Higuchi H, Contreras P, Gores GJ. The caspase inhibitor IDN-6556 prevents caspase activation and apoptosis in sinusoidal endothelial cells during liver preservation injury. Liver Transpl. 2003; 9(3):278-84.
43. Yin XM, Ding WX. Death receptor activation - induced hepatocyte apoptosis and liver injury. Curr Mol Med. 2003; 3(6):491-508.
44. Rudiger HA, Graf R, Clavien PA. Sub-lethal oxidative stress triggers the protective effects of ischemic preconditioning in the mouse liver. J Hepatol 2003; 39: 972-977.
53
Roselló-Catafau J. Preconditioning protects against systemic disorders associated with hepatic ischemia-reperfusion through blockade of tumor necrosis factor-induced P-selectin up-regulation in the rat. Hepatology. 2001; 33(1):100-13.
46. Petrowsky H, McCormack L, Trujillo M, Selzner M, Jochum W, Clavien PA. A prospective, randomized, controlled trial comparing intermittent portal triad clamping versus ischemic preconditioning with continuous clamping for major liver resection. Ann Surg 2006; 244(6):921-8.
47. Clavien PA, Selzner M, Rudiger HA, Graf R, Kadry Z, Rousson V, Jochum W. A prospective randomized study in 100 consecutive patients undergoing major liver resection with versus without ischemic preconditioning. Ann Surg. 2003; 238(6):843-50.
48. Clavien PA, Yadav S, Sindram D. Protective effects of ischemic preconditioning for liver resection performed under inflow occlusion in humans. Ann Surg 2000; 232:155-62.
49. Azoulay D, Lucidi V, Andreani P, Maggi U, Sebagh M, Ichai P, Lemoine A, Adam R, Castaing D. Ischemic preconditioning for major liver resection under vascular exclusion of the liver preserving the caval flow: a randomized prospective study. J Am Coll Surg. 2006; 202(2):203-11.
50. Quireze C Jr, Montero EFS, Leitão RM, Juliano Y, Fagundes DJ, Poli de Figueiredo LF. Ischemic preconditioning prevents apoptotic cell death and necrosis in early and intermediate phases of liver ischemia-reperfusion injury in rats. J Invest Surg 2006; 19:229-236.
51. Montero EF, Quireze C Jr., d’Oliveira DM. Bile duct exclusion from selective vascular inflow occlusion in rat liver: role of ischemic preconditioning and N- acetylcysteine on hepatic reperfusion injury. Transplant Proc 2005; 37:425:427. 52. Weinbroum AA, Kluger Y, Ben Abraham R, Shapira I, Karchevski E, Rudick V.
Lung preconditioning with N-acetyl-L-cysteine prevents reperfusion injury after liver no flow-reflow: a dose-response study. Transplantation 2001; 71(2):300-6. 53. Portella AO, Montero EF, Poli de Figueiredo LF, Bueno AS, Thurow AA, Rodrigues FG. Effects of N-acetylcysteine in hepatic ischemia-reperfusion injury during hemorrhagic shock. Transplant Proc 2004; 36:846:848
54
54. Lauz SHM, Montero EFM, Gomes LF Taha MO, Junqueira VBC, Simões MJ. Avaliação da lesão isquêmica normotérmica do fígado: papel da oclusão do ducto biliar principal e da N-acetilcisteína. Rev. Col. Bras. Cir. 2005; 32:168- 172.
55. Khan AW, Fuller BJ, Shah SR, Davidson BR, Rolles K. A prospective randomized trial of N-acetylcysteine administration during cold preservation of the donor liver for transplantation. Ann Hepatol 2005; 4:121-126.
56. Glantzounis GK, Rocks SA, Sheth H, Knight I, Salacinski HJ, Davidson BR, Winyard PG, Seifalian AM. Formation and role of plasma S-nitrosothiols in liver ischemia-reperfusion injury. Free Radic Biol Med. 2007; 42(6):882-92.
57. Smyrniotis V, Arkadopoulos N, Kostopanagiotou G, Theodoropoulos T, Theodoraki K, Farantos C, Kairi E, Paphiti A. Attenuation of ischemic injury by N-acetylcysteine preconditioning of the liver. J Surg Res. 2005;129(1):31-7. 58. Salim, CS. Avaliação sistêmica da isquemia e reperfusão hepática em ratos
modulada pela N-acetilcísteina. [Tese de Mestrado]. São Paulo (SP): Universidade Federal de São Paulo – Escola Paulista de Medicina; 2001.
59. Chavez-Cartaya R, Jamieson NV, Ramirez P, Marin J, Pino-Chavez G. Free radical scavengers to prevent reperfusion injury following experimental warm liver ischaemia. Is there a real physiological benefit? Transpl Int. 1999; 12(3):213-21.
