{"paper_id":"7a7fa7db-ec49-47d5-96b1-a47b67643a1d","body_text":"Effect of Lycopene against Ovarian Torsion  Yilmaz et al.\nTHIEME\n32\nProtective Effect of Lycopene against Reperfusion \nInjury in Rats with Ovarian Torsion: A Biochemical \nand Histopathological Evaluation\nEmsal Pinar Topdagi Yilmaz 1 Harun Un2 Betul Gundogdu 3 Elif Polat 4 Seda Askin 4  \nYunus Emre Topdagi 5 Zekai Halici 6\n1Department of Gynecology and Obstetrics, Faculty of Medicine, \nAtaturk University, Erzurum, Turkey\n2Department of Biochemistry, Faculty of Pharmacy, Ağrı Ibrahim \nCecen University, Agri, Turkey\n3Department of Pathology, Faculty of Medicine, Ataturk University, \nErzurum, Turkey\n4Department of Biochemistry, Faculty of Medicine, Ataturk \nUniversity, Erzurum, Turkey\n5Department of Gynecology and Obstetrics, Faculty of Medicine, \nSanko University, Gaziantep, Turkey\n6Department of Pharmacology, Faculty of Medicine, Ataturk \nUniversity, Erzurum, Turkey\nAddress for correspondence   Yunus Emre Topdagi, MD, \nDepartment of Gynecology and Obstetrics, Faculty of Medicine, \nSanko University, Gazimuhtar Paşa Bulvarı No, 36, Şehitkamil \nGaziantep 27090, Turkey (e-mail: emretopdagi@hotmail.com).\nObjective The aim of our study was to evaluate the effect of two different doses of \nlycopene, an antioxidant, on experimentally induced ovarian ischemia/reperfusion (IR) \ninjury in rat model.\nMaterials and Methods  Twenty-four female rats were randomly divided into four \ngroups: sham operation (group 1), 3-hour ischemia, 3-hour reperfusion (IR) (group 2), \nand IR + 100 mg/kg lycopene (PO) (group 3), IR + 200 mg/kg of lycopene (group 4). \nThe rats’ superoxide dismutase (SOD), myeloperoxidase (MPO) activities, malondial -\ndehyde (MDA), and glutathione (GSH) levels were calculated. Ovarian tissue damage \nwas assessed using a histopathological scoring system.\nResults Serum parameter levels and histological scores showed that treatment with \nlycopene may be conservative approach to prevent IR injury after the ovarian detor -\nsion procedure.The improvement with lycopene was higher at 200 mg than at 100 mg. \nThe MPO and MDA values were significantly lower in groups 3 and 4 as compared \nwith group 2 ( p < 0.05), whereas the MPO and MDA values were lower in group 4 as \ncompared with group 3.The SOD and GSH values were significantly higher in groups \n3 and 4 as compared with group 2 ( p < 0.05), whereas the SOD and GSH values were \nhigher in group 4 as compared with group 3.Tissue damage scores were elevated in the \nIR group compared with the sham group, but the treatment with different lycopene \ndoses after reperfusion improved the histopathological tissue damage scores.\nConclusion  The results showed that lycopene treatment reduced ovarian IR dam -\nage. Antioxidant activity was found to increase in a dose-dependent manner. Lycopene \ntreatment may be conservative approach for ovarian torsion patients after the detor -\nsion procedure to prevent IR damage.\nAbstract\nKeywords\n ► ischemia/reperfusion\n ► lycopene\n ► ovarian torsion\n ► oxidative stress\nDOI https://doi.org/  \n10.1055/s-0040-1715553  \nISSN 0974-2727 .\n©2020 by The Indian Association \nof Laboratory Physicians\nJ Lab Physicians:2020;12:32–37\nOriginal Article\nArticle published online: 2020-08-11\n\n33\nEffect of Lycopene against Ovarian Torsion  Y ilmaz et al.\nJournal of Laboratory Physicians   Vol. 12   No. 1/2020\nIntroduction\nOvarian torsion is a gynecological emergency among women \nof reproductive age. Early diagnosis and management are \nimportant for the preservation of ovarian function. In cases \nof ovarian torsion, an ischemia/reperfusion (IR) injury may \ndevelop due to the release of free radicals and reactive oxy -\ngen species (ROS) during the detorsion process. 1 Oxidative \ntrauma occurs in a cell when the concentration of gener -\nated ROS exceeds that cell’s antioxidant capability.2,3 Various \nantioxidants have been used for the prevention of oxidative \ninjury and inflammation in ovaries subject to IR injury.