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Protective Effects of Apocynin On Cecal Ligation and Puncture-Induced Lung and Renal Injuries

Yıl 2020, Cilt: 8 Sayı: 1, 7 - 11, 30.04.2020

Öz

Aim: The possible beneficial properties of Apocynin against cecal ligation and puncture (CLP) induced lung and renal injuries were investigated in rats.
Materials and Methods: 32 Wistar Albino male rats were randomized as: group I (sham), group II (CLP), group III (CLP+Apocynin 20 mg/kg), and group IV (CLP+Apocynin 50 mg/kg). CLP process was carried out under anesthesia conditions. Apocynin was administered intraperitoneally prior to the CLP model. The lung and renal tissues were excised following the experiment. Biochemical analyzes were done.
Results and Conclusion: TNF-α, OSI, IL-1β, TOS, MDA levels and MPO activity elevated but SOD and TAS values declined in CLP group compared to sham group. On the other hand, SOD and TAS levels increased while MPO activity, TNF-α, IL-1β, TOS, OSI and MDA levels declined due to Apocynin treatments. As a conclusion, Apocynin is an effective agent against CLP-induced lung and renal injuries.

Kaynakça

  • 1. Hattori, Y., et al., Recent advances in the pathophysiology and molecular basis of sepsis-associated organ dysfunction: Novel therapeutic implications and challenges. Pharmacol Ther, 2017. 177: p. 56-66.
  • 2. Dellinger, R.P., et al., Surviving Sepsis Campaign: international guidelines for management of severe sepsis and septic shock, 2012. Intensive Care Med, 2013. 39(2): p. 165-228.
  • 3. Fu, J., et al., Sodium Butyrate Ameliorates Intestinal Injury and Improves Survival in a Rat Model of Cecal Ligation and Puncture-Induced Sepsis. Inflammation, 2019. 42(4): p. 1276-1286.
  • 4. Andrews, P., et al., Year in review in intensive care medicine. 2005. I. Acute respiratory failure and acute lung injury, ventilation, hemodynamics, education, renal failure. Intensive Care Med, 2006. 32(2): p. 207-216.
  • 5. Zimmerman, J.J., et al., Incidence and outcomes of pediatric acute lung injury. Pediatrics, 2009. 124(1): p. 87-95.
  • 6. Peters, E., et al., A worldwide multicentre evaluation of the influence of deterioration or improvement of acute kidney injury on clinical outcome in critically ill patients with and without sepsis at ICU admission: results from The Intensive Care Over Nations audit. Crit Care, 2018. 22(1): p. 188.
  • 7. Wang, J., et al., Interleukin-10 secreted by mesenchymal stem cells attenuates acute liver failure through inhibiting pyroptosis. Hepatol Res, 2018. 48(3): p. E194-e202.
  • 8. Chousterman, B.G., F.K. Swirski, and G.F. Weber, Cytokine storm and sepsis disease pathogenesis. Semin Immunopathol, 2017. 39(5): p. 517-528.
  • 9. Sun, G., et al., Esculentoside A ameliorates cecal ligation and puncture-induced acute kidney injury in rats. Exp Anim, 2017. 66(4): p. 303-312.
  • 10. Polat, B., et al., The protective effect of amiodarone in lung tissue of cecal ligation and puncture-induced septic rats: a perspective from inflammatory cytokine release and oxidative stress. Naunyn Schmiedebergs Arch Pharmacol, 2013. 386(7): p. 635-43.
  • 11. Siebenlist, U., G. Franzoso, and K. Brown, Structure, regulation and function of NF-kappa B. Annu Rev Cell Biol, 1994. 10: p. 405-55.
