Sıçanlarda Sisplatin Kaynaklı Hepatorenal Toksisiteye Karşı 2-Aminoetoksidifenil Boratın Koruyucu Etkileri

Öz

Sisplatin, solid tümörlerin tedavisinde yaygın olarak kullanılan platin bazlı bir kemoterapötiktir; ancak klinik kullanımı, oksidatif stres ve hücre içi Ca²⁺ homeostazındaki bozulma ile ilişkili doz bağımlı hepatotoksisite ve nefrotoksisite nedeniyle sınırlanmaktadır. Bor içeren bir IP₃ reseptör antagonisti olan 2-Aminoetoksidifenil borat (2-APB), kalsiyum sinyalizasyonunun modülasyonu yoluyla sitoprotektif etki gösterebilir. Bu çalışmada, 2-APB’nin sisplatin kaynaklı hepatorenal toksisiteye karşı profilaktik etkisi biyokimyasal ve histopatolojik parametreler kullanılarak sıçan modelinde araştırıldı. Kırk erkek Sprague-Dawley sıçan beş gruba randomize edildi (n=8). Hepatorenal hasar, beş hafta boyunca haftalık intraperitoneal sisplatin (3 mg/kg) uygulanarak oluşturuldu. 2-APB (2, 4 veya 8 mg/kg, i.p.) eş zamanlı olarak verildi. Serum TNF-α ve LDH düzeyleri, oksidatif stres belirteçleri (MDA, GSH) ve histopatolojik değişiklikler (H&E yarı kantitatif skorlaması; fibrozis için Masson trikrom boyaması) değerlendirildi. Sisplatin uygulaması TNF-α ve LDH düzeylerini anlamlı şekilde artırdı; böbrekte belirgin tübüler nekroz ve karaciğerde hepatoselüler dejenerasyon oluşturdu. 2-APB’nin 2 ve 4 mg/kg dozları biyokimyasal değişiklikleri anlamlı düzeyde azalttı ve histopatolojik hasar skorlarını iyileştirdi. Buna karşın 8 mg/kg dozunda koruyucu etki sınırlı kaldı. Sisplatin GSH düzeylerini artırırken, 2-APB bu kompansatuvar artışı azaltarak oksidatif stresle ilişkili adaptif yanıtın hafifletildiğini düşündürdü. Sonuç olarak, 2-APB’nin 2–4 mg/kg dozları sisplatin kaynaklı hepatorenal toksisiteye karşı etkilidir; 8 mg/kg’da etkinliğin azalması bifazik doz-yanıt ilişkisine işaret etmektedir. Bulgular, 2-APB’nin doz bağımlı bir adjuvan terapötik aday olabileceğini göstermektedir.

Anahtar Kelimeler

• 2-Aminoetoksidifenil borat
• sisplatin
• hepatotoksisite
• nefrotoksisite
• oksidatif stres.

Protective Effects of 2-Aminoethoxydiphenyl Borate on Cisplatin-Induced Hepatorenal Toxicity in Rats

Öz

Cisplatin is a platinum-based chemotherapeutic widely used for solid tumors; however, its clinical utility is limited by dose-dependent hepatotoxicity and nephrotoxicity driven by oxidative stress and disrupted intracellular Ca²⁺ homeostasis. 2-Aminoethoxydiphenyl borate (2-APB), a boron-containing IP₃ receptor antagonist, may exert cytoprotective effects via modulation of calcium signaling. This study investigated the prophylactic potential of 2-APB against cisplatin-induced hepatorenal toxicity in rats using biochemical and histopathological endpoints. Forty male Sprague-Dawley rats were randomized into five groups (n=8). Hepatorenal injury was induced by weekly intraperitoneal cisplatin (3 mg/kg) for five weeks. 2-APB (2, 4, or 8 mg/kg, i.p.) was administered concomitantly. Serum TNF-α and LDH levels, oxidative stress markers (MDA, GSH), and histopathological alterations (H&E scoring; Masson’s trichrome for fibrosis) were evaluated. Cisplatin significantly increased TNF-α and LDH levels and caused marked tubular necrosis in the kidney and hepatocellular degeneration. Treatment with 2-APB at 2 and 4 mg/kg significantly attenuated biochemical alterations and improved histopathological injury scores. In contrast, 8 mg/kg provided limited protection. Although cisplatin elevated GSH levels, 2-APB reduced this compensatory increase, suggesting mitigation of oxidative stress–related adaptive responses. Moderate doses of 2-APB (2–4 mg/kg) effectively protect against cisplatin-induced hepatorenal toxicity, whereas loss of efficacy at 8 mg/kg indicates a biphasic dose-response, potentially due to excessive Ca²⁺ signaling disruption. These findings support 2-APB as a dose-dependent adjunctive therapeutic candidate.

