Diabetik Ve Non-Diabetik Fare Kalbinde Oksitosin Reseptörünün İmmunohistokimyasal Lokalizasyonunun İncelenmesi

Year 2023, , 6 - 11, 30.06.2023

Serkan Arıcı , Seyit Ali Bingöl

https://doi.org/10.47027/duvetfd.1223132

Abstract

Bu çalışmada, diyabetik ve non-diyabetik fare kalp dokusunda oksitosin reseptörünün (OTR) immunohistokimyasal lokalizasyonunun incelenmesi amaçlanmıştır. Çalışmada 18 adet Balb-c cinsi ergin (8-12 haftalık) erkek fare; kontrol, sham ve diyabet grubu olarak belirlenmiştir. Diyabet grubu intraperitoneal (i.p.) enjeksiyonla streptozotosin (STZ) verilerek oluşturulmuştur. Sham grubuna ise i.p. yolla sodyum sitrat çözeltisi uygulanmıştır. Kontrol grubundaki hayvanlara ise herhangi bir uygulama yapılmamıştır. STZ uygulandıktan 30 gün sonra farelere eter anestezi altında servikal dislokasyon yapılarak kalp dokuları alınmıştır. Alınan her bir kalp dokusu düşey olarak ikiye bölünerek rutin histolojik işlemlerden geçirilerek parafinde bloklanıp kesitler alınmıştır. Alınan kesitler histolojik olarak incenlenmek üzere Hematoksilen Eosin (H&E), Crossman’ın üçlü boyaması ve Periyodik Asit Schiff (PAS) yapıldı. Oksitosin reseptörünün (OTR) immunoreaktivitesi Avidin-Biotin-Peroksidaz kompleks (ABC) metodu uygulanarak belirlenmiştir. Müdahale sonrasında, grupların canlı ağırlıkları, kan glikoz seviyesi, doku ağırlıkları ve kalp doku örneklerinde oksitosin reseptörünün immunohistokimyasal lokalizasyonu ve dokunun histolojik yapısı karşılaştırılmıştır. Kalp dokusunun ağırlıkları gruplar arasında karşılaştırıldığında istatistiksel düzeyde anlamlı bir fark yoktur (p>0.05). Histolojik incelemeler sonucunda diyabet grubunda diğer gruplara kıyasla kalbin miyokard bölgesindeki hücrelerde daha fazla dejenerasyon olduğu tespit edildi. İmmunohistokimyasal incelemeler sonucunda, sham ve kontrol grupları arasında OTR’nin immunoreaktivite seviyesi açısından istatistiksel olarak anlamlı bir fark yoktur. Diyabet grubunda diğer gruplara kıyasla OTR’nin immunoreaktivite seviyesi hücre membranı, sitoplazma ve purkinje hücrelerinde istatistiksel olarak anlamlı düzeyde daha az iken endotel ve kapiller alanlarda fark saptanmamıştır. Bu çalışma sonuçları oksitosin reseptörü, diyabet ve kalp dokusu arasında önemli bir ilişkinin olduğunu göstermiştir. Sonuç olarak, diyabetin oksitosin reseptörü vasıtasıyla kardiovasküler sistem üzerinde etkili olabileceği düşünülmektedir (p<0.05).

Keywords

Diyabet, İmmünohistokimya, Kalp, Oksitosin, Oksitosin Reseptörü

Supporting Institution

Kafkas Üniversitesi Rektörlüğü Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

2016-TS-42

Thanks

We thank Dr. Serdar Yiğit, Dr. Ayşe Aydoğan, Dr. Engin Yalmancı and Dr. Yakup Zühtü Birinci.

Investigation of Immunohistochemical Localization of Oxytocin Receptor in Diabetic and Non-Diabetic Mouse Heart

Abstract

The aim of this study was to investigate the immunohistochemical localization of oxytocin receptor (OTR) in diabetic and non-diabetic mouse heart tissue. Eighteen male Balb-c adult (8-12 week) mice were used in the study. Animals were divided into three groups; control, sham and diabetes. The diabetes group was given STZ by intraperitoneally (i.p) injections and diabetes was induced. Sham group was again treated with sodium citrate solution by i.p. The animals in the control group did not receive any treatment. After 30 days of STZ application, mice were cervical dislocated under ether anesthesia and their heart tissues were removed. Each heart tissue was vertically divided into two parts and routine histological procedures were applied and then tissues were blocked in paraffin and sections were taken. For histological examination, Haematoxylin&Eosin (H&E), Crossman’s Triple staining and Periodic Acid Schiff (PAS) were applied to the sections. Immunoreactivity of OTR was determined by Avidin-Biotin-Peroxidase Complex (ABC) method. At the end of the study period; the body weight of the groups, blood glucose level, tissue weights and immunohistochemical localization of OTR in heart tissue samples and histological structure of tissue were compared. When weights of heart tissue were compared between the groups, there was no statistically significant difference between the groups (p>0.05). As a result of histological examinations, it was found that there was more degeneration in the cells in the myocardium of the heart in the diabetes group compared to the other groups. Immunohistochemical examinations showed that OTR showed similar immunoreactivity in sham and control groups. In the diabetic group, the immunoreactivity of OTR was similar in endothelial and capillary areas, but less in cell membrane, cytoplasm and purkinje cells. In conclusion, the results of this study showed that there is a significant relationship between the OTR, diabetes and heart tissue. As a result, it is thought that diabetes may have an effect on the cardiovascular system through the OTR (p<0.05).

