Ahmet TEKTEMUR, Ramazan Fazıl AKKOÇ, Elif ERDEM GÜZEL, Nalan KAYA TEKTEMUR

Adriamisin Uygulanan Sıçanların Böbrek Dokusunda Alfa Lipoik Asit’in Koruyucu Etkisinin DRP1 ile İlişkisi

Amaç: Çalışmada, adriamisin (ADR) uygulanan sıçanların böbrek dokularında meydana gelen değişiklikler üzerine alfa lipoik asit (ALA)’nın korucu etkisinin dinamin bağımlı protein 1 (DRP1) ile ilişkisinin incelenmesi amaçlanmıştır. Gereç ve Yöntem: Çalışmada, 28 adet Sprague-Dawley cinsi erkek sıçanlar kullanıldı. Deney hayvanları 4 eşit gruba ayrıldı. Kontrol grubuna deney süresi olan 28 gün boyunca hiçbir uygulama yapılmadı. ADR grubuna; 15 mg/kg ADR intraperitoneal (i.p) tek doz uygulandı. ADR+ALA grubuna 15 mg/kg i.p tek doz ADR verilmesinden sonra 50/mg/kg/gün dozunda ALA oral olarak gün aşırı verildi. ALA grubuna ise 50 mg/kg/gün dozunda ALA oral olarak gün aşırı uygulandı. Deney sonunda sıçanlar dekapite edildi. Dekapitasyonun ardından böbrek dokuları hızla çıkarıldı ve histolojik ve kantitatif RT-PCR analizleri için kullanıldı. Bulgular: ADR grubuna ait böbrek kesitlerinde inflamatuar hücre artışı, bowman boşluğunda artış, tübül lümeninde hiyalin birikimi gibi yapısal değişikliklere rastlandı. ALA uygulaması ile histopatolojik bulguların büyük bir çoğunluğunda azalma gözlendi. ADR grubunda DRP1 immünreaktivitesinin kontrol grubuna göre anlamlı artış gösterdiği, ADR+ALA grubunda ise ADR grubuna göre anlamlı derecede azalma olduğu belirlendi. Kontrol grubuna kıyasla ADR grubunda DRP1, BAX ve CASP3 mRNA düzeylerinde istatistiki açıdan anlamlı bir artış tespit edildi. ADR+ALA grubunda ise ADR grubuna kıyasla DRP1, BAX ve CASP3 mRNA düzeylerinde anlamlı bir azalma tespit edildi. Sonuç: Bu çalışma ADR’nin böbrek dokularında yapısal hasarla birlikte apoptozis ve DRP1 ekspresyon artışına yol açtığı, ALA’nın apoptozis ve DRP1 aktivitelerini düzenleyerek ADR’nin bu zararlı etkilerine karşı büyük oranda koruma sağladığı ortaya koyuldu.

The Relation of the Protective Effect of Alpha Lipoic Acid with DRP1 in the Kidney Tissue of Adriamycin Administered Rats

Objective: In this study, we aimed to investigate the association with DRP1 the protective effect of ALA on kidney tissue of the rats administered with adriamycin. Material and Method: Twenty eight Sprague-Dawley male rats were used in this study. Animals were divided to four groups. No administration was made to the control group during 28 days. A single dose of 15 mg / kg ADR was given intraperitoneally to the ADR group. After 15 mg / kg ADR was given to ADR + ALA group, 50 mg / kg ALA was given by oral gavage every other day. In the ALA group, 50 mg / kg ALA was administered orally every other day. At the end of the experiment, the rats were decapitated. Following decapitation, histological and quantitative RT-PCR analyzes were done. Results: Histopathological findings such as inflammatory cell increase, enlargement bowman distance were observed in the ADR group. ALA treatment was found to reduce the majority of histopathological findings.DRP1 immunoreactivity was significantly increased in the ADR group compared to the control group, whereas in the ADR+ALA group, DRP1 immunoreactivity was significantly lower than ADR group. A statistically significant increase was found in DRP1, BAX and CASP3 mRNA levels in the ADR group compared to the control group. A significant decrease was detected in DRP1, BAX and CASP3 mRNA levels in ADR + ALA group compared to the ADR group. Conclusion: This study revealed that ADR causes apoptosis and increases in DRP1 expression together with histopathological damages in kidney tissues, whereas ALA provides major protection against these effects of ADR by regulating apoptosis and DRP1 activities.