  • 12. Coskun, A.K., et al., The effects of montelukast on antioxidant enzymes and proinflammatory cytokines on the heart, liver, lungs, and kidneys in a rat model of cecal ligation and puncture-induced sepsis. ScientificWorldJournal, 2011. 11: p. 1341-56.
  • 13. Stefanska, J. and R. Pawliczak, Apocynin: molecular aptitudes. Mediators Inflamm, 2008. 2008: p. 106507.
  • 14. Stolk, J., et al., Characteristics of the inhibition of NADPH oxidase activation in neutrophils by apocynin, a methoxy-substituted catechol. Am J Respir Cell Mol Biol, 1994. 11(1): p. 95-102.
  • 15. Ximenes, V.F., et al., The oxidation of apocynin catalyzed by myeloperoxidase: proposal for NADPH oxidase inhibition. Arch Biochem Biophys, 2007. 457(2): p. 134-41.
  • 16. Ohkawa, H., N. Ohishi, and K. Yagi, Assay for Lipid Peroxides in Animal-Tissues by Thiobarbituric Acid Reaction. Analytical Biochemistry, 1979. 95(2): p. 351-358.
  • 17. Bradley, P.P., et al., Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Invest Dermatol, 1982. 78(3): p. 206-9.
  • 18. Sun, Y., L.W. Oberley, and Y. Li, A Simple Method for Clinical Assay of Superoxide-Dismutase. Clinical Chemistry, 1988. 34(3): p. 497-500.
  • 19. Harasstani, O.A., C.L. Tham, and D.A. Israf, Kaempferol and Chrysin Synergies to Improve Septic Mice Survival. Molecules, 2017. 22(1).
  • 20. Kim, W.Y. and S.B. Hong, Sepsis and Acute Respiratory Distress Syndrome: Recent Update. Tuberc Respir Dis (Seoul), 2016. 79(2): p. 53-7.
  • 21. Seeley, E.J., M.A. Matthay, and P.J. Wolters, Inflection points in sepsis biology: from local defense to systemic organ injury. Am J Physiol Lung Cell Mol Physiol, 2012. 303(5): p. L355-63.
  • 22. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980-2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet, 2017. 390(10100): p. 1151-1210.
  • 23. Fani, F., et al., Recent advances in the pathogenetic mechanisms of sepsis-associated acute kidney injury. J Nephrol, 2018. 31(3): p. 351-359.
  • 24. Hotchkiss, R.S., et al., Sepsis and septic shock. Nat Rev Dis Primers, 2016. 2: p. 16045.
  • 25. Güzel Erdoğan, D., Tanyeli A., Inhibition of NADPH oxidase attenuates sepsis induced acute lung oxidative damage in rats. Journal of Cellular Neuroscience & Oxidative Stress 2018. 10(2).
  • 26. Tanyeli A., Güzel Erdoğan D., Alliin mitigates Cecal Ligation Puncture (CLP)-induced lung injury through antioxidant and antiinflammatory effects. Turhish Journal of Sciences, 2019. 4(2): p. 46-59.
  • 27. Tanyeli A., Güzel E., An Investigation into the Biochemical Effects of Barbaloin on Renal Tissue in Cecal Ligation and Puncture-Induced Polymicrobial Sepsis Model in Rats. SCIE, 2019. 30(4): p. 285-289.
  • 28. Tanyeli A., et al., Investigation of biochemical and histopathological effects of tarantula cubensis D6 on lung tissue in cecal ligation and puncture-induced polymicrobial sepsis model in rats. 2019. 8: p. 644.
  • 29. Dejager, L., et al., Cecal ligation and puncture: the gold standard model for polymicrobial sepsis? Trends Microbiol, 2011. 19(4): p. 198-208.
  • 30. van der Poll, T., et al., The immunopathology of sepsis and potential therapeutic targets. Nat Rev Immunol, 2017. 17(7): p. 407-420.
  • 31. Rudd, K.E., et al., The global burden of sepsis: barriers and potential solutions. Crit Care, 2018. 22(1): p. 232.
