Sıçanlarda doksorubisin kaynaklı testis hasarına karşı crocinin faydalı rolü: vimentin modülasyonuna ilişkin görüşler

Year 2024, Volume: 49 Issue: 2, 332 - 342, 30.06.2024

Melike Özgül Önal , Sara Asaad , Gürkan Yiğittürk , Volkan Yaşar , Yasemin Biçer , Dilan Çetinavcı , Eyüp Altınöz , Mehmet Demir , Hülya Elbe , Feral Öztürk

https://doi.org/10.17826/cumj.1420141

Abstract

Amaç: Doksorubisin (DOX) kemoterapide kullanılan geniş spektrumlu bir antibiyotiktir. Yan etkileri tedaviyi kısıtlamaktadır. Crocin, hem antiinflamatuar hem de antioksidan aktiviteye sahip karotenoidlerden biridir. Sıçanlarda doksorubisin kaynaklı testis hasarına karşı Crosinin etkilerini değerlendirmeyi amaçladık.
Gereç ve Yöntem: Kırk adet Wistar sıçan dört gruba ayrıldı. Grup 1: Kontrol, Grup 2: Crosin, Grup 3: DOX, Grup 4: DOX+Crosin (tümü için, n=10). Testis dokuları Hematoksilen-Eozin ile boyandı. Seminifer tübüllerin çapları ölçüldü ve testisin ortalama histopatolojik hasar skoru (MHDS) ölçüldü. Sertoli hücrelerinde Vimentin ekspresyonu H-Score olarak hesaplandı. Testis dokularında Malondialdehit (MDA) ve Glutatyon (GSH), Katalaz (CAT) ve Süperoksit dismutaz (SOD) aktivitelerinin düzeyleri belirlendi. Toplam Antioksidan Durumu (TAS) ve Toplam Oksidan Durumu (TOS) da hesaplandı.
Bulgular: DOX grubunda atrofik seminifer tübüller görüldü. Hasarlanan tübüllerde ödem, vakuolizasyon ve dezorganizasyon mevcuttu. DOX grubu ve kontrol grupları için MHDS'ler sırasıyla 4,60±0,45 ve 0,20±0,13 idi. Bu grupların her ikisi de anlamlı bir fark gösterdi. Crocin kullanımı sonrası histopatolojik hasar skoru azaldı. DOX grubuyla karşılaştırıldığında DOX+Crosin grubunda Vimentin immün ekspresyon düzeyleri ve seminifer tübül genişliği anlamlı derecede artarken tübül hasarı önemli ölçüde azaldı. DOX tedavisi sonrasında MDA ve TOS düzeyleri anlamlı düzeyde artarken, GSH, SOD, CAT ve TAS düzeyleri anlamlı düzeyde azaldı. Crocin aldıktan sonra tüm biyokimyasal göstergeler büyük ölçüde iyileşti.
Sonuç: Crosin takviyesi, oksidan/antioksidan durumunu dengeleyerek DOX'un neden olduğu fonksiyonun testis hasarına karşı yararlı etkiler gösterdi.

Keywords

Doksorubisin, Crosin, testis, vimentin, oksidatif stres.

Supporting Institution

Karabuk University Scientifict Research Fund

Project Number

TYL-2020-2298

Beneficial role of crocin against doxorubicin-induced testicular damage in rats: insights into vimentin modulation

Abstract

Purpose: Doxorubicin (DOX) is a wide-spectrum antibiotic used for chemotherapy. Its side effects limit treatment. Crocin is one of the carotenoids that has both anti-inflammatory and antioxidant activities. We aimed to evaluate the effects of crocin against doxorubicin-induced testicular damage in rats.
Materials and Methods: Forty Wistar rats were divided into four groups. Group 1: Control, Group 2: Crocin, Group 3: DOX, Group 4: DOX+Crocin (n=10, for all). Testis tissues were stained with Hematoxylin-Eosin. The diameters of seminiferous tubules were measured and the testicular mean histopathologic damage score (MHDS) was calculated. Vimentin expression in Sertoli cells was calculated as H-Score. Levels of Malondialdehyde (MDA), Glutathione (GSH), Catalase (CAT), and Superoxide dismutase (SOD) activities were determined in testis tissues. Total Antioxidant Status (TAS) and Total Oxidant Status (TOS) were also calculated.
Results: Atrophic seminiferous tubules were seen in the DOX group. Edema, vacuolization, and disorganization were present in the injured tubules. The MHDSs for the DOX group and control groups were 4.60±0.45 and 0.20±0.13, respectively. Both of these groups showed a significant difference. The histopathologic score was reduced after using crocin. Tubule damage considerably decreased while immunoexpression levels of vimentin and seminiferous tubule width significantly increased in the DOX+Crocin group compared to the DOX group. MDA and TOS levels were significantly increased after DOX treatment, and GSH, SOD, CAT, and TAS levels were significantly decreased. All biochemical indicators were greatly improved after receiving crocin.
Conclusion: Crocin supplementation exhibited adequate beneficial effects against the testicular damage of DOX-induced function by balancing the oxidant/antioxidant status.