\n4 \nMalondialdehyde (MDA) is the basic product of lipid peroxi-\ndation and is used to determine oxidative stress levels. 5 It is \nwell known that glutathione (GSH) is one of the most import-\nant indicators of the body’s antioxidant capacity.1 Lycopene, \na type of carotenoid, has antioxidant and chemopreventive \nproperties.\n3 In our study, rats with ovarian torsion–detorsion \ndamage were evaluated for the effects of lycopene. Moreover, \nthe study evaluated the degree of ovarian tissue damage by \nhistopathological examination and biochemically assessed \nthe levels of MDA, GSH, myeloperoxidase (MPO), and super -\noxide dismutase (SOD) enzyme activity.\nMaterials and Methods\nApproval from the Atatürk University Ethical Committee \nwas obtained before the study (ethical approval date: \n06.28.2018/07). The study was performed in the Animal \nLaboratory and Experimental Research Center of Atatürk \nUniversity in July 2018. Animals were treated in accor-\ndance with the Guide for the Care and Use of Laboratory \nAnimals (8th Edition, National Academies Press). In this \nstudy, 24 healthy, adult, nonpregnant, female Wistar Albino \nrats weighing between 228 and 272 g were used. The ani -\nmals were fed and kept in a cage at a constant room tem -\nperature and humidity under standard 12-hour light and \ndark laboratory conditions. All experimental processes were \nperformed when the rats were in the estrous phase. The rats \nwere randomly divided into four groups of six animals: sham \noperation (group 1) 1 mL of corn oil was applied by gavage \nmethod; IR group 2: IR + 100 mg/kg lycopene, group 3: IR + \n200 mg/kg lycopene, and group 4: lycopene 10% FS (Redivivo, \nDSM Nutritional Products). Lycopene was suspended in corn \noil and administered by gavage at doses of 100 and 200 mg \nper kg half an hour before the end of ischemia (2.5  hours of \nischemia).6\nSurgical Protocol\nAll procedures were performed under general anesthesia and \nsterile conditions. Each rat was intramuscularly injected with \n45 mg/kg of ketamine hydrochloride (Ketalar) and 5 mg/kg of \nxylazine hydrochloride (Rompun, Bayer) for general anesthe-\nsia. The skin of the abdomen was shaved and cleaned with \n10% povidone iodine. The lower abdomen was opened with \na 2-cm midline incision, and the bilateral ovary and adnexa \nwere exposed. The uterine horn and ovaries were specified. \nVascular clamps were placed just beneath the ovaries and over \nthe uterine horns. The incisions were closed with 4–0 silk \nsutures. The IR procedures were performed by one identical \nperson using the same technique. The adnexa were rotated \n720 degrees clockwise and fixed to the abdominal wall for \n3 hours w ith a 5–0 polydioxanone suture. In the detorsion \ngroups, the abdomen was reopened, and after the removal of \nthe fixation sutures, the ovaries were brought to their former \npositions with detorsion. To protect the rats from hypother-\nmia, the operating table was heated with a lamp from above \nand a heater from below. During the waiting period, the inci-\nsion line on the abdominal region was closed with a 3–0 silk \nsuture. In the sham group, a laparotomy was performed and \nthe incision was closed with a 3–0 nylon suture. A relaparot-\nomy was performed after a 3-hour period of all procedures \nin all groups, a bilateral oophorectomy was performed, and \nthe animals were sacrificed after their blood was taken. In \nall groups, ovarian tissue was kept in 10% formaldehyde for \nhistopathological evaluation and biochemical analyses.\nBiochemical Assay\nFor the biochemical analyses, the ovarian tissues were dis -\nsected out and frozen immediately at–80°C. Then, 0.1 g of \nthe tissue was homogenized with 900 µL of ice-cold phos -\nphate-buffered saline at pH 7.4 (10% w/v) and centrifuged \nat 4,000 rpm for 15  minut es at 4°C. The supernatants were \nstored at–80°C to analyze the SOD, MPO, MDA, and GSH \nlevels. The protein content of the supernatants was deter -\nmined by the Bradford method using bovine serum albumin \nas standard.