Investigation of the Effects of Streptozotocin Applied in Different Doses on Rat Liver with Stereological Methods

Yıl 2024, Cilt: 35 Sayı: 1, 53 - 58, 29.03.2024

Saadet Yıldız Murat Çetin Rağbetli Yasin Tülüce

https://doi.org/10.36483/vanvetj.1362601

Öz

Diabetes mellitus is a group of chronic metabolic conditions, all of which are characterized by elevated blood glucose levels resulting from the body's inability to produce insulin or resistance to insulin action, or both. Insulin is the main hormone that regulates glucose production and use by body tissues and organs. Diabetes affects all systems in the body, including the liver. Streptozotocin is an antibiotic that performs pancreatic β-cell destruction and is widely used to induce experimental diabetes. In this study; It was aimed to calculate the volume and cell numbers of the morphometric changes caused by streptozotocin-induced diabetes in the rat liver using stereological methods. Twenty adult male rats were used in the study. Four groups were created randomly. Blood glucose values of all groups were measured. Diabetes was induced by administering different doses of streptozotocin (55 mg/kg and 65 mg/kg). After 48 hours, blood glucose values were measured again. Four weeks later, liver tissues of all rats from the control and experimental groups were taken under anesthesia and fixed in 10% formaldehyde. The fixed liver tissues were divided into equal parts by physical fractionation. It was fixed in paraffin after passing through dehydration stages. Histological sections were prepared. It was stained with Hematoxylin-Eosin, examined under a light microscope and photographed. The total volume of the liver was calculated using the Cavalieri Principle and the number of hepatocytes was calculated with the unbiased counting frame of the dissector method. In the diabetes-related groups, the increase in liver volume and cell number was significant compared to the control group (p<0.05). As a result, streptozotocin injection and the resulting diabetes lead to an increase in liver volume and hepatocyte numbers.

Anahtar Kelimeler

Diabetes, Liver, Rat, Streptozotocin

Farklı Dozlarda Uygulanan Streptozotosinin Sıçan Karaciğerine Etkilerinin Stereolojik Metotlarla İncelenmesi

Öz

Diabetes Mellitus, vücudun insülin üretememesinden veya insülin etkisine karşı dirençten veya her ikisinden kaynaklanan yüksek kan şekeri seviyeleri ile karakterize edilen bir grup kronik metabolik rahatsızlıktır. İnsülin, vücut dokuları ve organları tarafından glikoz üretimini ve kullanımını düzenleyen ana hormondur. Diyabet, karaciğer de dahil olmak üzere vücuttaki tüm sistemleri etkiler. Streptozotosin, pankreas β-hücre yıkımı gerçekleştiren bir antibiyotiktir ve deneysel diyabet oluşturmak için yaygın olarak kullanılır. Bu çalışmada, sıçan karaciğerinde sterptozotosin kaynaklı diyabetin neden olduğu morfometrik değişikliklerin hacim ve hücre sayılarının stereolojik yöntemler kullanılarak hesaplanması amaçlandı. Çalışmada erişkin 20 adet erkek sıçan kullanıldı. Rastgele dört grup oluşturuldu. Tüm grupların kan glikoz değerleri ölçüldü. Streptozotosinin farklı dozları (55 mg/kg ve 65 mg/kg) verilerek diyabet indüklendi. 48 saat sonra tekrar kan glikoz değerleri ölçüldü. Dört hafta sonra kontrol ve denek gruplarına ait tüm sıçanların anestezi altında karaciğer dokuları çıkarılarak %10’luk formaldehitte tespit edildi. Tespit edilen karaciğer dokuları fiziksel fraksiyonlama ile eşit parçalara ayrıldı. Dehidratasyon aşamalarından geçerek parafinde sabitlendi. Histolojik kesitler hazırlandı. Hematoksilen-Eozin ile boyandı ve ışık mikroskobunda incelenerek fotoğraflandı. Cavalieri Prensibi ile karaciğerin total hacmi, disektör yönteminin tarafsız sayım çerçevesi ile hepatosit sayıları hesaplandı. Diyabete bağlı gruplarda, kontrol grubuna göre karaciğer hacminde ve hücre sayısındaki artış anlamlıydı (p<0.05). Sonuç olarak, streptozotosin enjeksiyonu ve bunun sonucunda ortaya çıkan diyabet, karaciğer hacminde ve hepatosit sayılarında artışa yol açmaktadır.

