The Effect of Apilarnil on The Autophagia Against Lipolysaccarite-Based Sepsıs in Liver

Abstract

Sepsis, triggered by highly bacterial lipopolysaccharide (LPS) endotoxins, exhibits high morbidity and mortality despite medical advances. Damage to the liver occurs due to the production of highly reactive oxygen compounds (ROS) and the release of various proinflammatory cytokines. It is suggested that autophagy, which regulates inflammation and selectively destroys damaged mitochondria, suppresses apoptosis and provides a possible protective mechanism in the endotoxic liver. However, apilarnil, a bee product, is known to have high antioxidant activity and positive effects against various diseases thanks to its polyphenols. In this study, it is aimed to reveal the potential protective effect of apilarnil on the autophagy mechanism in the endotoxic liver model LPS-induced. 64 male Sprague dawley rats weighing 200-250; control, apilarnil treated groups (0.2, 0.4 and 0.8 g / kg), LPS (30 mg / kg) group and LPS + apilarnil treated groups (LPS + 0.2 g / kg, LPS + 0.4 g / kg and LPS + 0.8 g / kg) are randomly divided into eight groups. Beclin-1, LC3 and P62 proteins were analyzed immunohistochemically in order to determine the activity level of autophagy pathway in the liver tissues taken after the completion of the experiment protocol. The data obtained showed that Beclin-1 immunoreactivity decreased while LC3 and P62 expression increased in the tissues of the LPS group compared to the control group. When apilarnil was applied with LPS, it was determined that there was an increase in Beclin-1 level (p> 0.05) and a decrease in P62 levels (p <0.05) depending on the dose increase. Apilarnil increases the activity of the autophagy pathway and shows potential positive effects by providing a significant decrease on LC3 and P62 protein expression increased by LPS. However, the role of apilarnil in the autophagy pathway, which is a possible protector against LPS-induced sepsis, should be further investigated.