\n7 The MDA levels, an index of lipid peroxida -\ntion, were determined through thiobarbituric acid reaction \nusing the method described by Ohkawa et al. 8 The product \nwas evaluated spectrophotometrically at 532 nm, and the \nresults are expressed as nmol/mg protein. MPO activity in \nthe supernatant was measured with the method described \nby Bradley et al.\n9 Then, 0.1 mL of the supernatant was mixed \nwith 2.9 mL of 50-mM phosphate buffer, pH 6.0, containing \n0.167 mg/mL of O-dianisidine dihydrochloride and 0.0005% \nhydrogen peroxide. The change in absorbance at 460 nm \nwas assayed spectrophotometrically. MPO enzyme activity \nwas recorded as U/mg protein in the sample. The GSH and \nSOD levels in the supernatants were measured with ELISA \n(enzyme-linked immunosorbent assay) methods using com-\nmercial kits (Cayman, cat no: 703002, and Cayman, cat no: \n706002, respectively).\nHistopathological Examination\nThe ovaries were fixed in 10% formaldehyde for 72  hours , \ndehydrated in a graded alcohol series, embedded in paraf -\nfin wax, and sectioned using a Leica RM2125RT microtome \n(Leica Microsystems). Five-mm-thick sections were used in \nthis study for histopathological examinations and evalua-\ntions. After placing the 5-mm-thick sections of tissue onto \nslides and after deparaffinization and rehydration, one sec -\ntion from each rat was stained with hematoxylin–eosin. All \nsections were examined and photographed using a light \nphotomicroscope (Olympus BX51 light microscope). Tissue \ndamage was histopathologically assessed for hemorrhage, \nvascular congestion, and polymorphonuclear leukocyte \n\n\n34\nJournal of Laboratory Physicians   Vol. 12   No. 1/2020\nEffect of Lycopene against Ovarian Torsion  Yilmaz et al.\n(PMNL) infiltration. At least five microscopic regions were \nexamined to score the specimens semiquantitatively. Each \nsample was scored for each criterion using a scale ranging \nfrom 0 to 3 (0, none; 1, mild; 2, moderate; 3, severe). Total \nscores were calculated from these parameters. One pathol -\nogist examined all the ovarian sections in a blinded fashion.\nStatistical Analysis\nThe data regarding the biochemical SOD, GSH, MDA, and MPO \nlevels were subjected to a one-way analysis of variance using \nthe IBM SPSS Statistics Version 20.0 (IBM Corp.). Differences \namong the groups were determined using the Duncan mul -\ntiple comparison test and were considered to be significant \nwhen the p-values were less than 0.05. All the results were \nexpressed as a mean standard deviation of the mean.\nResults\nBiochemical Results of Ovary Tissues\nIn this study, the antioxidant (SOD and GSH) and oxidant \n(MDA and MPO) parameters in the ovarian IR injury model \nin rats among different treatment groups are presented in \n►Fig. 1 . The MDA levels and MPO activity were significantly \ndecreased, whereas the GSH levels and SOD activities were \nsignificantly increased in the ovarian tissues in the IR + lyco-\npene groups compared with the IR group (p < 0.05). The SOD \nactivity and GSH levels were found to be lower and the MDA \nlevels and MPO activities were found to be higher in the IR \ngroup when compared with the IR + 100 mg and IR + 200 \nmg groups. Lycopene administration was found to amelio -\nrate SOD activity and GSH levels in a dose-dependent man -\nner. Furthermore, there were significant differences between \nthe IR + 100 mg and IR + 200 mg groups for all antioxidant \nand oxidant parameters (►Fig. 1) (p < 0.05). The SOD activity \nand GSH levels were found to be lower in the IR + 100 mg \nlycopene group as compared with the IR + 200 mg lycopene \ngroup. The MDA levels and MPO activities were