Protective effects of boric acid possessing antioxidant and cytoprotective properties against cyclophosphamide-induced liver damage in rats: A histopathological and stereological study

Year 2025, Volume: 52 Issue: 3, 453 - 462, 16.09.2025

Elfide Gizem Bakirhan Ebru Annaç , Elif Merve Betül Yanılmaz , Nurhan Tirasci

https://doi.org/10.5798/dicletip.1784951

Abstract

Objective: Our study was conducted to investigate the potential protective activities of boric acid (BA), which exhibits antioxidant and protective properties against cyclophosphamide (CP)-induced liver injury in rats, using histopathological and stereological methods.
Methods: Twelve-week-old rats were randomly assigned to one of four groups of eight animals each: Control (Cont: receiving 1 ml tap water by oral gavage for 7 days), CP (a single intraperitoneal (i.p.) dose of 150 mg/kg, BA (15 mg/kg/day, orally for 7 days), and CP+BA (a single dose of 150 mg/kg CP i.p., plus 15 mg/kg/day BA by oral gavage for 7 days). At the end of the experiment, liver tissues were collected after anesthesia and decapitation. Total liver and sinusoidal volumes were estimated using the Cavalieri method. Hematoxylin-eosin, Masson trichrome, and periodic acid-Schiff (PAS) staining were applied for histopathological evaluations.
Results: Stereological analysis revealed a statistically significant increase in total liver volume in the CP group compared to the Cont (p=0.004) and CP+BA groups (p=0.045). Sinusoidal volume was also significantly higher in the CP group compared to the Cont (p=0.001) and CP+BA groups (p=0.037). Histopathological examination revealed impaired hepatic architecture, hepatocyte degeneration, and increased sinusoidal dilatation and mast cell density in the CP group. In contrast, a partially preserved liver histology with reduced degeneration was observed in the CP+BA group.
Conclusion: BA treatment exhibits a partial hepatoprotective effect against CP-induced liver injury, as shown by both the stereological and histopathological analyses. The study findings indicate the hepatoprotective potential of BA treatment and suggest that it represents a promising candidate for therapeutic use.

Keywords

Boric acid , Cyclophosphamide , Liver damage

Ethical Statement

Ethical protocol was approved by Adıyaman University Ethics Committee, approval number (Decision No: 2024/017-08).

Sıçanlarda siklofosfamid kaynaklı karaciğer hasarına karşı antioksidan ve sitoprotektif özelliklere sahip borik asidin koruyucu etkileri: Histopatolojik ve stereolojik bir çalışma

Abstract

Amaç: Çalışmamızda, histopatolojik ve stereolojik yöntemler kullanarak sıçanlarda siklofosfamid (CP) kaynaklı karaciğer hasarına karşı antioksidan ve koruyucu özelliklere sahip olan borik asidin (BA) potansiyel koruyucu rolünü değerlendirmeyi amaçlamaktadır.
Yöntemler: Çalışmamızda 12 haftalık sıçanı rastgele dört gruba ayrıldı (n=8): Kontrol (Cont: 7 gün boyunca oral gavaj ile 1ml çeşme suyu), CP (tek doz 150 mg/kg, i.p.), BA (15 mg/kg/gün, 7 gün boyunca oral) ve CP+BA (tek doz 150 mg/kg, i.p CP, 7 gün boyunca oral gavaj ile 15 mg/kg/gün BA). Deney sonuunda, anestezi ve dekapitasyonun ardından karaciğerdokuları toplandı. Toplam karaciğer ve sinüzoidal hacimler Cavalieri yöntemi kullanılarak tahmin edildi. Histopatholojikdeğerlendirmeler için hematoksilen-Eozin, Masson trikromu ve PAS boyamalar yapıldı.
Bulgular: Stereolojik analiz, CP grubunda Cont (p=0.004) ve CP+BA (p=0.045) gruplarına kıyasla toplam karaciğer hacminde istatistiksel olarak anlamlı bir artış olduğu gözlendi. Sinüzoidal hacim de CP grubunda Cont (p=0.001) ve CP+BA (p=0.037) gruplarına kıyasla anlamlı bir şekilde arttış görüldü. Histopatolojik olarak, CP grubunda bozulmuş hepatik mimari, hepatosit dejenerasyonu ve artmış sinüzoidal dilatasyon ve mast hücre yoğunluğu görüldü. Buna karşılık, CP+BA grubunda dejenerasyonun azaldığı kısmen korunmuş karaciğer histolojisi görüldü.
Sonuç: BA tedavisi, hem stereolojik hem de histopatolojik analizlerle gösterildiği üzere, CP kaynaklı karaciğer hasarına karşı kısmi hepatoprotektif etki göstermektedir. Çalışmamız, BA tedavisi hepatoprotektif potansiyelini ortaya koymakta ve olası terapötik kullanımı açısından umut verici bir aday olduğunu düşündürmektedir.