Anahtar Kelimeler

Diyabet, Karaciğer, Sıçan, Streptozotosin

Kaynakça

• Al-Ani IMD, Al-Mishadani NMS, Muslih RK, Hamoodi SR (2009). Histological liver changes in streptozotocin induced diabetic mice. Med J Malaysia, 8 (1).
• Alberti KGMM (2010). The classification and diagnosis of diabetes mellitus. Holt R, Cockram C, Flyvbjerg A and Goldstein B (Ed). Endocrinology and Metabolism (pp. 24-30). Blackwell Publishing, London, UK.
• Alshathly MR (2019). Efficacy of Ginger (Zingiber officinale) in ameliorating streptozotocin-induced diabetic liver injury in rats: Histological and biochemical studies. J Microsc Ultrastruct, 7 (2), 91-101.
• Balasubramanian T, Senthilkumar GP, Karthikeyan M, Chatterjee TK (2013). Protective effect of ethyl acetate fraction of stereo spermum suaveolens agains thepatic oxidative stress in STZ diabeticrats. J Tradit Complement Med, 3 (3), 175-181.
• Bilal HM, Riaz F, Munir K, Saqib A, Sarwar MR (2016). Histological changes in the liver of diabetic rats: A review of pathogenesis of non alcoholic fatty liver disease in type 1 diabetes mellitus. Cogent Medicine, 3 (1), 1275415.
• Chandirasegaran G, Elanchezhiyan C, Ghosh K (2018). Effects of Berberine chloride on the liver of streptozotocin-induced diabetes in albino Wistar rats. Biomed Pharmacother, 99, 227-236.
• Correia-Santos AM, Suzuki A, Anjos JS et al. (2012). Induction of Type 2 Diabetes by low dose of streptozotocin and high-fatdiet-fed in wistarrats. Medicina (Ribeirao Preto), 45 (4), 436-444.
• Deeds MC, Anderson JM, Armstrong AS et al. (2011). Single dose streptozotocin-induced diabetes: considerations for study design in is let transplantation models. Lab Anim, 45 (3), 131-140.
• Furman, BL (2021). Streptozotocin‐induced diabetic models in mice and rats. Curr Protoc., 1 (4), e78.
• Gundersen HJG (1986). Stereology of arbitrary particles a review of un biased number and size estimators and the presentation of some new ones, in memory of william r. thompson. J Microsc, 143 (1), 3-45.
• Gundersen HJG, Jensen EB (1987). The efficiency of systematic sampling in stereology and itsprediction. J Microsc, 147 (3), 229-263.
• Halici Z, Bilen H, Albayrak F et al. (2009). Does telmisartan prevent hepatic fibrosis in rats with alloxan-induced diabetes? Eur J Pharmacol, 614 (1-3), 146-152.
• Kohl T, Gehrke N, Schad A et al. (2013). Diabetic liver injury from streptozotocin is regulated through the caspase-8 homolog CFLIP involving activation of JNK2 and intrahepatic immunocompetent cells. Cell Death Dis, 4 (7), e712.
• Lucchesi AN, Cassettari LL, Spadella CT (2015). Alloxan-induced diabetes causes morphological and ultrastructural changes in rat liver that resemble the natural history of chronic fatty liver disease in humans. J Diabetes Res, 2015, 494578.
• Magalhaes DA, Kume WT, Correia FS et al. (2019). High-fat diet and streptozotocin in the induction of type 2 diabetes mellitus: a new proposal. An Acad Bras Cienc, 91 (1), e20180314.
• Maitra A, Abbas AK (2005). The endocrine pancreas. Robbins and Cotran pathologic basis of disease, 7th edn. Elsevier Saunders, Philadelphia. 1190-1205.
• Mazonakis M, Damilakis J, Maris T, Prassopoulos P, Gourtsoyiannis N (2002). Comparison of two volumetric techniques for estimating liver volume using magnetic resonance imaging. J Magn Reson Imaging, 15 (5), 557-563.
• Mohamed J, Nafizah AN, Zariyantey AH, Budin S (2016). Mechanisms of diabetes-induced liver damage: the role of oxidative stres and inflammation. Sultan Qaboos Univ Med J, 16 (2), e132.
• Piya MK, Tahranı AA, Barnett AH (2010). Emerging Treatment Options for Type 2 Diabetes. Brit J Clinical Pharma, 70 (5), 631-644.
• Punithavathi VR, Anuthama R, Prince PSM (2008). Combined treatment with naringin and vitamin C amelio rates streptozotocin‐induced diabetes in male wistar rats. J Appl Toxicol, 28 (6), 806-813.
• Saeedi P, Petersohn I, Salpea P et al. (2019). Diabetes Atlas Committee. Global andregional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Resultsfromthe International Diabetes Federation Diabetes Atlas. Diabetes Res Clin Pract, 157, 107843.
• Sipal S, Halici Z, Kiki I et al. (2012). Comparative study of three angiotensin II type 1 receptor antagonists in preventing liver fibrosis in diabetic rats: stereology, histopathology, and electron microscopy. J Mol Histol, 43 (6), 723-735.
• Szkudelski T (2001). The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas. Physiol Res, 50 (6), 537-546.
• Unal D, Aksak S, Halici Z et al. (2011) Effects of diabetes mellitus on the rat liver during the postmenopausal period. J Mol Histol 42 (3), 273-287.
• Ünal B, Canan S, Aslan H, ve ark. (2002). Doku örneklerindeki objelerin sayılarının hesaplanmasında tarafsız stereolojik metodlar: Fiziksel disektör. T Klin Tıp Bilimleri, 22, 15-24.
• Wang-Fischer Y, Garyantes T (2018). Improving there liability and utility of streptozotocin-induced rat diabetic model. J Diabetes Res, 2018, 8054073, 14 pages.
• Yaman T, Doğan A (2016). Streptozotosin ile diyabet oluşturulan sıçanlarda meşe palamudu (quercus brantilindl.) ekstraktların karaciğer ve pankreası koruyucu etkileri. Dicle Üniv Vet Fak Derg, 1, 7-15.
• Zangeneh MM, Zangeneh A, Bahrami E et al. (2018). Evaluation of hematoprotective and hepatoprotective properties of aqueous extract of Ceterach officinarum DC against streptozotocin-induced hepatic injury in male mice. Comp Clin Pathol, 27 (6), 1427-1436.
• Zheng Y, El-kott AF, Shaldoum F et al. (2021). Alleviation of Diabetes-Induced Hepatotoxicity by Date Palm Hydroalcoholic Extract in Rat Model; A Biochemical, Immunohistochemical and Stereological Study. Int J Morphol, 39 (3), 876-885.