found to be \nhigher in the IR + 100 mg lycopene group as compared with \nthe IR + 200 mg lycopene group. These results demonstrate \nan inverse correlation between lycopene levels and oxidative \nstress, supporting the hypothesis that lycopene supplemen -\ntation could ameliorate oxidative stress in the rat ovary. Thus, \nin this study, the lowest SOD and GSH levels were observed \nin group 2. However, lycopene given before the reperfusion \nperiod increased SOD activity. The most dramatic increase \noccurred in group 4, which received the 200 mg/kg dose. \nThe 200 mg/kg dose of lycopene was more effective than the \n100 mg/kg dose.\nHistopathological Results\nAll rats were evaluated for tissue damage by assessing param-\neters such as congestion, PMNL infiltration, and hemorrhage. \nThe histopathological scores for all four groups are listed. \nThe ovarian histopathological changes for the IR, IR + lyco -\npene 100 mg, and IR + lycopene 200 mg groups are shown \nin ►Fig. 2 . The histopathological evaluation showed that tis-\nsue damage significantly increased in the IR group compared \nwith the lycopene treatment groups. The tissue damage in \nthe lycopene treatment groups significantly decreased com -\npared with the IR group. Cellular improvements on these \nhistopathological parameters as a result of lycopene can be \na reflection of its antioxidant effect. Vascular congestion, \nhemorrhage, and PMNL infiltration were significantly higher \nin the IR group than in the sham group. Congestion, hemor -\nrhage, and PMNL infiltration regressed to the degrees of mild \nand medium from severe in the IR group, which was given \nthe lycopene treatment. There were significant differences in \ntissue damage between the lycopene 100 mg and lycopene \n200 mg groups. The sham group (group 1) had a normal his -\ntological architecture with minimal congestion, including the \ncortex and medulla. The cortex consists of ovarian follicles \nand corpus luteum, whereas the medulla consists of vascular \nFig. 1  Effects of lycopene treatments on SOD and MPO activities, and GSH and MDA levels in rat ovaries. Means in the same column by the \nsame letter are not significantly different to the Duncan test ( p < 0.05). Results are presented as means ± standard deviation. GSH, glutathione; \nIR, ischemia/reperfusion; MDA, malondialdehyde; MPO, myeloperoxidase; SHAM, sham-operated; SOD, superoxide dismutase.\n\n\n35\nEffect of Lycopene against Ovarian Torsion  Y ilmaz et al.\nJournal of Laboratory Physicians   Vol. 12   No. 1/2020\nstructures and connective tissue. The tunica albuginea that \nsurrounds the ovary had a normal appearance ( ►Fig. 2A ,B). \nIn the ovarian torsion IR group (group 2), intensive neutro -\nphil infiltration was observed in the peripheral connective \ntissue, and hemorrhage was observed between this tissue \nand the ovarian tissue ( ►Fig.  2C ). Significant congestion \nwas observed in the vascular structures in the medulla \n(►Fig. 2D ). The ovarian torsion + 100 mg/kg lycopene group \n(group 3) had focal hemorrhage compared with the IR group \n(►Fig. 2E). In the IR + lycopene 100 mg group, congestion and \ninflammatory cell density were lower than in the IR group \n(►Fig.  2F,G). Hemorrhage was not observed in the ovarian \ntorsion + 200 mg/kg lycopene group (group 4) ( ►Fig.  2H). \nThe congestion and infiltration of infective cells were mini -\nmal (►Fig. 2J ,K) in all groups.