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    Effect of vitamin B12 on methotrexate-induced cardiotoxicity in rats
    (Mashhad Univ Med Sciences, 2024) Kuloglu, Nurhan; Karabulut, Derya; Kaymak, Emin; Akin, Ali Tugrul; Ceylan, Tayfun; Yildirim, Aysegul Burcin; Yakan, Birkan
    Objective(s): Methotrexate (MTX) is a drug with anti-inflammatory and immunosuppressive effects and is also a folic acid antagonist. Our aim in this study is to determine the molecular mechanisms of cardiotoxicity caused by MTX, a chemotherapeutic drug, and to evaluate the protective effects of vitamin B12 on this toxicity. Materials and Methods: A total of 32 rats were used in our study and 4 groups were formed. Control group, Vit B12 group (3 mu g/kg B12 for 15 days, IP), MTX group (20 mg/kg MTX single dose on day 8 of the experiment, IP), MTX +Vit B12 group (3 mu g/kg, IP ), Vit B12 throughout the 15 days, and a single dose of 20 mg/kg MTX (IP) on day 8 of the experiment. Immunohistochemically, expressions of hypoxia-inducible factor 1 alpha (HIF1-alpha), vascular endothelial growth factor receptor -2 (VEGFR-2), erythropoietin (EPO), and interleukin-6 (IL -6) were evaluated in the heart tissue. Total catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) levels were measured in the heart tissue. At the same time, ANP and NT-proBNP levels were measured in the blood serum. Results: In the study, the expression of HIF1-alpha and VEGFR-2 increased significantly in the MTX group, while IL -6 and EPO significantly decreased. At the same time, CAT and SOD levels were significantly decreased and MDA levels increased significantly in the MTX group. While vitamin B12 significantly corrected all these values, it also greatly reduced the increases in ANP and NT-proBNP levels caused by MTX. Conclusion: It is important to use Vit B12 before and after MTX administration to replace the folate that MTX has reduced.
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    Immunoreactivity of Kir3.1, muscarinic receptors 2 and 3 on the brainstem, vagus nerve and heart tissue under experimental demyelination
    (Pergamon-Elsevier Science Ltd, 2023) Akyuz, Enes; Doganyigit, Zueleyha; Okan, Asli; Yilmaz, Seher; Ucar, Sumeyye; Akin, Ali Tugrul
    Aims: Demyelination affects the propogation of neuronal action potential by slowing down the progression. This process results in a neuro-impairment like Multiple Sclerosis (MS). Evidence show that MS also contributes to involvement of the autonomic system. In the molecular approach to this involvement, we aimed to observe muscarinic ACh receptor 2-3 (mAChR2-3), and inwardly rectifying potassium channel 3.1 (Kir3.1) immunore-activities on the brainstem, vagus nerve, and heart under cuprizone model.Main methods: Wistar albino rats were randomly divided into 8 groups; duplicating 4 groups as male and female: control groups (n = 3 +3), Cuprizone groups (n = 12 +12), sham groups (n = 4 +4), and carboxy-methyl-cellulose groups (n = 3 +3). Cuprizone-fed rats underwent demyelination via Luxol fast blue (LFB) staining of the hippocampus (Gyrus dentatus and Cornu Ammonis) and cortex. Immunohistochemistry analysis followed to the pathologic measurement of the brainstem, vagus nerve, and heart for mAChR2, mAChR3 and Kir3.1 proteinsKey findings: A significant demyelination was observed in the hippocampus and cortex tissues of rats in the female and male cuprizone groups. Myelin basic protein immunoreactivity demonstrated that cuprizone groups, in both males and females, had down-regulation in the hippocampus and cortex areas. The weights of the cuprizone-fed rats significantly decreased over six weeks. Dilated blood vessels and neuronal degeneration were severe in the hippocampus and cortex of the cuprizone groups. In the female cuprizone group, expression of mAChR2 and mAChR2 was significantly increased in the brainstem, atrium/ventricle of heart, and left/right sections of vagus nerve. Kir3.1 channels were also up-regulated in the left vagus nerve and heart sections of the female cuprizone groupSignificance: Especially in our data where female-based significant results were obtained reveal that demyelin-ation may lead to significant mAChR2, mAChR3 and Kir3.1 changes in brainstem, vagus nerve, and heart. A high immunoreactive response to demyelination at cholinergic centers may be a new target