  • 32. Koksal, G.M., et al., Correlation of plasma and tissue oxidative stresses in intra-abdominal sepsis. J Surg Res, 2004. 122(2): p. 180-3.
  • 33. Bone, R.C., C.J. Grodzin, and R.A. Balk, Sepsis: a new hypothesis for pathogenesis of the disease process. Chest, 1997. 112(1): p. 235-43.
  • 34. Hotchkiss, R.S., et al., The sepsis seesaw: tilting toward immunosuppression. Nat Med, 2009. 15(5): p. 496-7.
  • 35. Smith, J.A., P.R. Mayeux, and R.G. Schnellmann, Delayed Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Inhibition by Trametinib Attenuates Systemic Inflammatory Responses and Multiple Organ Injury in Murine Sepsis. Crit Care Med, 2016. 44(8): p. e711-20.
  • 36. Michels, M., et al., The role of microglia activation in the development of sepsis-induced long-term cognitive impairment. Brain Behav Immun, 2015. 43: p. 54-9.
  • 37. Mina, F., et al., Il1-β involvement in cognitive impairment after sepsis. Mol Neurobiol, 2014. 49(2): p. 1069-76.
  • 38. Mullane, K.M., R. Kraemer, and B. Smith, Myeloperoxidase activity as a quantitative assessment of neutrophil infiltration into ischemic myocardium. J Pharmacol Methods, 1985. 14(3): p. 157-67.
  • 39. Topdağı Ö, T.A., Ekinci Akdemir FN, Güzel Erdoğan D, Güler MC, Eraslan E. , The Effects of Higenamine on Testicular Damage Injured by Ischemia Reperfusion: A Biochemical Study. Turkish Journal of Science, 2019. 4(2): p. 92-99.
  • 40. Li, S., et al., Effects of adiponectin on mortality and its mechanism in a sepsis mouse model. J Invest Surg, 2012. 25(4): p. 214-9.
  • 41. Till, G.O., et al., Lipid peroxidation and acute lung injury after thermal trauma to skin. Evidence of a role for hydroxyl radical. Am J Pathol, 1985. 119(3): p. 376-84.
  • 42. Wang, Y., et al., Alleviation of Acute Lung Injury in Rats with Sepsis by Resveratrol via the Phosphatidylinositol 3-Kinase/Nuclear Factor-Erythroid 2 Related Factor 2/Heme Oxygenase-1 (PI3K/Nrf2/HO-1) Pathway. Med Sci Monit, 2018. 24: p. 3604-3611.
  • 43. Peng, Q.Y., et al., Blocking Cyclic Adenosine Diphosphate Ribose-mediated Calcium Overload Attenuates Sepsis-induced Acute Lung Injury in Rats. Chin Med J (Engl), 2016. 129(14): p. 1725-30.
  • 44. Fang, Y.Z., S. Yang, and G. Wu, Free radicals, antioxidants, and nutrition. Nutrition, 2002. 18(10): p. 872-9.
  • 45. Erel, O., A new automated colorimetric method for measuring total oxidant status. Clin Biochem, 2005. 38(12): p. 1103-11.
  • 46. Erel, O., A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem, 2004. 37(4): p. 277-85.
  • 47. Damas, P., et al., Sepsis and serum cytokine concentrations. Crit Care Med, 1997. 25(3): p. 405-12.
  • 48. Razavi-Azarkhiavi, K., et al., Silymarin alleviates bleomycin-induced pulmonary toxicity and lipid peroxidation in mice. Pharm Biol, 2014. 52(10): p. 1267-71.
  • 49. Touyz, R.M., Apocynin, NADPH oxidase, and vascular cells: a complex matter. Hypertension, 2008. 51(2): p. 172-4.
  • 50. Petrônio, M.S., et al., Apocynin: chemical and biophysical properties of a NADPH oxidase inhibitor. Molecules, 2013. 18(3): p. 2821-2839.
  • 51. Rahman, M.M., et al., Apocynin prevented inflammation and oxidative stress in carbon tetra chloride induced hepatic dysfunction in rats. Biomedicine & Pharmacotherapy, 2017. 92: p. 421-428.