60. Thies JC, Teklote J, Clauer U, Töx U, Klar E, Hofmann WJ, Herfarth C, Otto G. The efficacy of N-acetylcysteine as a hepatoprotective agent in liver transplantation. Transpl Int. 1998; 11:390-2.
61. Taut FJ, Schmidt H, Zapletal CM, Thies JC, Grube C, Motsch J, Klar E, Martin E. N-acetylcysteine induces shedding of selectins from liver and intestine during orthotopic liver transplantation. Clin Exp Immunol. 2001; 124(2):337-41. 62. Steib A, Freys G, Collin F, Launoy A, Mark G, Boudjema K. Does N-
acetylcysteine improve hemodynamics and graft function in liver transplantation? Liver Transpl Surg. 1998; 4(2):152-7.
63. Ramalho FS, Fernandez-Monteiro I, Rosello-Catafau J, Peralta C. Hepatic microcirculatory failure. Acta Cir Bras 2006; 21: 48-53.
55
response of the hepatocyte to ischemia. Liver Int 2007; 27: 6-16.
65. Eum HA, Cha YN, Lee SM. Necrosis and apoptosis: sequence of liver damage following reperfusion after 60 min ischemia in rats. Pathobiology 2007; 74(1):42-9.
66. Ijaz S, Yang W, Winslet MC, Seifalian AM. The role of nitric oxide in the modulation and tissue oxygenation in an experimentl model of hepatic steatosis. Microvasc Res 2005; 70: 129-36.
67. Kohli V, Selzner M, Madden JF, Bentley RC, Clavien PA. Endothelial cell and hepatocyte deaths occur by apoptosis after ischemia-reperfusion injury in the rat liver. Transplantation 1999; 67:1099-1105.
68. Gerwig T, Meissner H, Bilzer M. Atrial natriuretic peptide preconditioning protects against hepatic preservation injury by attenuating necrotic and apoptotic cell death. J Hepatol 2003; 39:341-8.
69. Jaeschke H. Reperfusion injury after warm ischemia or cold storage of the liver: Role of apoptotic cell death. Transplant Proc 2002; 34:2656-8. 70.
70. Jaeschke H, Lemasters JJ. Apoptosis versus oncotic necrosis in hepatic ischemia/reperfusion injury. Gastroenterology 2003; 125(4):1246-57.
71. Rudiger HA, Graf R, Clavien PA. Liver ischemia: apoptosis as a central mechanism of injury. J Invest Surg. 2003; 16(3):149-59.
72. Huet PM, Nagaoka MR, Desbiens G, Tarrab E, Brault A, Bralet MP, Bilodeau M. Sinusoidal endothelial cell and hepatocyte death following cold ischemia- warm reperfusion of the rat liver. Hepatology 2004; 39(4):1110-9.
73. Pinto VM, Allgayer MC, Pereira RA, Mello JRB, Breyer AS, Oliveira MEM. Avaliação do uso de Piper methysticum quanto ao desenvolvimento de hepatotoxicidade em ratos Wistar Revista Veterinária em Foco 2005; 3: 29-36. 74. Daemen M, Veer C, Denecker G, Heemskerk VH, Wolfs T, Clauss M,
Vandenabeele P, Buurman WA: Inhibition of apoptosis induced by ischemia- reperfusion prevents inflammation. J Clin Invest 1999; 104:541-549.
75. Daemen M, De Vries M, Van’t Veer C, Wolfs T, Buurman WA: Apoptosis and chemokine induction after renal ischemia-reperfusion. Transplantation 2001; 71:1007-1011.
56
apoptosis by distinct mechanisms. EMBO J 1999;18:189-198.
77. Sharma SK, Chapman D, Temsah R, Netticadan T, Brasil DP, Dhalla NS. Prevention of vascular apoptosis in myocardial infarction by Losartan. J Cardiovasc Pharmacol Ther 1999; 4: 77-84.
78. Arisawa EAL, Moraes E, Rocha RF, Almeida JD. Marcadores biológicos: PCNA e Ki-67. Breve revisão. Rev Fac Odontol 1999; 2: 54-60.
79. Yamada F, Saito T, Abe T, Tsuchiya T, Sato Y, Kenjo A, Kimura T, Gotoh M. Ischemic preconditioning enhances regenerative capacity of hepatocytes in long-term ischemically damaged rat livers. J Gastroenterol Hepatol. 2007; 22(11):1971-7.
80. Selzner N, Selzner M, Tian Y, Kadry Z, Clavien PA. Cold ischemia decreases liver regeneration after partial liver transplantation in the rat: A TNF-alpha/IL-6- dependent mechanism. Hepatology 2002; 36:812-8.