Keywords

• Beclin-1,LC3,P62,Apilarnil,LPS,Liver

References

1. References 1. Yan J, Li S, Li S. The role of the liver in sepsis. Int Rev Immunol. 2014;33(6):498-510. 2. Hotchkiss RS, Moldawer LL, Opal SM, Reinhart K, Turnbull IR, Vincent JL. Sepsis and septic shock. Nat Rev Dis Primers. 2016;2:16045. 3. Maldonado RF, Sa-Correia I, Valvano MA. Lipopolysaccharide modification in Gram-negative bacteria during chronic infection. FEMS Microbiol Rev. 2016;40(4):480-93. 4. Opal SM. Endotoxins and other sepsis triggers. Contrib Nephrol. 2010;167:14-24. 5. van der Poll T, van de Veerdonk FL, Scicluna BP, Netea MG. The immunopathology of sepsis and potential therapeutic targets. Nat Rev Immunol. 2017;17(7):407-20. 6. Cecconi M, Evans L, Levy M, Rhodes A. Sepsis and septic shock. Lancet. 2018;392(10141):75-87. 7. Sakr Y, Jaschinski U, Wittebole X, Szakmany T, Lipman J, Namendys-Silva SA, et al. Sepsis in Intensive Care Unit Patients: Worldwide Data From the Intensive Care over Nations Audit. Open Forum Infect Dis. 2018;5(12):ofy313. 8. Woznica EA, Inglot M, Woznica RK, Lysenko L. Liver dysfunction in sepsis. Adv Clin Exp Med. 2018;27(4):547-51. 9. Nesseler N, Launey Y, Aninat C, Morel F, Malledant Y, Seguin P. Clinical review: The liver in sepsis. Crit Care. 2012;16(5):235. 10. Srivastava B, Gimson A. Hepatic changes in systemic infection. Best Pract Res Clin Gastroenterol. 2013;27(4):485-95. 11. Marshall JC. New translational research provides insights into liver dysfunction in sepsis. PLoS Med. 2012;9(11):e1001341. 12. De Nardo D. Toll-like receptors: Activation, signalling and transcriptional modulation. Cytokine. 2015;74(2):181-9. 13. Kolios G, Valatas V, Manousou P, Xidakis C, Notas G, Kouroumalis E. Nitric oxide and MCP-1 regulation in LPS activated rat Kupffer cells. Mol Cell Biochem. 2008;319(1-2):91-8. 14. Wang D, Yin Y, Yao Y. Advances in sepsis-associated liver dysfunction. Burns Trauma. 2014;2(3):97-105. 15. Xu Y, Jagannath C, Liu XD, Sharafkhaneh A, Kolodziejska KE, Eissa NT. Toll-like receptor 4 is a sensor for autophagy associated with innate immunity. Immunity. 2007;27(1):135-44. 16. Fujita K, Maeda D, Xiao Q, Srinivasula SM. Nrf2-mediated induction of p62 controls Toll-like receptor-4-driven aggresome-like induced structure formation and autophagic degradation. Proc Natl Acad Sci U S A. 2011;108(4):1427-32. 17. Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A, et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1):1-222. 18. Yu L, Chen Y, Tooze SA. Autophagy pathway: Cellular and molecular mechanisms. Autophagy. 2018;14(2):207-15. 19. Hu W, Chan H, Lu L, Wong KT, Wong SH, Li MX, et al. Autophagy in intracellular bacterial infection. Semin Cell Dev Biol. 2020;101:41-50. 20. Feng Y, Liu B, Zheng X, Chen L, Chen W, Fang Z. The protective role of autophagy in sepsis. Microb Pathog. 2019;131:106-11. 21. Yin X, Xin H, Mao S, Wu G, Guo L. The Role of Autophagy in Sepsis: Protection and Injury to Organs. Front Physiol. 2019;10:1071. 22. Oami T, Watanabe E, Hatano M, Sunahara S, Fujimura L, Sakamoto A, et al. Suppression of T Cell Autophagy Results in Decreased Viability and Function of T Cells Through Accelerated Apoptosis in a Murine Sepsis Model. Crit Care Med. 2017;45(1):e77-e85. 23. Ding WX, Yin XM. Mitophagy: mechanisms, pathophysiological roles, and analysis. Biol Chem. 2012;393(7):547-64. 24. Chang AL, Ulrich A, Suliman HB, Piantadosi CA. Redox regulation of mitophagy in the lung during murine Staphylococcus aureus sepsis. Free Radic Biol Med. 2015;78:179-89. 25. Baechler BL, Bloemberg D, Quadrilatero J. Mitophagy regulates mitochondrial network signaling, oxidative stress, and apoptosis during myoblast differentiation. Autophagy. 2019;15(9):1606-19. 26. Aki T, Unuma K, Uemura K. Emerging roles of mitochondria and autophagy in liver injury during sepsis. Cell Stress. 2017;1(2):79-89. 27. Akçiçek E, Yücel B, . Apiterapi’de Apilarnil. Arı Ürünleri ve Sağlık (Apiterapi). İzmir: Sidas Yayınevi; 2015. 28. Isİdorov VA, Bakier S, Stocki M. GC-MS investigation of the chemical composition of honeybee drone and queen larva homogenate. J Apic Res. 2016;60(1):111-20. 29. Silici S. Honeybee Products and Apitherapy. . Turkish Journal of Agriculture-Food Science and Technology. 2019 7(9):1249-62. 30. Silici S. Chemical Content and Bioactive Properties of Drone Larvae (Apilarnil). Mellifera. 2019 19(2):14-22. 31. Meda A, Lamien CE, Millogo J, Romito M, Nacoulma OG. Therapeutic uses of honey and honeybee larvae in central Burkina Faso. J Ethnopharmacol. 2004;95(1):103-7. 32. Doganyigit Z, Kup FO, Silici S, Deniz K, Yakan B, Atayoglu T. Protective effects of propolis on female rats' histopathological, biochemical and genotoxic changes during LPS induced endotoxemia. Phytomedicine. 2013;20(7):632-9. 33. Kanbur M, Eraslan G, Beyaz L, Silici S, Liman BC, Altinordulu S, et al. The effects of royal jelly on liver damage induced by paracetamol in mice. Exp Toxicol Pathol. 2009;61(2):123-32. 34. Doganyigit Z, Okan A, Kaymak E, Pandir D, Silici S. Investigation of protective effects of apilarnil against lipopolysaccharide induced liver injury in rats via TLR 4/ HMGB-1/ NF-kappaB pathway. Biomed Pharmacother. 2020;125:109967. 35. Watanabe E, Muenzer JT, Hawkins WG, Davis CG, Dixon DJ, McDunn JE, et al. Sepsis induces extensive autophagic vacuolization in hepatocytes: a clinical and laboratory-based study. Lab Invest. 2009;89(5):549-61. 36. Sun Q, Gao W, Loughran P, Shapiro R, Fan J, Billiar TR, et al. Caspase 1 activation is protective against hepatocyte cell death by up-regulating beclin 1 protein and mitochondrial autophagy in the setting of redox stress. J Biol Chem. 2013;288(22):15947-58. 37. Kroemer G, Marino G, Levine B. Autophagy and the integrated stress response. Mol Cell. 2010;40(2):280-93. 38. Levine B, Kroemer G. Autophagy in the pathogenesis of disease. Cell. 2008;132(1):27-42. 39. Betin VM, Lane JD. Caspase cleavage of Atg4D stimulates GABARAP-L1 processing and triggers mitochondrial targeting and apoptosis. J Cell Sci. 2009;122(Pt 14):2554-66. 40. Carchman EH, Rao J, Loughran PA, Rosengart MR, Zuckerbraun BS. Heme oxygenase-1-mediated autophagy protects against hepatocyte cell death and hepatic injury from infection/sepsis in mice. Hepatology. 2011;53(6):2053-62. 41. Puissant A, Fenouille N, Auberger P. When autophagy meets cancer through p62/SQSTM1. Am J Cancer Res. 2012;2(4):397-413. 42. Lazova R, Camp RL, Klump V, Siddiqui SF, Amaravadi RK, Pawelek JM. Punctate LC3B expression is a common feature of solid tumors and associated with proliferation, metastasis, and poor outcome. Clin Cancer Res. 2012;18(2):370-9. 43. Komatsu M, Kurokawa H, Waguri S, Taguchi K, Kobayashi A, Ichimura Y, et al. The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1. Nat Cell Biol. 2010;12(3):213-23. 44. Chen C, Deng M, Sun Q, Loughran P, Billiar TR, Scott MJ. Lipopolysaccharide stimulates p62-dependent autophagy-like aggregate clearance in hepatocytes. Biomed Res Int. 2014;2014:267350. 45. Waltz P, Carchman EH, Young AC, Rao J, Rosengart MR, Kaczorowski D, et al. Lipopolysaccaride induces autophagic signaling in macrophages via a TLR4, heme oxygenase-1 dependent pathway. Autophagy. 2011;7(3):315-20. 46. Carchman EH, Whelan S, Loughran P, Mollen K, Stratamirovic S, Shiva S, et al. Experimental sepsis-induced mitochondrial biogenesis is dependent on autophagy, TLR4, and TLR9 signaling in liver. FASEB J. 2013;27(12):4703-11.