PDF

___

1. Gharanei M, Hussain A, Janneh O, Maddock HL. Doxorubicin induced myocardial injury is exacerbated following ischaemic stress via opening of the mitochondrial permeability transition pore. Toxicol Appl Pharmacol 2013; 149-56.
2. Yeh YC, Liu TJ, Wang LC et al. A standardized extract of Ginkgo biloba suppresses doxorubicininduced oxidative stress and p53-mediated mitochondrial apoptosis in rat testes. Br J
3. Ayla S, Seckin I, Tanriverdi G et al. Doxorubicin induced nephrotoxicity: protective effect of nicotinamide. Int J Cell Biol 2011; 1-9.
4. Miranda CJ, Makui H, Soares RJ et al. Hfe deficiency increases susceptibility to cardiotoxicity and exacerbates changes in ironmetabolism induced by doxorubicin. Blood 2003; 102: 2574-80.
5. Carvalho C, Santos RX, Cardoso S et al. Doxorubicin: the good, the bad and the ugly effect. Curr Med Chem 2009; 16: 3267-85.
6. Xin YF, You ZQ, Gao HY et al. Protective effect of lycium barbarum polysaccharides against doxorubicin-induced testicular toxicity in rats. Phytother Res 2012; 26: 716-21.
7. Lenaz G. The mitochondrial production of reactive oxygen species: mechanisms and implications in human pathology. IUBMB Life 2001; 52: 159- 64.
8. Perry HM, Huang L, Wilson WJ et al. Dynamin- Related Protein 1 Deficiency Promotes Recovery from AKI. Am Soc Nephrol 2018; 29: 194-206.
9. Wai T, Langer T. Mitochondrial dynamics and metabolic regulation. Trends Endocrinol Metab 2016; 27: 105-17.
10. Hu C, Huang Y, Li L. Drp1-dependent mitochondrial fission plays critical roles in physiological and pathological progresses in mammals. Int J Mol Sci 2017; 18: 144.
11. Pernas L, Scorrano L. Mito-Morphosis: mitochondrial fusion, fission, and cristae remodeling as key mediators of cellular function. Annu Rev Physiol 2016; 78: 505-31.
12. Yu T, Sheu SS, Robotham JL, Yoon Y. Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species. Cardiovasc Res 2008; 79: 341-51.
13. Wang JX, Li Q, Li PF. Apoptosis repressor with caspase recruitment domain contributes to chemotherapy resistance by abolishing mitochondrial fission mediated by dynamin-related protein-1. Cancer Res 2009; 69: 492-500.
14. Li Q, Wang JX, He YQ et al. MicroRNA-185 regulates chemotherapeutic sensitivity in gastric cancer by targeting apoptosis repressor with caspase recruitment domain. Cell Death Dis 2014; 5: 1197.
15. Reed LJ. From lipoic acid to multi-enzyme complexes. Protein Sci 1998; 7: 220-4.
16. Gorąca A, Huk-Kolega H, Piechota A, Kleniewska P, Ciejka E, Skibska B. Lipoic acid - biological activity and therapeutic potential. Pharmacol Rep 2011; 63: 849-58.
17. Prahalathan C, Selvakumar E, Varalakshmi P. Protective effect of lipoic acid on adriamycininduced testicular toxicity. Clinica Chimica Acta 2005; 360: 160-6.
18. Refaie MM. Amin EF. El-Tahawy NF, Abdelrahman AM. Possible Protective effect of diacerein on doxorubicin-induced nephrotoxicity in rats. J Toxicol 2016; 2016: 9507563.