Keywords

Doxorubicin, crocin, testis, vimentin, oxidative stress.

Ethical Statement

The Zonguldak Bulent Ecevit University Experimental Animals Ethics Committee approved the study (Protocol No. 2021-09-01/04).

Supporting Institution

The research was supported by Karabuk University Scientifict Research Fund.

Project Number

TYL-2020-2298

References

• Arcamone F, Cassinelli G, Fantini G, Grein A, Orezzi P, Pol C et al. Adriamycin, 14‐Hydroxydaunomycin, a new antitumor antibiotic from S. peucetius var. caesius. Biotechnol Bioeng. 2000;67:704-13.
• Shinoda K, Mitsumori K, Yasuhara K, Uneyama C, Onodera H, Hirose M et al. Doxorubicin induces male germ cell apoptosis in rats. Arch Toxicol. 1999;73:274-81.
• Hou MI, Chrysis D, Nurmio M, Parvinen M, Eksborg S, Söder O et al. Doxorubicin induces apoptosis in germ line stem cells in the immature rat testis and amifostine cannot protect against this cytotoxicity. Cancer Res. 2005;65:9999-10005.
• Rivankar S. An overview of doxorubicin formulations in cancer therapy. J Cancer Res Ther. 2014;10:853.
• Lalmuansangi C, Zosangzuali M, Lalremruati M, Tochhawng L, Siama Z. Evaluation of the protective effects of Ganoderma applanatum against doxorubicin-induced toxicity in Dalton's lymphoma ascites (DLA) bearing mice. Drug Chem Toxicol. 2022;45:1243-53.
• Yeh YC, Liu TJ, Wang LC, Lee HW, Ting CT, Lee WL et al. A standardized extract of Ginkgo biloba suppresses doxorubicin‐induced oxidative stress and p53‐mediated mitochondrial apoptosis in rat testes. Br J Pharmacol. 2009;156:48-61.
• Yang CC, Chen YT, Chen CH, Chiang JY, Zhen YY, Yip HK. Assessment of doxorubicin-induced mouse testicular damage by the novel second-harmonic generation microscopy. Am J Transl Res. 2017;9:5275-88.
• Rizk SM, Zaki HF, Mina MAM. Propolis attenuates doxorubicin-induced testicular toxicity in rats. Food Chem Toxicol. 2014;67:176-86.
• Mzabri I, Addi M, Berrichi A. Traditional and modern uses of saffron (Crocus sativus). Cosmetics. 2019;6:63.
• Lee IA, Lee JH, Baek NI, Kim DH. Antihyperlipidemic effect of crocin isolated from the fructus of Gardenia jasminoides and its metabolite crocetin. Biol Pharm Bull. 2005;28:2106-10.
• Zheng YQ, Liu JX, Wang JN, Xu L. Effects of crocin on reperfusion-induced oxidative/nitrative injury to cerebral microvessels after global cerebral ischemia. Brain Research. 2007;1138:86-94.
• Hassani FV, Naseri V, Razavi BM, Mehri S, Abnous K, Hosseinzadeh H. Antidepressant effects of crocin and its effects on transcript and protein levels of CREB, BDNF, and VGF in rat hippocampus. DARU. 2014; 22:1-9.
• Wang G, Zhang B, Wang Y, Han S, Wang C. Crocin promotes apoptosis of human skin cancer cells by inhibiting the JAK/STAT pathway. Exp Ther Med. 2018;16:5079-84.
• Wang C, Cai X, Hu W, Li Z, Kong F, Chen X et al. Investigation of the neuroprotective effects of crocin via antioxidant activities in HT22 cells and in mice with Alzheimer's disease. Int J Mol Med. 2019;43:956-66.
• Premkumar K, Abraham SK, Santhiya ST, Gopinath PM, Ramesh A. Inhibition of genotoxicity by saffron (Crocus Sativus L.) in mice. Drug Chem Toxicol. 2001;24:421-8.
• Boussada M, Dias TR, Crisóstomo L, Akacha AB, Ali RB, Michèle V et al. A new thiocyanoacetamide (2-cyano-2-p-nitrophenyl-N-benzylthioamide) reduces doxorubicin-induced in vitro toxicity in Sertoli cells by decreasing apoptosis and autophagy. Theriogenology. 2019;140:188-200.
