Assessment of expressions of Bcl-XL, b-FGF, Bmp-2, Caspase-3, PDGFR-α, Smad1 and TGF-β1 genes in a rat model of lung ischemia/reperfusion

Document Type: Original Article

Authors

1 Dumlupınar University, Department of Physiology, Faculty of Medicine, Kütahya, Turkey

2 Gaziantep University, Department of Physiology, Faculty of Medicine, Gaziantep, Turkey

3 Fırat University, Department of Pathology, Faculty of Veterinary Science, Elazığ, Turkey

4 Gaziantep University, Department of Medical Biology, Faculty of Medicine, Gaziantep, Turkey

5 Gazi University, Department of Medical Genetic, Faculty of Medicine, Ankara, Turkey

Abstract

Objective(s):Ischemia is described as organs and tissues are destitute of oxygen due to decreased arterial or venous blood flow. Many mechanisms play role in cell death happened as a consequence of a new blood flow is needed for both cell regeneration and to clean toxic metabolites during ischemia and later. Lung damage induced by ischemia/reperfusion (I/R) is a frequent problem in lung transplantation. Apoptosis (programmed cell death) is known as cell suicide, and plays a key role in embryonic developmental and in maintain adult tissue’s life.
Materials and Methods:It is investigated expressions of Smad1, Bmp-2, Bcl-XL, b-FGF, Caspase-3, TGF-β1, PDGFR-α genes for molecular changes in lung tissues, after I/R is formed, in this study. For this, we included 40 Wistar albino rats to this study and divided 4 groups (n=10). The Groups were determined as Control (C), Group 1= 1 hr ischemia (I), Group 2= 1 hr ischemia+2 hr reperfusion (I+2R), Group 3= 1 hr ischemia+4 hr reperfusion (I+4R). Besides, molecular analysis and histopathologic examinations of tissues were performed, and the results were evaluated by normalization and statistics analysis.
Results: We have found a significant increase in expression of Bcl-XL (P=0.046) and Caspase-3 (P=0.026) genes of group 1, and it was not monitored any significant difference in Group 2 and Group 3. In all groups, the changes in b-FGF (P=0.087), Bmp-2 (P=0.457), TGF-β1 (P=0.201) and PDGFR-α (P=0.116) were not significant compared to control group. We did not see any mRNA expression of Smad1 gene in all groups include control.
Conclusion: These findings suggest that I/R injury may trigger apoptotic mechanism in lung.

Keywords


1. Torres RL, Beló-Klein A, Andrade CF, Cardoso PF. Effect of systemically administered low potassium dextran solution on oxidative stress in a rat model of lung ischemia. Interact Cardiovasc Thorac Surg 2009; 8:3-6.

2. Wilhelm J. Metabolic aspects of membrane lipid peroxidation. Acta Univ Carol Med Monogr 1990; 137:1-53

3. Ng CS, Wan S, Yim AP. Pulmonary ischaemia-reperfusion injury: role of apoptosis. Eur Respir J 2005; 25 :356-363.

4. Delbin MA, Antunes E, Zanesco A. Role of exercise training on pulmonary ischemia/reperfusion and inflammatory response. Rev Bras Cir Cardiovasc 2009; 24:552-561.

5. Ryter SW, Choi AM. Autophagy in the lung. Proc Am Thorac Soc 2010; 7:13-21.

6. den Hengst WA, den Hengst WA. Lung ischemia-reperfusion injury: a molecular and clinical view on a complex pathophysiological process. Am J Physiol Heart Circ Physiol 2010; 299:H1283-1299.

7. Hotchkiss RS, Strasser A, McDunn JE, Swanson PE. Cell death.  N Engl J Med 2009; 361:1570-1583.

8. Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972; 26:239-257.

9. Connolly PF, Jäger R, Fearnhead HO. New roles for old enzymes: killer caspases as the engine of cell behavior changes. Front Physiol 2014; 5:149.