81. Selzner M, Camargo CA, Clavien PA.Ischemia impairs liver regeneration after major tissue loss in rodents: protective effects of interleukin-6. Hepatology 1999 ; 30(2):469-75.
82. Ledda-Columbano GM, Curto M, Piga R, Zedda AI, Menegazzi M, Sartori C, Shinozuka H, Bluethmann H, Poli V, Ciliberto G, Columbano A. In vivo hepatocyte proliferation is inducible through a TNF and IL-6-independent pathway. Oncogene 1998; 17(8):1039-44.
83. Montero EFS, Simões MJ, Abrahão MS, Ramalho CEB, Fagundes DJ, Salim CS. Efeito da N-acetilcisteína no pulmão após isquemia hepática em ratos. Acta Cir. Bras. 2002; 3:177-180.
84. Weinbroum AA, Rudick V, Ben-Abraham R, Karchevski E. N-acetyl-L-cysteine for preventing lung reperfusion injury after liver ischemia-reperfusion: a possible dual protective mechanism in a dose-response study. Transplantation. 2000 Mar 15;69(5):853-9.
85. Jin X, Wang L, Wu HS, Zhang L, Wang CY, Tian Y, Zhang JH. N-acetylcysteine inhibits activation of toll-like receptor 2 and 4 gene expression in the liver and lung after partial hepatic ischemia-reperfusion injury in mice. Hepatobiliary Pancreat Dis Int 2007; 6(3):284-9.
57
hepatoprotective and pro-proliferative effects of ischaemic preconditioning in mice. J Hepatol 2006; 45(1):20-7
87. Teoh N, Dela Pena A, Farrell G. Hepatic ischemic preconditioning in mice is associated with activation of NF-kappaB, p38 kinase, and cell cycle entry. Hepatology 2002; 36(1):94-102.
88. Rudiger HA, Clavien PA.Tumor necrosis factor alpha, but not Fas, mediates hepatocellular apoptosis in the murine ischemic liver. Gastroenterology 2002; 122(1):202-10.
89. Ozgüç H, Tokyay R, Kahveci N, Serdar Z, Gür ES, Korfali E. Oxothiazolidine Carboxylate provides protection against hepatocellular injury seen after porta hepatic occlusion (Pringle maneuver) under hypovolemic conditions. Word j Surg 2003; 27: 448-54.
90. Schauer RJ, Gerbes AL, Vonier D, Meissner H, Michl P, Leiderer R, Schildberg FW, Messmer K, Bilzer M. Glutathione protects the rat liver against reperfusion injury after prolonged warm ischemia. Ann Surg 2004; 239:220-31.
91. Hur GM, Ryu YS, Yun HY, Jeon BH, Kim YM, Seok JH, Lee JH. Hepatic Ischemia / reperfusion in rats induces iNOS gene transcription by activation of NF-kappaB. Biochem Biophys Res Commun. 1999; 261(3):917-22.
92. Kin H, Wang NP, Halkos ME, Kerendi F, Guyton RA, Zhao ZQ. Neutrophil depletion reduces myocardial apoptosis and attenuates NFkappaB activation/TNFalpha release after ischemia and reperfusion. J Surg Res. 2006; 135(1):170-8.
8. NORMAS ADOTADAS
INTERNATIONAL COMMITTEE OF MEDICAL JOURNAL EDITORS. Uniform requirements for manuscripts submitted to biomedical journals. Philadelphia: Ann Intern Med 1997; 126:36-47.
PÓS-GRADUAÇÃO EM CIRURGIA E EXPERIMENTAÇÃO – UNIFESP-EPM. Fagundes DJ. Normas para elaboração de relatório de pesquisa [edição eletrônica – disquete]. 2001 São Paulo (SP): UNIFESP-EPM-TOCE.
DeCS- Descritores em Ciências da Saúde. [serial online] 2001. Disponível em: URL: http://www.bireme.br
COLÉGIO BRASILEIRO DE EXPERIMENTAÇÃO ANIMAL (COBEA). Manual para técnicos em bioterismo. São Paulo (SP): Yellow Grafth; 1996.
59
9. ANEXOS
Tabela 1: Valores dos pesos dos ratos submetidos dos grupos IR, IPC, NAC submetidos a isquemia de 40 minutos e reperfusão de 24 horas e dos ratos do grupo SHAM.
Rato Sham IR NAC IPC
1 2 3 4 5 6 377 346 361 329 382 335 318 328 332 376 329 361 351 322 332 363 376 317 389 348 371 357 373 326 MÉDIA DP 355 21 341 22 343 23 361 22
Análise de variância para postos de Kruskal-Wallis
H critico =7,82 H calculado = 4.66 (p) Kruskal-Wallis = 0.19