Anahtar Kelimeler

• 2-Aminoethoxydiphenyl borate
• cisplatin
• hepatotoxicity
• nephrotoxicity
• oxidative stress.

Kaynakça

1. 1. Dasari S, Bernard B.T. Cisplatin in cancer therapy: molecular mechanisms of action. Metallics. 2014; 6(11): 1801-1815.
2. 2. Siddik Z.H. Cisplatin: mode of cytotoxic action and molecular basis of resistance. Int J Med. 2003; 21(12): 882-887.
3. 3. Pabla N, Dong Z. Cisplatin nephrotoxicity: mechanisms and renoprotective strategies. Kidney Int. 2008; 73(9): 994-1007.
4. 4. Miller R.P., Tadagavadi R.K., Ramesh G., Reeves W.B. Mechanisms of cisplatin nephrotoxicity. Toxicol Sci. 2010; 118(2): 455-466.
5. 5. Lameire N., Van Biesen W., Vanholder R. Acute renal failure. Lancet. 2008; 372(9642): 181-190.
6. 6. Antunes L.M., Darin J.D., Bianchi N. Effects of the oral administration of zinc on the genotoxicity and levels of cisplatin in the liver and kidney of mice. Toxicol Sci. 2008; 105(2): 367-375.
7. 7. Al-Shorbagy M.Y., Al-Wabel N.A., Al-Quraishi A.A. Cisplatin-induced hepatotoxicity in rats: involvement of oxidative stress and inflammatory mediators. Toxicol Mech Methods. 2015; 25(3): 198-204.
8. 8. Pinar A., Ari N.S., Ozyildirim S. Boron compounds: promising therapeutic agents in different pathological conditions. Mini Rev Med Chem. 2020; 20(13): 1238-1250.