Apokininin Çekal Ligasyon ve Ponksiyon ile İndüklenen Akciğer ve Böbrek Hasarına Karşı Koruyucu Etkileri

Yıl 2020, Cilt: 8 Sayı: 1, 7 - 11, 30.04.2020

Öz

Amaç: Sıçanlarda Apokinin’in, çekal ligasyon ve ponksiyona (CLP) bağlı akciğer ve böbrek hasarına karşı olası yararlı özellikleri araştırıldı.
Gereç ve Yöntem: 32 Wistar Albino erkek sıçan randomize olarak grup I (sham), grup II (CLP), grup III (CLP+Apokinin 20 mg/kg) ve grup IV (CLP+Apokinin 50 mg/kg)
olarak düzenlendi. CLP işlemi anestezi koşulları altında gerçekleştirildi. Apokinin, CLP modelinden hemen önce intraperitonal olarak uygulandı. Akciğer ve böbrek dokuları
deneyin ardından eksize edildi. Biyokimyasal analizler yapıldı.
Bulgular ve Sonuç: Sham grubuna kıyasla CLP grubunda TNF-a, OSI, IL-1B, TOS, MDA düzeyleri ve MPO aktivitesi yükselirken SOD ve TAS değerleri azaldı. Öte yandan,
Apokinin uygulaması sonucunda MPO aktivitesi, TNF-α, IL-1β, TOS, OSI ve MDA seviyeleri düşerken SOD ve TAS düzeyleri artmıştır. Sonuç olarak, Apokinin CLP kaynaklı
akciğer ve böbrek hasarına karşı etkili bir ajandır. 