Keywords

diabet, heart, Immunohistochemistry, Oxytocin, Ocytocin receptor

Project Number

2016-TS-42

References

• Çavuşoğlu T, Çiftçi ÖD, Çağıltay E, et al. (2017). Diyabetik Kardiyomiyopati Sıçan Modelinde Oksitosin Etkilerinin Histolojik ve Biyokimyasal Olarak Incelenmesi. Dicle Tıp Dergisi. 44(2): 135-143.
• Işık S, Delibaşı T, Berker D, Aydın Y, Güler S. (2009) Kalp Hastalıklarında Diyabet Yönetimi. Anadolu Kardiyoloji Dergisi. 9: 238-247.
• Özel CB, Arıkan H, Dağdelen S, et al. (2021). Investigation of Cardiovascular Disease Risk Factors Knowledge and Physical Activity Levels in Patients with Type 2 Diabetes. Journal of Exercise Therapy and Rehabilitation. 8(1): 99-105.
• Du Vigneaud V. (1956). Trail of Sulfur Research: From Insulin to Oxytocin. Science. 123: 967-974. Bjorkstrand E, Eriksson M, Uvna Moberg K. (1996). Evidence of Peripheral and a Central Effect of Oxytocin on Pancreatic Hormone Release in Rats. Neuroendocrinology. 63: 377-383,
• Wang P, Wang SC, Liu X. (2022). Neural Functions of Hypothalamic Oxytocin and its Regulation. American Society for Neurochemistry. 14: 1-33.
• Paquin J, Danalache BA, Jankowski M, Mccann SM, Gutkowska J. (2004). Oxytocin Induces Differentiation of P19 Embryonic Stem Cells to Cardiomyocytes. Proc Natl Acad Sci. 99: 9550- 9555.
• Jankowski M, Danalache B, Wang D, et al. (2004). Oxytocin in Cardiac Ontogeny. Proc. Natl. Acad. Sci. USA 101: 13074-13079.
• Houshmand F, Faghihi M, Zahediasl S. (2009). Biphasic Protective Effect of Oxytocin on Cardiac Ischemia/Reperfusion Injury in Anaesthetized Rats. Peptides. 30(12): 2301-2308.
• Ondrejcakova M, Ravingerova T, Bakos J, Pancza D, Jezova D. (2009). Oxytocin Exerts Protective Effects on in Vitro Myocardial Injury Induced by Ischemia and Reperfusion. Can J Physiol Pharmacol. 87: 137-142.
• Balazova L, Krskova K, Suskp M, Sısovsky V, Hlavacova N, Olszanecki R. (2016). Metabolic Effects of Subchronic Peripheral Oxytocin Administration in Lean and Obese Zucker Rats. J Physiol Pharmacol. 67:4531-4541.
• Gutkowska J, Jankowski M, Lambert C, Mukaddam-Daher S, Zingg HH, Mccann SM. (1997): Oxytocin Releases Atrial Natriuretic Peptide by Combining with Oxytocin Receptors in the Heart. Proc Natl Acad Sci. 94: 11704-11709.
• Biggs LM, Hammock EAD. (2022). Oxytocin via Oxytocin Receptor Excites Neurons in the Endopriform Nucleus of Juvenile Mice. Scientific Reports. 12(1): 11401.
• Mineo H, Ito M, Muto H, Kamita H. (1997). Effect of Oxytocin Arginine-Vasopressin and Lysine-Vasopressin on Insulin and Glucagon Secretion in Sheep. Res in Veterinary Science. 62: 105-110.
• Wallin LA, Fawcet CP, Rosenfeld CR. (1996). Oxytocin Stimulates Glucagon and Insulin Section in Fetal and Neonatal Sheep. Endoc.125: 2289-2296.
• Knudizon J. (1981). Acute Effect of Oxytocin and Vasopressin on Plasma Levels of Glucagon, Insulin and Glucose in Rabbits, Horm Metab Res. 15:103-106.
• Aitszuler N, Hamshire J. (1981). Oxytocin Infusion Increases Plasma Insulin and Glucagon Levels and Glucose Production and Uptake in the Dog. Diabetes. 30: 112-114.
• Bingöl SA, Kocamış H. (2010). The Gene Expression Profile by RT-PCR and Immunohistochemical Expression Pattern of Catalase in the Kidney Tissue of Both Healthy and Diabetic Mice. Kafkas Univ Vet Fak Derg. 16 (5): 825-834.
