4-Experimental Surgery The therapeutic effects of curcumin and capsaicin against cyclophosphamide side effects on the uterus in rats1 Ercan YilmazI, Rauf MelekogluII, Osman CiftciIII, Sevil EraslanIV, Asli CetinV, Nese BasakVI IAssociate Professor, Medicine Faculty, Inonu University, Department of Obstetrics and Gynecology, Malatya, Turkey. Manuscript writing. IIAssistant Professor, Medicine Faculty, Inonu University, Department of Obstetrics and Gynecology, Malatya, Turkey. Acquisition of data. IIIFull Professor, Medicine Faculty, Pamukkale University, Department of Medical Pharmacology, Denizli, Turkey. Analysis of data. IVMD, Elbistan State Hospital, Department of Obstetrics and Gynecology, Kahramanmaras, Turkey. Statistical analysis. VAssistant Professor, Medicine Faculty, Inonu University, Department of Histology, Malatya, Turkey. Histopathological analysis. VIMD, Pharmacy Faculty, Inonu University, Department of Pharmeceutical Toxicology, Malatya, Turkey. Acquisition of data. Abstract Purpose: To evaluate the impact of systemic cyclophosphamide treatment on the rat uterus and investigate the potential therapeutic effects of natural antioxidant preparations curcumin and capsaicin against cyclophosphamide side effects. Methods: A 40 healthy adult female Wistar albino rats were used in this study. Rats were randomly divided into four groups to determine the effects of curcumin and capsaicin against Cyclophosphamide side effects on the uterus (n=10 in each group); Group 1 was the control group (sham-operated), Group 2 was the cyclophosphamide group, Group 3 was the cyclophosphamide + curcumin (100mg/kg) group, and Group 4 was the cyclophosphamide + capsaicin (0.5 mg/kg) group. Results: Increased tissue oxidative stress and histological damage in the rat uterus were demonstrated due to the treatment of systemic cyclophosphamide chemotherapy alone. The level of tissue oxidant and antioxidant markers and histopathological changes were improved by the treatment of curcumin and capsaicin. Conclusion: Cytotoxic effects of natural alkylating chemotherapeutic agents like cyclophosphamide on the uterus can be prevented by curcumin and capsaicin. Key words: Cyclophosphamide. Curcumin. Capsaicin. Antioxidant. Rats. DOI: http://dx.doi.org/10.1590/s0102-865020180060000004 Acta Cir Bras. 2018;33(6):499-507 499 The therapeutic effects of curcumin and capsaicin against cyclophosphamide Acta Cir Bras. 2018;33(6):499-507 side effects on the uterus in rats Yilmaz E et al. ■ Introduction topical medicines all over the world and has anti-inflammatory, antioxidant and anti- It is now accepted that the incidence of tumoral properties5. cancers of the organs and tissues has increased Although the antitumoral properties of significantly in recent years throughout both compounds are known, how they prevent the world1. As is known, adjuvant and/or complex tumorigenic pathways characterized neoadjuvant chemotherapeutic agents have by uncontrolled cell proliferation, angiogenesis, an essential role in addition to appropriate invasion, and metastasis is still unclear. It surgical treatment in cancer therapy. Besides is believed that this effect is achieved by the ability of chemotherapeutic agents inhibiting the transcription and growth factors, to neutralize tumor cells, they also have blocking the receptor, inactivating the kinase significant adverse effects on many cells in the enzyme system, and inhibiting cytokine organism, especially those with anti-mitotic release which causes tumorigenesis. Similarly, potency2. the catastrophic effect of chemotherapeutic Cyclophosphamide is an alkylating agents on the body is thought to be inhibited chemotherapeutic agent, and it has an by the mechanisms outlined above6. extensive use for adjuvant or neoadjuvant In this study, we aimed to show the purposes in the treatment of hematological beneficial effect of CRC and CPS treatment malignancies, neuroblastoma, retinoblastoma, against the tissue damage due to systemic rhabdomyosarcoma, breast, lung, cyclophosphamide chemotherapy in rat uterus. endometrium and ovarian cancers. Despite its frequent use, catastrophic side effects ■ Methods on the genitourinary, reproductive and gastrointestinal system organs are still not The experiments were performed preventable today3. While the effect of CYC on according to the animal ethics guidelines ovary in the female genital system has been of the Inonu University Institutional Animal well described, there is limited data about the Ethics Committee (Ethical approval no: effect of CYC on the uterus. 