• Huyut Z, Alp HH, Yaman T, Keleş ÖF, Yener Z, Türkan F, Ayengin K. Comparison of the protective effects of curcumin and caffeic acid phenethyl ester against doxorubicin-induced testicular toxicity. Andrologia. 2021;53:13919.
• Kaur S, Maan KS, Sadwal S, Aniqa A. Studies on the ameliorative potential of dietary supplemented selenium on doxorubicin-induced testicular damage in mice. Andrologia. 2020;52:13855.
• Devkota B, Sasaki M, Matsui M, Takahashi KI, Matsuzaki S, Koseki T et al. Effects of scrotal insulation and pathological lesions on α-smooth muscle actin (SMA) and vimentin in the bull testes. J Reprod Dev. 2010; 56:187-90.
• ElGhamrawy TA, Helmy D, Elall HF. Cadherin and vimentin immunoexpression in the testis of normal and induced infertility models of albino rats. Folia Morphol (Warsz). 2014;73:339-46.
• Razmaraii N, Babaei H, Nayebi AM, Asadnasab G, Helan JA, Azarmi Y. Cardioprotective effect of phenytoin on doxorubicin-induced cardiac toxicity in a rat model. Journal Of Cardiovasc Pharmacol. 2016;67:237-45.
• Razmaraii N, Babaei H, Nayebi AM, Assadnassab G, Helan JA, Azarmi Y. Crocin treatment prevents doxorubicin-induced cardiotoxicity in rats. Life Sci. 2016;157:145-51.
• Hirsch FR, Varella-Garcia M, Bunn Jr PA, Di Maria MV, Veve R, Bremnes RM et al. Epidermal growth factor receptor in non–small-cell lung carcinomas: correlation between gene copy number and protein expression and impact on prognosis. J Clin Oncol. 2003;21:3798-807.
• Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95:351-8.
• Sun YI, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem. 1988;34:497-500.
• Aebi H. Catalase. In Methods of Enzymatic Analysis (Ed. HU Bergmeyer):673-84. New York, Academic Press, 1974.
• Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959;82:70-7.
• Gornall AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem. 1949;177:751-66.
• Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem. 2004; 37:277-85.
• Erel O. A new automated colorimetric method for measuring total oxidant status. Clin Biochem. 2005;38:1103-11.
• Levi M, Tzabari M, Savion N, Stemmer SM, Shalgi R, Ben-Aharon I. Dexrazoxane exacerbates doxorubicin-induced testicular toxicity. Reproduction (Cambridge, England). 2015; 150:357-66.
• Mohan UP, PB TP, Iqbal ST, Arunachalam S. Mechanisms of doxorubicin-mediated reproductive toxicity–a review. Reprod Toxicol. 2021;102:80-9.
• Brilhante O, Okada FK, Sasso-Cerri E, Stumpp T, Miraglia SM. Late morfofunctional alterations of the Sertoli cell caused by doxorubicin administered to prepubertal rats. Reprod Biol Endocrinol. 2012;10:1-6.
• Tremblay AR, Delbes G. In vitro study of doxorubicin-induced oxidative stress in spermatogonia and immature Sertoli cells. Toxicol Appl Pharmacol. 2018;348:32-42.
• Belhan S, Özkaraca M, Özdek U, Kömüroğlu AU. Protective role of chrysin on doxorubicin-induced oxidative stress and DNA damage in rat testes. Andrologia. 2020;52:13747.
• Martin RH, Rademaker AW, Leonard NJ. Analysis of chromosomal abnormalities in human sperm after chemotherapy by karyotyping and fluorescence in situ hybridization (FISH). Cancer Genet Cytogenet. 1995;80:29-32.
• Öztürk E, Kaymak EM, Akin AT, Karabulut DE, Ünsal HM, Yakan Bİ. Thymoquinone is a protective agent that reduces the negative effects of doxorubicin in rat testis. Hum Exp Toxicol. 2020;39:1364-73.