\nDiscussion\nOvarian torsion frequently occurs in women during the pre -\nmenarchal or reproductive years. For that reason, early and \naccurate diagnosis is very important for a woman’s reproduc-\ntive health.10 Ovarian torsion must be diagnosed and treated \nto prevent potential necrosis leading to infertility.11 Detorsion \nmight be considered to restore the ovarian blood flow. \nHowever, reperfusion might lead to more serious injury, 12 \nknown as an IR injury.13 In oxidative stress, there is an imbal-\nance between the production and elimination of ROS.3 The IR \ncondition oxidizes cellular membrane lipids and leads to the \nformation of toxic products such as MDA.14 On the other hand, \ncellular protection against oxidative damage is provided by \nantioxidant enzymes and nonenzymatic compounds such as \nGSH or SOD.4,11 MPO, a major component of neutrophil azuro-\nphilic granules, is often released from stimulated PMNLs at \ninflammation sites and is involved in the generation of ROS.15 \nMDA is the end product of lipid peroxidation, which induces \nischemic injury, whereas GSH is the most significant cellu -\nlar antioxidant compound. 16 It defends cells against oxidant \ndamage by entering into reactions with GSH and free rad -\nicals.11 SOD and MPO are antioxidant enzyme components \nof the defense mechanism against the activities of oxidative \nsubstances. Increased levels of these enzymes protect the tis-\nsue during ovarian IR damage. 5,17,18 After ovarian detorsion, \nto protect the ovarian reserve against IR damage, prophylac-\ntic measures are necessary.19 Therefore, several studies have \nfocused on pharmaceutical agents with antioxidant effects to \nprevent ovarian IR damage in animal models.1,3,5,18\nThe ischemia that develops due to torsion has been \nreported to increase the blood levels of lipid peroxidation \nproducts.20 It is known that antioxidant treatment helps pre-\nvent tissue damage related to increases in oxidant production. \nAntioxidant agents reduce oxidized biomolecules and repair \nexisting oxidative injury. 21 Therefore, to prevent ischemic \ninjury in ovarian torsion, many prophylactic agents such as \nerdosteine, selenium, erythropoietin, vardenafil, vitamin C, \nand curcumin have been used before and after ischemia.22,23\nCellular and molecular studies have revealed lycopene, \nwhich not only inhibits lipid peroxidation but also quenches \nROS,24 to be one of the strongest antioxidants. 25 It has been \ncommonly used as an antioxidant agent in traditional medi -\ncine and has been reported to have protective effects for the \ntreatment of cardiovascular diseases, neurotoxicity, hepatic \ninjury, and nephrotoxicity.26\nAcetaminophen overdoses are causes of hepatic necrosis \nand acute liver failure. 27 It has been reported that lycopene \ndefends against acetaminophen-induced liver injury in \nmice by increasing antioxidant substances including GSH. 28 \nCarbon tetrachloride (CCl4) causes liver injury in experimen-\ntal studies. 29 In the study by Pinto et al, it was shown that \nlycopene acted as a therapeutic agent against CCl 4-induced \nacute liver damage in experimental animals. A significant \nFig. 2  Effects of lycopene treatments on light micrographs in rats’ ovaries (hematoxylin–eosin (H&E) staining). ( A) Sham group (group 1) \nwith minimal congestion, x400, H&E. ( B) Sham group, (group 1) with no inflammation and hemorrhage, x400, H&E. ( C) Ischemia/reperfusion \n(IR) group (group 2) with hemorrhage (unidirectional arrow) and inflammation (bidirectional arrow), x400, H&E. ( D) IR group (group 2) with \ncongestion (arrow) x400, H&E. ( E) IR+ lycopene 100 mg/kg (group 3) with focal hemorrhage, x400, H&E. ( F) IR+ lycopene 100 mg/kg (group 3) \nwith congestion, x400, H&E. ( G) IR+ lycopene 100 mg/kg (group 3) with inflammation (arrow), x400, H&E. ( H) IR+ lycopene 200 mg/kg (group \n4) with hemorrhage not detected, x400, H&E. ( J) IR + lycopene 200 mg/kg (group 4) with minimal congestion, x400, H&E. ( K) IR+ lycopene \n200 mg/kg (group 4) with minimal inflammation, x400, H&E.\n\n\n36\nJournal of Laboratory Physicians   Vol. 12   No. 1/2020\nEffect of Lycopene against Ovarian Torsion  Yilmaz et al.\nincrease was observed in the GSH concentration and SOD \nactivity in lycopene-treated groups. 