10. Cheng EH, Wei MC, Weiler S, Flavell RA, Mak TW, Lindsten T, Korsmeyer SJ. BCL-2, BCL-X(L) sequester BH3 domain-only molecules preventing BAX- and BAK-mediated mitochondrial apoptosis. Mol Cell 2001; 8:705-711.

11. Chipuk JE, Moldoveanu T, Llambi F, Parsons MJ, Green DR. The BCL-2 family reunion. Mol Cell 2010; 37:299-310.

12. Gacche RN, Meshram RJ. Angiogenic factors as potential drug target: Efficacy and limitations of anti-angiogenic therapy. Biochim Biophys Acta 2014; 1846:161-179.

13. Li HH, Huo LJ, Gao ZY, Zhao F, Zeng JW. Regulation of scleral fibroblast differentiation by bone morphogenetic protein-2. Int J Ophthalmol 2014; 7:152-156.

14. Herhaus L, Sapkota GP. The emerging roles of deubiquitylating enzymes (DUBs) in the TGFbeta and BMP pathways. Cell Signal 2014; 26:2186-2192.

15. Schul D, Schmitt A, Regneri J, Schartl M, Wagner TU. Burst BMP triggered receptor kinase activity drives Smad1 mediated long-term target gene oscillation in C2C12 cells. PloS One 2013; 8:e59442.

16. Heldin CH. Targeting the PDGF signaling pathway in tumor treatment. Cell Commun Signal 2013; 11:97.

17. Takhtfooladi H, Takhtfooladi M, Moayer F, Mobarakeh S. Melatonin attenuates lung injury in a
hind limb ischemia-reperfusion rat model. Rev Port Pneumol 2015; 21:30-35.

18. Forgiarini LF, Forgiarini LA Jr, da Rosa DP, Silva MB, Mariano R, Paludo Ade O,et al. N-acetylcysteine administration confers lung protection in different phases of lung ischaemia-reperfusion injury. Interact Cardiovasc Thorac Surg 2014; 19:894-899.

19. Porter AG, Janicke RU. Emerging roles of caspase-3 in apoptosis. Cell Death  Differ 1999; 6:99-104.

20. Zhang Z, Shen H, Qin HD, Xu Y, Ma Mz, Bao L, Wang H. [Protective effect of N-acetylcysteine against pneumocyte apoptosis during ischemia/reperfusion injury of lung in rats]. Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue 2012; 24:111-115.

21. Zhang C, Guo Z, Lio H, Shi Y, Ge S. Influence of levosimendan postconditioning on apoptosis of rat lung cells in a model of ischemia-reperfusion injury. PloS One 2015; 10:e0114963.

22. Gao X, Cao Y, Staloch DA, Gonzales MA, Aronson JF. Bone morphogenetic protein signaling protects against cerulein-induced pancreatic fibrosis. PloS One 2014; 9:e89114.

23. Langenfeld EM, Kong Y, Langenfeld J. Bone morphogenetic protein 2 stimulation of tumor growth involves the activation of Smad-1/5. Oncogene 2006; 25:685-692.

24. Goumans MJ, Mummery C. Functional analysis of the TGFbeta receptor/Smad pathway through gene ablation in mice.  Int J Dev Biol 2000; 44:253-265.

25. Fu XB, Yang YH, Sun TZ, Gu XM, Jiang LX, Sun XQ, Sheng ZY. Effect of intestinal ischemia-reperfusion on expressions of endogenous basic fibroblast growth factor and transforming growth factor betain lung and its relation with lung repair. World J Gastroenterol 2000; 6:353-355

26. Florkiewicz RZ, Ahluwalia A, Sandor Z, Szabo S, Tarnawski AS. Gastric mucosal injury activates bFGF gene expression and triggers preferential translation of high molecular weight bFGF isoforms through CUG-initiated, non-canonical codons. Biochem Biophys Res Commun 2011; 409:494-499.

27. Sage E, Mercier O, Van den Eyden F, de Perrot M, Barlier-Mur AM, Dartevelle P, Eddahibi S. Endothelial cell apoptosis in chronically obstructed and reperfused pulmonary artery. Respir Res 2008; 9:19.