2016/A-87). The use of natural antioxidants A total of 40 healthy adult female Wistar in order to minimize the side effects of albino rats (aged 3–4 months) obtained from chemotherapeutic agents such as CYC has The Experimental Animal Institute (Malatya, increased in recent years. Curcumin (CRC) and Turkey) were used in the experiment. Animals capsaicin (CPS) are the most prominent of were housed in sterilized polypropylene rat these. CRC is derived from “Curcuma longa”. cages under a 12/12-h light/dark cycle at an Its homeland is East India and is defined as ambient temperature of 21°C. Food and water “the queen of spices”. It includes natural were provided ad libitum. Rats were randomly polyphenol derivatives, gives a yellow color divided into four groups to determine the to natural turmeric plant, has anti-oxidant, effects of curcumin and capsaicin against anti-inflammatory, and apoptosis-inducing cyclophosphamide side effects on the uterus features and naturally inhibits cancer cell (n=10 in each group); Group 1 was the proliferation4. CPS is a trans-8-methyl-N- control group (sham-operated), Group 2 was vanillyl-6-nonenamide compound obtained the cyclophosphamide group, Group 3 was from chili pepper that is found in spices and the cyclophosphamide + curcumin (100mg/ 500 Acta Cir Bras. 2018;33(6):499-507 The therapeutic effects of curcumin and capsaicin against cyclophosphamide side effects on the uterus in rats Yilmaz E et al. kg) (CYC+CRC) group, and Group 4 was the thick sections, mounted on slides, and stained cyclophosphamide + capsaicin (0.5 mg/kg) with hematoxylin and eosin (H-E). The tissue (CYC+CPS) group. Chemotherapy induction samples were examined using a Leica DFC280 was carried out by intraperitoneal injection light microscope and a Leica Q-Win Image of 200 mg/kg CYC (Endoxan, EIP ECZACIBASI, Analysis system (Leica Micros Imaging Solutions Istanbul, Turkey) on day 1 and then 8 mg/ Ltd., Cambridge, UK). Histopathological kg/day for the following 14 days to the rats examination of the tissue damage was in group 2, 3, and 4. After 14 days curcumin performed regarding each parameter, (100mg/kg) and capsaicin (0.5 mg/kg) were such as inflammatory cells in the lumen, orally administered for 14 days by gavages. The vascular congestion, epithelial degeneration doses of CRC and CPS were chosen based on and vacuoles in epithelial cells and dilated previous studies7,8. No treatment was given to endometrial gland. At least five microscopic the control group. The animals were sacrificed regions were examined to score the specimens without pain or distress using increasing semi-quantitatively. Each sample was scored anaesthesia doses (intraperitoneal lethal doses for each criterion using a scale ranging from 0 of pentobarbital; Bioveta Inc., Ankara, Turkey) to 3 (0, none; 1, mild; 2, moderate; 3, severe). after the treatment course, and the uterine Total scores were calculated according to these horns were removed for histopathological parameters. analyses. Blood samples were collected under anaesthesia from the left ventricle using a Statistical analysis syringe. Tissue samples were stored at −45°C All values were presented as the until analysed. mean ± standard deviation. Differences Biochemical assays were considered significant at p < 0.01. The SPSS software (ver. 18.0; SPSS Inc., Chicago, Tissues were homogenized using IL, USA) was used for statistical analyses. a Teflon glass homogenizer in 150 mM Biochemical values were analyzed using one- KCl (pH 7.4) at a 1:10 (w/v) dilution of the way ANOVA and post hoc Tukey’s honestly whole homogenate. The homogenates were significant difference test. Histological results centrifuged at 18.000 × g and 4°C for 30 min were compared using Kruskal-Wallis variance to determine malondialdehyde (MDA) and analysis. When differences among the groups reduced glutathione (GSH) concentrations and were detected, group means were compared the superoxide dismutase (SOD) and catalase using the Mann-Whitney U-test. (CAT) activities or at 25.000 × g for 50 min to determine the glutathione peroxidase (GPx) ■ Results activity. Histopathological examinations Biochemical results For light microscopic evaluation, Antioxidant (SOD, CAT, GPx, and GSH) ovarian tissues were fixed in 10% formalin and oxidant parameters (MDA) in rat uterus and embedded in paraffin wax. Paraffin wax- are presented in Table 1. MDA levels were embedded specimens were cut into five μm significantly decreased, whereas GSH levels, 501 The therapeutic effects of curcumin and capsaicin against cyclophosphamide Acta Cir Bras. 2018;33(6):499-507 side effects on the uterus in rats Yilmaz E et al. GPx, CAT, and SOD activities were significantly the CYC+CPS groups compared with the control increased in uterine tissues in the CYC+CRC and group. Table 1 - MDA, GSH, SOD, GPx and CAT levels in tissues of the control group and the study group. Groups MDA GSH SOD GPx CAT Control 5.97±0.57a 44.0±2.02a 114.9±4.84a 495.5±45.8a 0.031±0.002a