9. 9. Sahin F., Gungor O., Kisa U. The protective effect of boric acid on carbon tetrachloride-induced hepatotoxicity in rats. Biol Trace Elem Res. 2018; 183(1): 127-133.
10. 10. Eker A., Bozkurt A., Yigit I. Sodium pentaborate decahydrate attenuates cisplatin-induced nephrotoxicity in rats. J Trace Elem Med Biol. 2019; 56: 126388.
11. 11. Kilic E., Yigit I., Aksoy A. Protective effects of boron compounds on drug-induced organ toxicities: a systematic review. Chem Biol Interact. 2020; 323: 109062.
12. 12. Maruyama T., Kanaji T., Nakade K., Kanno T., Okada Y. 2-Aminoethoxydiphenyl borate, an $\text{IP}_3$ receptor antagonist, inhibits $\text{Ca}^{2+}$ release at an intracellular site in many cell types. FEBS Lett. 1998; 433(3): 219-222.
13. 13. Bootman M.D., Berridge M.J. $\text{IP}_3$ receptors and the control of calcium signaling. Cell. 2002; 109(2): S155-S166.
14. 14. Ramesh G., Reeves W.B. Cisplatin-induced apoptosis in LLC-PK1 cells: $\text{Ca}^{2+}$ and $\text{Mg}^{2+}$ as modulators. J Am Soc Nephrol. 2002; 13(4): 887-897.
15. 15. Suzuki A., Ohsaka Y., Okada Y. $\text{IP}_3$ receptor-mediated $\text{Ca}^{2+}$ release is involved in cisplatin-induced apoptosis in human proximal tubule cells. Pflugers Arch. 2006; 452(2): 203-211.
16. 16. Li Y., Chen W., Wang H. 2-Aminoethoxydiphenyl borate protects against hydrogen peroxide-induced oxidative stress and apoptosis in H9c2 cardiomyocytes. Redox Rep. 2017; 22(1): 66-72.
17. 17. Choi S.K., Han Y.R., Park J.H. 2-APB inhibits inflammatory responses by suppressing $\text{NF-}\kappa\text{B}$ activation in $\text{LPS}$-stimulated $\text{RAW 264.7}$ macrophages. Int Immunopharmacol. 2019; 72: 185-192.
18. 18. Eser M., Akgul O., Yuce T. Nephroprotective effect of 2-aminoethoxydiphenyl borate against acute kidney injury induced by glycerol in rats. Toxicol Mech Methods. 2021; 31(5): 371-378.Gurel A.,
19. 19. Elbe H., Akbulut B. The effect of 2-aminoethoxydiphenyl borate on experimentally induced liver injury in diabetic rats. Iran J Basic Med Sci. 2020; 23(12): 1612-1618.
20. 20. Buyuk B., Kucuk O., Sen A. Protective role of 2-aminoethoxydiphenyl borate on methotrexate-induced intestinal damage in rats. Dig Dis Sci. 2018; 63(11): 3013-3021.
21. 21. Ahmad N., Subhan F., Islam N.U., et al. Gabapentin and its salicylaldehyde derivative alleviate cisplatin-induced neuropathic pain. Eur J Pharmacol. 2017; 814: 302–312.
22. 22. Koivukoski S., Khan U., Ruusuvuori P., et al. Unstained tissue imaging and virtual H&E staining. Lab Invest. 2023; 103: 100070.
23. 23. Hao Y., Miao J., Liu W., et al. Formononetin protects against cisplatin-induced kidney injury. Int J Mol Med. 2021; 47: 511–522.
24. 24. Arany I., Safirstein R.L. Cisplatin nephrotoxicity. Semin Nephrol. 2003; 23: 460–464.
25. 25. Jang D.I., Lee A.H., Shin H.Y., et al. Role of TNF-$\alpha$ in autoimmune disease and inhibitors. Int J Mol Sci. 2021; 22: 2719.
26. 26. Halliwell B., Gutteridge J.M.C. Free radicals in biology and medicine. 5th ed. Oxford University Press, Oxford; 2015.
27. 27. Anderson M.E. Glutathione: an overview of biosynthesis and modulation. Chem Biol Interact. 1998; 111–112: 1–14.
28. 28. Eroglu E, Unel CC, Harmanci N, Erol K, Ari NS, Ozatik O. 2-Aminoethoxydiphenyl borate ameliorates functional and structural abnormalities in cisplatin-induced peripheral neuropathy. J Trace Elem Med Biol. 2022; 70: 126909.
29. 29. Iqbal N, Umar S, Khan NA, Corpas FJ. Nitric Oxide and Hydrogen Sulfide Coordinately Reduce Glucose Sensitivity and Decrease Oxidative Stress via Ascorbate-Glutathione Cycle in Heat-Stressed Wheat (Triticum aestivum L.) Plants. Antioxidants (Basel). 2021; 10(1): 108.
30. 30. Gugliandolo E, Fusco R, Biundo F, D'Amico R, Benedetto F, Di Paola R, Cuzzocrea S. Palmitoylethanolamide and Polydatin combination reduces inflammation and oxidative stress in vascular injury. Pharmacol Res. 2017; 123: 83-92.
31. 31. Wang M, Fang L, Liu T, Chen X, Zheng Y, Zhang Y, Chen S, Li Z. Discovery of 7-O-1, 2, 3-triazole hesperetin derivatives as multi-target-directed ligands against Alzheimer's disease. Chem Biol Interact. 2021; 342: 109489.
32. 32. Mansour H, Ata M, Mahran A, Hasan N, AboBakr AH. Protective Effect of Melatonin on Cisplatin Induced Liver Toxicity in Albino Rats; Biochemical and Histological Study. Minia Journal of Medical Research. 2024; 35(4): 57-68.
33. 33. Iwasaki H, Mori Y, Hara Y, Uchida K, Zhou H, Mikoshiba K. 2-Aminoethoxydiphenyl borate (2-APB) inhibits capacitative calcium entry independently of the function of inositol 1,4,5-trisphosphate receptors. Recept Channels. 2001; 7(6): 429-39.
34. 34. Pabla N, Dong Z. Cisplatin nephrotoxicity: mechanisms and renoprotective strategies. Kidney Int. 2008; 73(9): 994-1007.
35. 35. Slowik EJ, Stankoska K, Bui NN, Pasieka B, Conrad D, Zapp J, Hoth M, Bogeski I, Kappl R. The calcium channel modulator 2-APB hydrolyzes in physiological buffers and acts as an effective radical scavenger and inhibitor of the NADPH oxidase 2. Redox Biol. 2023; 61: 102654.
36. 36. Oun R, Wallace S, Medline A. Cisplatin-induced nephrotoxicity: a review on current strategies and future directions. Journal of Renal Failure. 2016.
37. 37. Schuchter LM, Luginbuhl TG, Meropol NJ. Amifostine: a cytoprotective agent with multiple mechanisms of action. Clin Cancer Res. 2005; 11(5): 1692-700.