• Lee KM, Lee IC, Kim SH, Moon C, Park SH, Shin DH et al. Melatonin attenuates doxorubicin‐induced testicular toxicity in rats. Andrologia. 2012;44:796-803.
• Kopecky M, Semecky V, Nachtigal P. Vimentin expression during altered spermatogenesis in rats. Acta Histochemic. 2005;107:279-89.
• Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis. 2005;15:316-28.
• Kayar A, Dokuzeylul B, Kandemir FM, Kirbas A, Bayrakal A, Or ME. Total oxidant and antioxidant capacities, nitric oxide and malondialdehyde levels in cats seropositive for the feline coronavirus. Veterinarni Medicina. 2015;60:274-81.
• McLennan SV, Heffernan S, Wright L, Rae C, Fisher E, Yue DK et al. Changes in hepatic glutathione metabolism in diabetes. Diabetes.1991;40:344-8.
• El-Maddawy ZK, Abd El Naby WS. Protective effects of zinc oxide nanoparticles against doxorubicin induced testicular toxicity and DNA damage in male rats. Toxicol Res (Camb). 2019;8:654-62.
• Mohamed RH, Karam RA, Hagrass HA, Amer MG, Abd El-Haleem MR. Anti-apoptotic effect of spermatogonial stem cells on doxorubicin-induced testicular toxicity in rats. Gene. 2015;561:107-14.
• Farsani BE, Karimi S, Mansouri E. Pravastatin attenuates testicular damage induced by doxorubicin–a stereological and histopatological study. J Basic Clin Physiol Pharmacol. 2019; 30:103-9.
• Chu X, Zhang Y, Xue Y, Li Z, Shi J, Wang H et al. Crocin protects against cardiotoxicity induced by doxorubicin through TLR-2/NF-κB signal pathway in vivo and vitro. Int Immunopharmacol. 2020;84:106548.
• Gül M, Kayhan Kuştepe E, Erdemli ME, Altınöz E, Gözükara Bağ HG, Gül S et al. Protective effects of crocin on acrylamide‐induced testis damage. Andrologia. 2021;53:14176.
• Mesbahzadeh B, Hassanzadeh-Taheri M, Aliparast MS, Baniasadi P, Hosseini M. The protective effect of crocin on cisplatin-induced testicular impairment in rats. BMC Urol. 2021;21:1-9.
• Elsherbiny NM, Salama MF, Said E, El-Sherbiny M, Al-Gayyar MM. Crocin protects against doxorubicin-induced myocardial toxicity in rats through down-regulation of inflammatory and apoptic pathways. Chem Biol Interact. 2016;247:39-48.
• Abdulkareem Aljumaily SA, Demir M, Elbe H, Yigitturk G, Bicer Y, Altinoz E. Antioxidant, anti-inflammatory, and anti-apoptotic effects of crocin against doxorubicin-induced myocardial toxicity in rats. Environ Sci Pollut Res Int. 2021;28:65802-13.
• Vafaei S, Motejaded F, Ebrahimzadeh-Bideskan A. Protective effect of crocin on electromagnetic field-induced testicular damage and heat shock protein A2 expression in male BALB/c mice. Iran J Basic Med Sci. 2020;23:102.
• Sefidgar SM, Ahmadi-Hamedani M, Javan AJ, Sani RN, Vayghan AJ. Effect of crocin on biochemical parameters, oxidative/antioxidative profiles, sperm characteristics and testicular histopathology in streptozotocin-induced diabetic rats. Avicenna J Phytomed. 2019;9:347.
• Sadat Kamali F, Shahrooz R, Najafi G, Razi M. Ameliorative effects of crocin on paraquat-induced oxidative stress in testis of adult mice: An experimental study. Int J Reprod Biomed. 2019;17:807-18.
• Davoodi M, Bouri SZ, Ghahfarokhi SD. Antioxidant effects of aerobic training and crocin consumption on doxorubicin-induced testicular toxicity in rats. J Family Reprod Health . 2021;5:28-37.