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    In vivo Gonadoprotective Effects of Myricetin on Cisplatin- Induced Testicular Damage via Suppression of TLR4/NF-?B Inflammation Pathway and Heat-Shock Response
    (Soc Chilena Anatomia, 2023) Akin, Ali Tugrul
    The aim of this study is to reveal the gonadoprotective effects of myricetin (MYC), which has many biological properties, on cisplatin (CP)-induced testicular damage in rats. For this purpose, 40 male Wistar albino rats were divided into 4 groups as Control (group given no treatment), MYC (group given 5 mg/kg/i.p myricetin for 7 days), CP (group given 7 mg/kg/i.p cisplatin at 7th day) and MYC + CP (group given 5 mg/kg/i.p myricetin for 7 days before 7 mg/kg/i.p cisplatin injection). After administrations, testicular tissues of animals were extracted and processed according to tissue processing protocol. Hematoxylin & Eosin staining were performed to evaluate the histopathological changes and Johnsen'sTesticular Biopsy Score (JTBS) was applied and mean seminiferous tubule diameters (MSTD) were measured to compare experimental groups in terms of histopathological changes. Moreover, TLR4, NF-kappa B, HSP70 and HSP90 expression levels were detected by immunohistochemical staining and the density of immunoreactivity were measured to determine the difference in the expression levels of these factors among groups. Additionally, testicular apoptosis was detected via TUNEL assay. JTBS and MSTD data were significantly lower in CP group compared to other groups and MYC administrations significantly protects testicular tissue against CP-induced damage. Moreover, TLR4, NF-kappa B, HSP70 and HSP90 expressions and apoptotic cells significantly increased in the CP group (p<0.05). However, MYC administrations exerted a strong gonadoprotective effect on testicular tissue in terms of these parameters in MYC+CP group (p<0.05). According to our results, we suggested that MYC can be considered as a protective agent against cisplatin-induced testicular damage.
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    Protective effect of melatonin on cisplatin induced acute kidney injury: Role of regulation of heat shock proteins expressions
    (Natl Inst Science Communication-Niscair, 2023) Akin, Ali Tugrul; Unsal, Murat; Ceylan, Tayfun; Kaymak, Emin; Ozturk, Emel; Kuloglu, Nurhan; Karabulut, Derya
    Chemotherapy is one of the major treatment approaches for cancer, and these agents are known to cause severe side effects, including damaging vital organs. Cisplatin (CP) is a commonly used chemotherapeutic agent in the treatment of many cancer types, particularly lung, breast, ovarian, testicular and head-neck cancers. CP is reported to cause damage to damage on brain, kidney, liver and gonads. In this study, is we investigated the protective effects of melatonin (MEL) in acute kidney injury (AKI) induced by CP via assessment of heat-shock protein (HSP) induction in rats. For this purpose, total 40 Wistar albino rats were divided into four groups: Control (n=10), MEL (n=10, 10 mg/kg/i.p. melatonin for 8 days), CP (n=10, 7 mg/kg/i.p. cisplatin at the 5th day), and CP+MEL (n=10, 10 mg/kg/i.p. melatonin for 8 days and 7 mg/kg/i.p. cisplatin at the 5th day). After kidney tissues were extracted, histopathological changes were evaluated and immunoreactivities of HSP47, HSP60, HSP70 and HSP90 in renal cortex were detected via immunohistochemistry. Moreover, blood serum BUN (blood urea nitrogen), creatinine and uric acid levels were measured to assess kidney function. CP group showed histopathological deterioration, and MEL treatment attenuated this damage in CP+MEL group. An increase in HSPs immunoreactivities were detected in renal cortex of CP group when compared with the control and MEL groups, showing increased HSP response because of CP-induced AKI. However, CP-induced HSP induction was significantly lower in the CP+MEL group. Similarly, blood serum BUN, creatinine and uric acid levels were higher in CP group while CP+MEL group showed decreased levels of these parameters. Our results suggest that MEL could exert a significant protective effect against CP-induced AKI via reducing HSP response.