Keywords

Borik ait , Siklofosfamid , Karaciğer hasarı

References

• 1.Asiri YA. Probucol attenuates cyclophosphamide-induced oxidative apoptosis, p53 and Baxsignalexpression in rat cardiac tissues. Oxid Med CellLongev. 2010; 3: 308–16.
• 2. Emadi A, Jones RJ, Brodsky RA. Cyclophosphamide and cancer: golden anniversary. Nat Rev Clin Oncol.2009; 6: 638–47.
• 3.Şahin F, Deveci E, Aşır F, Albayrak MGB,Özkorkmaz EG. Investigation of the effect ofrosmarinic acid on cyclophosphamide-inducedgonadal toxicity. ASM. 2022; 12: 1-8.
• 4.Madondo MT, Quinn M, Plebanski M. Low dosecyclophosphamide: Mechanisms of T cellmodulation. Cancer Treat Rev. 2016; 42: 3–9.
• 5.Abdelfattah-Hassan A, Shalaby SI, Khater SI, et. al.Panax ginseng is superior to vitamin E as ahepatoprotector against cyclophosphamide-induced liver damage. Complement Ther Med. 2019;46: 95–102.
• 6.Nuño-Lámbarri N, Domínguez-Pérez M, Baulies-Domenech A, et al. liver cholesterol overloadaggravates obstructive cholestasis by inducingoxidative stress and premature death in mice. OxidMed Cell Longev. 2016; 2016: 9895176.
• 7.Hao H, Xu Y, Chen R, et al. Protective effects ofchlorogenic acid against cyclophosphamide inducedliver injury in mice. Biotec Histochem. 2024; 99: 33–43.
• 8.Habibi E, Shokrzadeh M, Chabra A, et al. Protective effects of Origanum vulgare ethanol extract againstcyclophosphamide-induced liver toxicity in mice.Pharm Biol. 2015; 53: 10–5.
• 9.Cengiz M. Boric acid protects againstcyclophosphamide-induced oxidative stress andrenal damage in rats. Cell Mol Biol. 2018; 64: 11–4.
• 10.Hadrup N, Frederiksen M, Sharma AK. Toxicity ofboric acid, borax and other boron containingcompounds: A review. RTP. 2021; 121:104873.
• 11. Basbug M, Yildar M, Yaman İ, et al. Effects of boricacid in an experimental rat model of hepaticischemia-reperfusion injury. Acta Med. 2015;31:1067-1073.
• 12.Abogresha NM, Mohammed SS, Hosny MM, et al.Diosmin mitigates cyclophosphamide inducedpremature ovarian insufficiency in rat model. Int JMol Sci. 2021; 22: 1–20.
• 13.Çolak S, Koc K, Yıldırım S, Geyikoğlu F. Effects ofboric acid on ovarian tissue damage caused byexperimental ischemia/reperfusion. BiotecHistochem. 2022; 97: 415–22.
• 14.Bakirhan EG, Yanilmaz EMB, Tüfekci KK,Bakirhan F, Susam S. Maternal high-fat diet impairscognitive performance by altering hippocampalGRP78/PERK axis and BDNF expression in adultfemale rat offspring: the potential protective role ofN acetylcysteine. J Mol Histol. 2025; 56: 189.
• 15.Alqahtani S, Mahmoud AM. Gamma-glutamylcysteine ethyl ester protects againstcyclophosphamide-induced liver ınjury andhematologic alterations via upregulation of PPARγand attenuation of oxidative stress, ınflammation,and apoptosis. Oxid Med Cell Longev. 2016; 2016:4016209.
• 16.Kızılay Z, Erken HA, Çetin NK, et al. Boric acidreduces axonal and myelin damage in experimentalsciatic nerve injury. Neural Regen Res. 2016;11:1660–5.
• 17.Coban FK, Ince S, Kucukkurt I, Demirel HH,Hazman O. Boron attenuates malathion-inducedoxidative stress and acetylcholinesterase inhibitionin rats. Drug Chem Toxicol. 2015; 38: 391–9.
• 18.Rezaei S, Hosseinimehr SJ, Zargari M, et al.Sinapic acid attenuates cyclophosphamide-inducedliver toxicity in mice by modulating oxidative stress,NF-κB, and caspase-3. Iran J Basic Med Sci. 2023; 26:526.
• 19.Li X, Li B, Jia Y. The hepatoprotective effect ofhaoqin qingdan decoction against liver injuryinduced by a chemotherapeutic drug cyclophosphamide. eCAM. 2015; 2015: 978219.
• 20.Caglayan C, Temel Y, Kandemir FM, Yildirim S,Kucukler S. Naringin protects againstcyclophosphamide-induced hepatotoxicity andnephrotoxicity through modulation of oxidativestress, inflammation, apoptosis, autophagy, andDNA damage. ESPR. 2018; 25: 20968–84.
• 21.Güler S, Aslaner A, Ellidağ HY, Yildirim Ş, Çakir T.The protective effect of boric acid on cholestatic ratliver ischemia reperfusion injury. Turk J Med Sci.2021; 51: 2716. https://doi.org/10.3906/SAG-2101-153.
• 22.Kahraman E, Göker E. Boric acid exerts anti-cancer effect in poorly differentiated hepatocellularcarcinoma cells via inhibition of AKT signalingpathway. J Trace Elem Medi Biol. 2022; 73: 127043.
• 23.Cengiz M, Cetik Yildiz S, Demir C, et al. Hepato-preventive and anti-apoptotic role of boric acidagainst liver injury induced by cyclophosphamide. JTrace Elem Med Biol. 2019; 53: 1–7.
• 24.Önder Gö, Göktepe Ö, Okur E, et al. Boric acidameliorates liver injury in rat induced bycyclophosphamide. Sakarya Med J. 2023; 13: 210-6.
• 25.Gündüz E, Yildizhan E, Yaman M, Sen A, Akkus M.Investigation of the protective effect of boric acidagainst hepatotoxic and nephrotoxic injury inducedby acrylamide in rats. Int J Morphol. 2023; 41: 368-73.