30 Previous studies have \nreported that gentamicin nephrotoxicity is related to oxida -\ntive stress and hydroxyl radicals.31 Studies on the prevention \nof gentamicin toxicity reported that treatment with lyco -\npene markedly increased antioxidant levels and reduced \nthe urea and creatinine levels in rats given gentamicin. 32 \nCyclosporine A is used as an immunosuppressive drug after \ntransplantation, but it has nephrotoxicity side effects. 33 It \nwas stated that giving lycopene to rats caused a decrease in \nthe urea and creatinine serum concentrations, an increase \nin SOD, and the suppression of MDA. 34 A study by Liu et al \nreported that lycopene ameliorates ovarian aging in chick -\nens. The ovarian tissues of young and old hens were treated \nwith lycopene to verify its protective effects. Treatment \nwith lycopene reported a decrease in the MDA content and \nROS levels in both young and old ovarian tissues. 35\nMany studies have shown that MDA levels increased \nrelated IR injuries.36 It has been stated that this increase is a \nclear sign of an IR injury. 37 In our study, the histopathologi -\ncal and biochemical evaluations showed that 100 or 200 mg \ndoses of lycopene were effective in reducing ovarian tissue \ndamage. Furthermore, when compared with the sham oper -\nated group, the increase in the MDA level in the IR group \nwas significant. On the other hand, the IR process caused a \nsignificant decrease in the GSH level in the ovarian tissue of \nrats compared with the sham group. Lycopene significantly \nimproved the GSH level in the lycopene treatment groups \nwhen compared with the IR group. The SOD activity was sup-\npressed in the IR group compared with the sham group. But \na significant increase was found in the lycopene treatment \ngroups compared with the IR group. On the contrary, MPO \nactivity was significantly increased in the IR group when \ncompared with the sham group, and when lycopene was sup-\nplied, MPO activity significantly decreased. Consistent with \nour results, Tok et al reported that the levels of oxidant indi-\ncators such as MDA and MPO increased and the levels of anti-\noxidant indicators such as GSH decreased in the IR group.38\nOn the contrary, Sayar at al showed that a significant rise \nin the SOD enzyme activity was detected in the IR group, \nwhereas a significant reduction in the levels of this marker \nwas observed when antioxidant material was applied to an \nIR injury.39\nHistopathologically, dilated congested blood vessels, \ninflammation, and severe hemorrhage in ovarian tissue with \nthe oxidative trauma caused by IR were also observed in \nthis study. Confirming our results, similar histopathological \nchanges in rat ovaries related to IR injury were reported in \nprevious experimental studies.1,40\nAn increase in antioxidant enzyme levels is a reflection of \nincreased oxidative stress. In addition, it was detected that \nenzyme levels decreased in the rats induced with an IR when \nadministered lycopene. These results are also important in \nterms of the antioxidant effects of lycopene that might be \na result of toxic agents resulting from its administration. In \nour study, the histopathological and biochemical evalua -\ntions showed that lycopene was effective in reducing ovarian \ntissue damage. It was observed that the dose-dependent \nactivity increased.\nIn conclusion, the results of this study showed that lyco -\npene has protective effects on oxidative stress induced by \nischemia in ovarian torsion and increases the dose-depen -\ndent effect. However, further clinical studies are required to \nreach accurate results.\nAuthor Contribution\nP. T. Y. contributed to data collection/management and \nprotocol/project development. H. U. contributed to man -\nuscript writing/editing and data analysis. B. G. contrib -\nuted to data analysis. E. P. contributed to protocol/project \ndevelopment. S. A. contributed to protocol/project devel -\nopment. Y E. T. contributed to data collection/manage -\nment. Z. H. contributed to data analysis and protocol/\nproject development.