19. El-Sayed EM, Mansour AM, El-Sawy WS. Alpha lipoic acid prevents doxorubicin-induced nephrotoxicity by mitigation of oxidative stress, inflammation, and apoptosis in rats. J Biochem Mol Toxicol 2017; 31. doi: 10.1002/jbt.21940.
20. Onishi H, Kuriyama K, Yamaguchi M et al. Concurrent two-dimensional radiotherapy and weekly docetaxel in the treatment of stage III nonsmall cell lung cancer: a good local response but no good survival due to radiation pneumonitis. Lung Cancer 2003; 40: 79-84.
21. Liu LL, Li QX, Xia L, Li J, Shao L. Differential effects of dihydropyridine calcium antagonists on doxorubicin induced nephrotoxicity in rats. Toxicology 2007; 231: 81-90.
22. Lee VWS, Harrıs DCH. Adriamycin nephropathy: A model of focal segmental glomerulosclerosis. Nephrology 2011; 16: 30-8.
23. Yagmurca M, Yasar Z, Bas O. Effects of quercetin on kidney injury induced by doxorubicin. Bratisl Med J 2015; 116: 486-9.
24. Karaman A, Fadillioglu E, Turkmen E, Tas E, Yilmaz Z. Protective effects of leflunomide against ischemia reperfusion injury of the rat liver. Pediatric Surgery International 2006; 22: 428-34.
25. Kagan V, Shvedova A, Serbinova E et al. Dihydrolipoic acid-A universal antioxidant both in the membrane and in the aqueous phase. Reduction of peroxyl, ascorbyl and chromanoxyl radicals. Biochem Pharmacol 1992; 44: 1637-49.
26. Malarkodi KP, Balachandar AV, Sivaprasad R, Varalakshmi P. Prophylactic effect of lipoic acid against adriamycin-ınduced peroxidative damages in rat kidney. Renal Faılure 2003; 25: 367-77. 27. Westermann B. Bioenergetic role of mitocho ndrial fusion and fission. Biochim Biophys Acta 2012; 1817: 1833-8.
28. Jheng HF, Tsai PJ, Guo SM et al. Mitochondrial fission contributes to mitochondrial dysfunction and insulin resistance in skeletal muscle. Mol Cell Biol 2012; 32: 309-19.
29. Ni Z, Tao L, Xiaohui X et al. Polydatin impairs mitochondria fitness and ameliorates podocyte injury by suppressing Drp1 expression. J Cell Physiol 2017; 232: 2776-87.
30. Yuan Y, Zhang A, Qi J et al. P53/Drp1-dependent mitochondrial fission mediates aldosteroneinduced podocyte injury and mitochondrial dysfunction. Am J Physiol Renal Physiol 2018; 314: 798-808.
31. Wang J, Feng C, He Y et al. Phosphorylation of apoptosis repressor with caspase recruitment domain by protein kinase CK2 contributes to chemotherapy resistance by inhibiting doxorubicin induced apoptosis. Oncotarget 2015; 6: 27700-13.
32. Perfettini JL, Roumier T, Kroemer G. Mitochondrial fusion and fission in the control of apoptosis. Trends Cell Biol 2005; 15: 179-83.
33. Loson OC, Song Z, Chen H, Chan DC. Fis1, Mff, MiD49, and MiD51 mediate Drp1 recruitment in mitochondrial fission. Mol Biol Cell 2013; 24: 659-67.
34. Xiaoa L, Xua X, Zhanga F et al. The mitochondria- targeted antioxidant MitoQ ameliorated tubular injury mediated by mitophagy in diabetic kidney disease via Nrf2/PINK1. Redox Biology 2017; 11: 297-311.
35. Su Z, Ye J, Qin Z, Dingb X. Protective effects of madecassoside against doxorubicin induced nephrotoxicity in vivo and in vitro. Scientific Reports 2015; 5: 1-14.