• Shaukat A, Hussain G, Irfan S, Ijaz MU, Anwar H. (2022). Therapeutic Potential of MgO and MnO Nanoparticles within the Context of Thyroid Profile and Pancreatic Histology in a Diabetic Rat Model. Dose-Response. 20(3): 1-11.
• Luna LG. (1968). Manual of Histologic Staining Methods of Armed Forces Institute of Pathology. 3th ed., pp. 222-226. Mc GrawHill Book Comp, New York.
• Hsu SM, Raine L, Fanger H. (1981). A Comparative Study of the Peroxidase-Antiperoxidase Method and an Avidin-Biotin Complex Method for Studying Polypeptide Hormones with Radioimmunoassay Antibodies. American Journal of Clinical Pathology. 75(5): 734-738.
• Grover JK, Vats V, Rathi SS, Dawar R. (2001). Traditional Indian Anti-Diabetic Plants Attenuate Progression of Renal Damage in Streptozotocin Induced Diabetic Mice. J Ethnopharmacol. 76(3): 233-238.
• Wada T, Furuichi K, Sakai N, et al. (2000). Upregulation of Monocyte Chemoattractant Protein-1 in Tubulointerstitial Lesions of Human Diabetic Nephropathy. Kidney Int. 58(4): 1492-1499.
• Yeğin SÇ, Mert N. (2013). Deneysel Olarak Diyabet Oluşturulmuş Sıçanlarda HbA1c, MDA, GSH-Px ve SOD Miktarlarının Tayini. YYU Veteriner Fakültesi Dergisi. 24: 51-54.
• Demir E, Yılmaz Ö. (2013). Streptozotosin Ile Tip-2 Diyabet Oluşturulan Sıçanlarda Çam Yağının Antihiperglisemik ve Bazı Biyokimyasal Parametrelere Etkisi. Marmara Fen Bilimleri Dergisi. 25(3): 140- 156.
• Koroglu P, Senturk GE, Yucel D, Ozakpinar OB, Uras F, Arbak S. (2014) The Effect of Exogenous Oxytocin on Streptozotocin (STZ)-Induced Diabetic Adult Rat Testes. Peptides. 63: 47-54.
• Harackova M, Murphy MG. (1998). Effects of Chronic Diabetes Mellitus on the Electrical and Contractile Activities, 45Ca2+ Transport, Fatty Acid Profiles and Ultrastructure of Isolated Rat Ventricular Myocytes. Pflugers Arc. 411: 564-572.
• Cai F. (1989). Studies of Enzyme Histochemistry and Ultrastructure of the Myocardium in Rats with Streptozotocin Induced Diabetes. Zhonghua Yixue Za Zahi. 69(5): 276-278.
• Sun S, Dawuti A, Gong Difei. (2022). Puerarin-V Improve Mitochondrial Respiration and Cardiac Function in a Rat Model of Diabetic Cardiomyopathy via Inhibiting Pyroptosis Pathway through P2X7 Receptors. Int J Mol Sci. 23(21): 13015.
• Çetin A, Vardı N, Orman D. (2013). Deneysel Diyabetin Sıçan Kalp Dokusunda Meydana Getirdiği Histolojik Değişiklikler Üzerine Aminoguanidinin İyileştirici Etkileri. Sağlık Hizmetleri Meslek Yüksekokulu Dergisi. 4(1): 1–11.
• Wigger DC, Gröger N, Lesse A. (2020). Maternal Separation Induces Long-Term Alterations in the Cardiac Oxytocin Receptor and Cystathionine γ-Lyase Expression in Mice. Oxidative Medicine and Cellular Longevity. 2020: 1-10.
• Merz T, Denoix N, Wigger D. (2020). The Role of Glucocorticoid Receptor and Oxytocin Receptor in the Septic Heart in a Clinically Relevant, Resuscitated Porcine Model with Underlying Atherosclerosis. Frontiers in Endocrinology. 11: 1-8.
• Liu J, Liang Y, Jiang X, et al. (2021). Maternal Diabetes-Induced Suppression of Oxytocin Receptor Contributes to Social Deficits in Offspring. Frontiers in Neuroscience. 15: 634781.