Kaynakça

  • 1. Hattori, Y., et al., Recent advances in the pathophysiology and molecular basis of sepsis-associated organ dysfunction: Novel therapeutic implications and challenges. Pharmacol Ther, 2017. 177: p. 56-66.
  • 2. Dellinger, R.P., et al., Surviving Sepsis Campaign: international guidelines for management of severe sepsis and septic shock, 2012. Intensive Care Med, 2013. 39(2): p. 165-228.
  • 3. Fu, J., et al., Sodium Butyrate Ameliorates Intestinal Injury and Improves Survival in a Rat Model of Cecal Ligation and Puncture-Induced Sepsis. Inflammation, 2019. 42(4): p. 1276-1286.
  • 4. Andrews, P., et al., Year in review in intensive care medicine. 2005. I. Acute respiratory failure and acute lung injury, ventilation, hemodynamics, education, renal failure. Intensive Care Med, 2006. 32(2): p. 207-216.
  • 5. Zimmerman, J.J., et al., Incidence and outcomes of pediatric acute lung injury. Pediatrics, 2009. 124(1): p. 87-95.
  • 6. Peters, E., et al., A worldwide multicentre evaluation of the influence of deterioration or improvement of acute kidney injury on clinical outcome in critically ill patients with and without sepsis at ICU admission: results from The Intensive Care Over Nations audit. Crit Care, 2018. 22(1): p. 188.
  • 7. Wang, J., et al., Interleukin-10 secreted by mesenchymal stem cells attenuates acute liver failure through inhibiting pyroptosis. Hepatol Res, 2018. 48(3): p. E194-e202.
  • 8. Chousterman, B.G., F.K. Swirski, and G.F. Weber, Cytokine storm and sepsis disease pathogenesis. Semin Immunopathol, 2017. 39(5): p. 517-528.
  • 9. Sun, G., et al., Esculentoside A ameliorates cecal ligation and puncture-induced acute kidney injury in rats. Exp Anim, 2017. 66(4): p. 303-312.
  • 10. Polat, B., et al., The protective effect of amiodarone in lung tissue of cecal ligation and puncture-induced septic rats: a perspective from inflammatory cytokine release and oxidative stress. Naunyn Schmiedebergs Arch Pharmacol, 2013. 386(7): p. 635-43.
  • 11. Siebenlist, U., G. Franzoso, and K. Brown, Structure, regulation and function of NF-kappa B. Annu Rev Cell Biol, 1994. 10: p. 405-55.
  • 12. Coskun, A.K., et al., The effects of montelukast on antioxidant enzymes and proinflammatory cytokines on the heart, liver, lungs, and kidneys in a rat model of cecal ligation and puncture-induced sepsis. ScientificWorldJournal, 2011. 11: p. 1341-56.
  • 13. Stefanska, J. and R. Pawliczak, Apocynin: molecular aptitudes. Mediators Inflamm, 2008. 2008: p. 106507.
  • 14. Stolk, J., et al., Characteristics of the inhibition of NADPH oxidase activation in neutrophils by apocynin, a methoxy-substituted catechol. Am J Respir Cell Mol Biol, 1994. 11(1): p. 95-102.
  • 15. Ximenes, V.F., et al., The oxidation of apocynin catalyzed by myeloperoxidase: proposal for NADPH oxidase inhibition. Arch Biochem Biophys, 2007. 457(2): p. 134-41.
  • 16. Ohkawa, H., N. Ohishi, and K. Yagi, Assay for Lipid Peroxides in Animal-Tissues by Thiobarbituric Acid Reaction. Analytical Biochemistry, 1979. 95(2): p. 351-358.
  • 17. Bradley, P.P., et al., Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Invest Dermatol, 1982. 78(3): p. 206-9.
  • 18. Sun, Y., L.W. Oberley, and Y. Li, A Simple Method for Clinical Assay of Superoxide-Dismutase. Clinical Chemistry, 1988. 34(3): p. 497-500.
  • 19. Harasstani, O.A., C.L. Tham, and D.A. Israf, Kaempferol and Chrysin Synergies to Improve Septic Mice Survival. Molecules, 2017. 22(1).
  • 20. Kim, W.Y. and S.B. Hong, Sepsis and Acute Respiratory Distress Syndrome: Recent Update. Tuberc Respir Dis (Seoul), 2016. 79(2): p. 53-7.
  • 21. Seeley, E.J., M.A. Matthay, and P.J. Wolters, Inflection points in sepsis biology: from local defense to systemic organ injury. Am J Physiol Lung Cell Mol Physiol, 2012. 303(5): p. L355-63.
  • 22. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980-2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet, 2017. 390(10100): p. 1151-1210.
  • 23. Fani, F., et al., Recent advances in the pathogenetic mechanisms of sepsis-associated acute kidney injury. J Nephrol, 2018. 31(3): p. 351-359.
  • 24. Hotchkiss, R.S., et al., Sepsis and septic shock. Nat Rev Dis Primers, 2016. 2: p. 16045.
  • 25. Güzel Erdoğan, D., Tanyeli A., Inhibition of NADPH oxidase attenuates sepsis induced acute lung oxidative damage in rats. Journal of Cellular Neuroscience & Oxidative Stress 2018. 10(2).
  • 26. Tanyeli A., Güzel Erdoğan D., Alliin mitigates Cecal Ligation Puncture (CLP)-induced lung injury through antioxidant and antiinflammatory effects. Turhish Journal of Sciences, 2019. 4(2): p. 46-59.