\nFunding\nThe authors declared that this study received no financial \nsupport.\nEthics Committee Approval\nEthics committee approval was received for this study \nfrom the ethics committee of Ataturk University. Animals \nwere treated in accordance with the Guide for the Care \nand Use of Laboratory Animals (8th Edition, National \nAcademies Press).\nConflict of Interest\nNo conflict of interest was declared by the authors.\nReferences\n1 Bayir Y, Cadirci E, Polat B, et al. Aliskiren - a promising strategy \nfor ovarian ischemia/reperfusion injury protection in rats via \nRAAS. Gynecol Endocrinol 2016;32(8):675–683\n2 Oral A, Halici Z, Bayir Y, et al. Effects of oral zinc administra -\ntion on long-term ipsilateral and contralateral testes damage \nafter experimental testis ischaemia-reperfusion. Andrologia \n2017;49(6):e12673\n3 Sahin K, Yenice E, Tuzcu M, et al. Lycopene protects against \nspontaneous ovarian cancer formation in laying hens. J Cancer \nPrev 2018;23(1):25–36\n4 Oral A, Odabasoglu F, Halici Z, et al. Protective effects of \nmontelukast on ischemia-reperfusion injury in rat ovaries \nsubjected to torsion and detorsion: biochemical and histo -\npathologic evaluation. Fertil Steril 2011;95(4):1360–1366\n5 Yigiter M, Halici Z, Odabasoglu F, et al. Growth hormone \nreduces tissue damage in rat ovaries subjected to torsion and \ndetorsion: biochemical and histopathologic evaluation. Eur J \nObstet Gynecol Reprod Biol 2011;157(1):94–100\n6 Cakir Gungor AN, Gencer M, Karaca T, et al. The effect of hes -\nperetin on ischemia-reperfusion injury in rat ovary. Arch \nGynecol Obstet 2014;290(4):763–769\n7 Bradford MM. A rapid and sensitive method for the quantita -\ntion of microgram quantities of protein utilizing the principle \nof protein-dye binding. Anal Biochem 1976;72:248–254\n8 Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in ani -\nmal tissues by thiobarbituric acid reaction. Anal Biochem \n1979;95(2):351–358\n9 Bradley PP, Priebat DA, Christensen RD, Rothstein G. \nMeasurement of cutaneous inflammation: estimation of \n\n\n37\nEffect of Lycopene against Ovarian Torsion  Yilmaz et al.\nJournal of Laboratory Physicians   Vol. 12   No. 1/2020\nneutrophil content with an enzyme marker. J Invest Dermatol \n1982;78(3):206–209\n10 Eser A, Hizli D, Haltas H, et al. Effects of curcumin on ovar -\nian ischemia-reperfusion injury in a rat model. Biomed Rep \n2015;3(6):807–813\n11 Halici Z, Karaca M, Keles ON, et al. Protective effects of \namlodipine on ischemia-reperfusion injury of rat ovary: \nbiochemical and histopathologic evaluation. Fertil Steril \n2008;90(6):2408–2415\n12 Zimmerman BJ, Granger DN. Reperfusion injury. Surg Clin \nNorth Am 1992;72(1):65–83\n13 Carden DL, Granger DN. Pathophysiology of ischaemia-reper -\nfusion injury. J Pathol 2000;190(3):255–266\n14 Un H, Bayir Y, Halici Z, et al. The effects of RAAS Inhibition in \nrate limiting step by Aliskiren on testicular torsion injury in \nrats. J Urol 2015;194(3):828–833\n15 Ece A, Kelekçi S, Hekimoğlu A, et al. Neutrophil activa -\ntion, protein oxidation and ceruloplasmin levels in chil -\ndren with Henoch-Schönlein purpura. Pediatr Nephrol \n2007;22(8):1151–1157\n16 Kaya C, Karabulut R, Turkyilmaz Z, et al. Lycopene has \nreduced renal damage histopathologically and biochemically \nin experimental renal ischemia-reperfusion injury. Ren Fail \n2015;37(8):1390–1395\n17 Dogan C, Halici Z, Topcu A, et al. Effects of amlodipine on \nischaemia/reperfusion injury in the rat testis. Andrologia \n2016;48(4):441–452\n18 Cadirci E, Oral A, Odabasoglu F, et al. Atorvastatin reduces \ntissue damage in rat ovaries subjected to torsion and detor -\nsion: biochemical and histopathologic evaluation. Naunyn \nSchmiedebergs Arch Pharmacol 2010;381(5):455–466\n19 Halestrap AP, Clarke SJ, Javadov SA. Mitochondrial permeability \ntransition pore opening during myocardial reperfusion–a