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    Therapeutic effect of thymoquinone on brain damage caused by nonylphenol exposure in rats
    (Wiley, 2023) Ceylan, Tayfun; Akin, Ali Tugrul; Karabulut, Derya; Tan, Fazile Canturk; Taskiran, Mehmet; Yakan, Birkan
    Nonylphenol (NP), causes various harmful effects such as cognitive impairment and neurotoxicity. Thymoquinone (TQ), has antioxidant, anti-inflammatory, and neuroprotective properties. In this study, our aim is to investigate the effects of TQ on the brain damage caused by NP. Corn oil was applied to the control group. NP (100 mg/kg/day) was administered to the NP and NP + TQ groups for 21 days. TQ (5 mg/kg/day) was administered to the NP + TQ and TQ groups for 7 after 21 days. At the end of the experiment, the new object recognition test was applied to the rats and the rats were killed and their brain tissues were removed. Sections taken from brain tissues were stained with hematoxylin-eosin for histopathological evaluation. In addition, neuronal nuclei (NeuN), glial fibrillary acidic protein (GFAP), Cas-3, and nerve growth factor (NGF) immunoreactivities were evaluated in brain tissue sections. In addition, malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) activities were determined. Comet assay was applied to determine DNA damage in cells. The results of our study showed that NP, caused behavioral disorders and damage to the cerebral cortex in rats. This damage in the form of neuron degeneration seen in the cortex was associated with apoptosis involving Cas-3 activation, increased DNA damage, and free oxygen radicals. NP, SOD, and CAT caused a decrease in enzyme activities. In addition, the cellular protein NeuN was decreased, astrocytosis-associated GFAP was increased, and growth factor NGF was decreased. When all our evaluations are taken together, treatment with TQ showed an ameliorative effect on the behavioral impairment and brain damage caused by NP exposure.
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    Thymoquinone reduces methotrexate-induced heart damage: a histopathological study in rats
    (Cukurova Univ, Fac Medicine, 2023) Yildirim, Aysegul Burcin; Kaymak, Emin; Ceylan, Tayfun; Akin, Ali Tugrul; Kuloglu, Nurhan; Sayan, Meryem; Deger, Necla
    Purpose: The study aimed to evaluate the effect of thymoquinone on cardiac tissue in MTX-induced cardiac toxicity in rats with various parameters.Materials and Methods: Group I (n=8) was administered intraperitoneal saline for 10 days. Intraperitoneal olive oil was applied to Group II (n=8) for 10 days. Group III (n=8) received 10 mg/kg Thymoquinone (THQ) intraperitoneally for 10 days. Group IV (n=8) was administered a single dose of 20 mg/kg Methotrexate (MTX), 500 mg/20 ml, intraperitoneally on the 1st day of the experiment. Group V (n=8) MTX: 20 mg/kg single dose intraperitoneally on the 1st day; THQ: 10mg/kg i.p. administered for 10 days. Since Methotrexate was in liquid form, no solvent was used. At the end of the experimental period, the rats were sacrificed for analysis of heart tissue. The structure of heart tissue was evaluated by hematoxylin-eosin staining. Immunohistochemically, Connexin-43, HSP90, and HIF- 1 alpha antibodies were stained.Results: Group IV was found to have histopathological deterioration, which was ameliorated by THQ. In addition to this; Connexin-43 immunoreactivity was the lowest in Group IV compared to other groups: 108.5 +/- 7.4. Compared to other groups, HSP90 immunoreactivity was highest in Group IV: 103.6 +/- 10.4. Compared to other groups, HIF-1 alpha immunoreactivity was highest in Group IV: 95.2 +/- 9.1.Conclusion: Thymoquinone has a positive effect on Connexin-43, one of the proteins providing conduction in intercalary discs, HSP90, one of the chaperones in the cell and HIF-1 alpha expression against MTX toxicity. At the same time, THQ provides a significant improvement in cardiac tissue histopathologically by showing a cardioprotective effect.