  • 27. Tanyeli A., Güzel E., An Investigation into the Biochemical Effects of Barbaloin on Renal Tissue in Cecal Ligation and Puncture-Induced Polymicrobial Sepsis Model in Rats. SCIE, 2019. 30(4): p. 285-289.
  • 28. Tanyeli A., et al., Investigation of biochemical and histopathological effects of tarantula cubensis D6 on lung tissue in cecal ligation and puncture-induced polymicrobial sepsis model in rats. 2019. 8: p. 644.
  • 29. Dejager, L., et al., Cecal ligation and puncture: the gold standard model for polymicrobial sepsis? Trends Microbiol, 2011. 19(4): p. 198-208.
  • 30. van der Poll, T., et al., The immunopathology of sepsis and potential therapeutic targets. Nat Rev Immunol, 2017. 17(7): p. 407-420.
  • 31. Rudd, K.E., et al., The global burden of sepsis: barriers and potential solutions. Crit Care, 2018. 22(1): p. 232.
  • 32. Koksal, G.M., et al., Correlation of plasma and tissue oxidative stresses in intra-abdominal sepsis. J Surg Res, 2004. 122(2): p. 180-3.
  • 33. Bone, R.C., C.J. Grodzin, and R.A. Balk, Sepsis: a new hypothesis for pathogenesis of the disease process. Chest, 1997. 112(1): p. 235-43.
  • 34. Hotchkiss, R.S., et al., The sepsis seesaw: tilting toward immunosuppression. Nat Med, 2009. 15(5): p. 496-7.
  • 35. Smith, J.A., P.R. Mayeux, and R.G. Schnellmann, Delayed Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Inhibition by Trametinib Attenuates Systemic Inflammatory Responses and Multiple Organ Injury in Murine Sepsis. Crit Care Med, 2016. 44(8): p. e711-20.
  • 36. Michels, M., et al., The role of microglia activation in the development of sepsis-induced long-term cognitive impairment. Brain Behav Immun, 2015. 43: p. 54-9.
  • 37. Mina, F., et al., Il1-β involvement in cognitive impairment after sepsis. Mol Neurobiol, 2014. 49(2): p. 1069-76.
  • 38. Mullane, K.M., R. Kraemer, and B. Smith, Myeloperoxidase activity as a quantitative assessment of neutrophil infiltration into ischemic myocardium. J Pharmacol Methods, 1985. 14(3): p. 157-67.
  • 39. Topdağı Ö, T.A., Ekinci Akdemir FN, Güzel Erdoğan D, Güler MC, Eraslan E. , The Effects of Higenamine on Testicular Damage Injured by Ischemia Reperfusion: A Biochemical Study. Turkish Journal of Science, 2019. 4(2): p. 92-99.
  • 40. Li, S., et al., Effects of adiponectin on mortality and its mechanism in a sepsis mouse model. J Invest Surg, 2012. 25(4): p. 214-9.
  • 41. Till, G.O., et al., Lipid peroxidation and acute lung injury after thermal trauma to skin. Evidence of a role for hydroxyl radical. Am J Pathol, 1985. 119(3): p. 376-84.
  • 42. Wang, Y., et al., Alleviation of Acute Lung Injury in Rats with Sepsis by Resveratrol via the Phosphatidylinositol 3-Kinase/Nuclear Factor-Erythroid 2 Related Factor 2/Heme Oxygenase-1 (PI3K/Nrf2/HO-1) Pathway. Med Sci Monit, 2018. 24: p. 3604-3611.
  • 43. Peng, Q.Y., et al., Blocking Cyclic Adenosine Diphosphate Ribose-mediated Calcium Overload Attenuates Sepsis-induced Acute Lung Injury in Rats. Chin Med J (Engl), 2016. 129(14): p. 1725-30.
  • 44. Fang, Y.Z., S. Yang, and G. Wu, Free radicals, antioxidants, and nutrition. Nutrition, 2002. 18(10): p. 872-9.
  • 45. Erel, O., A new automated colorimetric method for measuring total oxidant status. Clin Biochem, 2005. 38(12): p. 1103-11.
  • 46. Erel, O., A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem, 2004. 37(4): p. 277-85.
  • 47. Damas, P., et al., Sepsis and serum cytokine concentrations. Crit Care Med, 1997. 25(3): p. 405-12.
  • 48. Razavi-Azarkhiavi, K., et al., Silymarin alleviates bleomycin-induced pulmonary toxicity and lipid peroxidation in mice. Pharm Biol, 2014. 52(10): p. 1267-71.
  • 49. Touyz, R.M., Apocynin, NADPH oxidase, and vascular cells: a complex matter. Hypertension, 2008. 51(2): p. 172-4.
  • 50. Petrônio, M.S., et al., Apocynin: chemical and biophysical properties of a NADPH oxidase inhibitor. Molecules, 2013. 18(3): p. 2821-2839.
  • 51. Rahman, M.M., et al., Apocynin prevented inflammation and oxidative stress in carbon tetra chloride induced hepatic dysfunction in rats. Biomedicine & Pharmacotherapy, 2017. 92: p. 421-428.
Toplam 51 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Sağlık Kurumları Yönetimi
Bölüm Orjinal Araştırma Makaleleri
Yazarlar

Ersen Eraslan 0000-0003-2424-2269

Derya Güzel 0000-0002-7618-5043

Songül Doğanay 0000-0002-1730-1331

Ayhan Tanyeli

Mustafa Can Güler 0000-0001-8588-1035

Yayımlanma Tarihi 30 Nisan 2020
Kabul Tarihi 21 Temmuz 2020
Yayımlandığı Sayı Yıl 2020 Cilt: 8 Sayı: 1

Kaynak Göster

Vancouver Eraslan E, Güzel D, Doğanay S, Tanyeli A, Güler MC. Protective Effects of Apocynin On Cecal Ligation and Puncture-Induced Lung and Renal Injuries. pediatr pract res. 2020;8(1):7-11.