tar -\nget for cardioprotection. Cardiovasc Res 2004;61(3):372–385\n20 Akgür FM, Kilinç K, Aktuğ T. Reperfusion injury after detorsion \nof unilateral testicular torsion. Urol Res 1993;21(6):395–399\n21 Kisaoglu A, Borekci B, Yapca OE, Bilen H, Suleyman H. Tissue \ndamage and oxidant/antioxidant balance. Eurasian J Med \n2013;45(1):47–49\n22 Sağsöz N, Kisa U, Apan A. Ischaemia-reperfusion injury of rat \novary and the effects of vitamin C, mannitol and verapamil. \nHum Reprod 2002;17(11):2972–2976\n23 Yildirim N, Simsek D, Kose S, et al. The protective effect of \nGingko biloba in a rat model of ovarian ischemia/reperfusion \ninjury: improvement in histological and biochemical parame -\nters. Adv Clin Exp Med 2018;27(5):591–597\n24 Kawata A, Murakami Y, Suzuki S, Fujisawa S. Anti-inflammatory \nactivity ofβ-carotene, lycopene and tri- n-butylborane, a scav-\nenger of reactive oxygen species. In Vivo 2018;32(2):255–264\n25 Basu A, Imrhan V. Tomatoes versus lycopene in oxidative stress \nand carcinogenesis: conclusions from clinical trials. Eur J Clin \nNutr 2007;61(3):295–303\n26 Hedayati N, Naeini MB, Nezami A, et al. Protective effect \nof lycopene against chemical and natural toxins: a review. \nBiofactors 2019;45(1):5–23\n27 Larson AM, Polson J, Fontana RJ, et al; Acute Liver Failure Study \nGroup. Acetaminophen-induced acute liver failure: results of \na United States multicenter, prospective study. Hepatology \n2005;42(6):1364–1372\n28 Bandeira ACB, da Silva RC, Rossoni JV, et al. Lycopene pretreat-\nment improves hepatotoxicity induced by acetaminophen in \nC57BL/6 mice. Bioorg Med Chem 2017;25(3):1057–1065\n29 Brattin WJ, Glende EA Jr, Recknagel RO. Pathological mecha -\nnisms in carbon tetrachloride hepatotoxicity. J Free Radic Biol \nMed 1985;1(1):27–38\n30 Pinto C, Rodriguez-Galdon B, Cestero JJ, Macias P. \nHepatoprotective effects of lycopene against carbon tetra -\nchloride-induced acute liver injury in rats. J Funct Foods \n2013;5(4):1601–1610\n31 Walker PD, Barri Y, Shah SV. Oxidant mechanisms in gentami -\ncin nephrotoxicity. Ren Fail 1999;21(3-4):433–442\n32 Karahan I, Ateşşahin A, Yilmaz S, Ceribaşi AO, Sakin F. Protective \neffect of lycopene on gentamicin-induced oxidative stress and \nnephrotoxicity in rats. Toxicology 2005;215(3):198–204\n33 Cohen DJ, Loertscher R, Rubin MF, Tilney NL, Carpenter CB, \nStrom TB. Cyclosporine: a new immunosuppressive agent for \norgan transplantation. Ann Intern Med 1984;101(5):667–682\n34 Ateşşahin A, Ceribaşi AO, Yilmaz S. Lycopene, a carot -\nenoid, attenuates cyclosporine-induced renal dysfunction \nand oxidative stress in rats. Basic Clin Pharmacol Toxicol \n2007;100(6):372–376\n35 Liu X, Lin X, Zhang S, et al. Lycopene ameliorates oxidative \nstress in the aging chicken ovary via activation of Nrf2/HO-1 \npathway. Aging (Albany NY) 2018;10(8):2016–2036\n36 Celik M, Aksoy AN, Aksoy H, Aksoy Y, Halici Z. Sildenafil \nreduces ischemia-reperfusion injury in rat ovary: biochem -\nical and histopathological evaluation. Gynecol Obstet Invest \n2014;78(3):162–167\n37 Erten SF, Kocak A, Ozdemir I, Aydemir S, Colak A, Reeder BS. \nProtective effect of melatonin on experimental spinal cord \nischemia. Spinal Cord 2003;41(10):533–538\n38 Tok A, Sener E, Albayrak A, et al. Effect of mirtazapine on oxi -\ndative stress created in rat kidneys by ischemia-reperfusion. \nRen Fail 2012;34(1):103–110\n39 Sayar I, Bicer S, Gursul C, Gürbüzel M, Peker K, Işik A. Protective \neffects of ellagic acid and ozone on rat ovaries with an ischemia/\nreperfusion injury. J Obstet Gynaecol Res 2016;42(1):52–58\n40 Yapca OE, Kumbasar S, Salman S, et al. Controlled reperfusion \nfor different durations in the treatment of ischemia-reperfu -\nsion injury of the rat ovary: evaluation of biochemical features, \nmolecular gene expression, and histopathology. Can J Physiol \nPharmacol 2015;93(4):269–274","source_license":"CC0","license_restricted":false}