Sıçanlarda Metformin Kullanımının Karaciğer Dokuları Üzerine Morfolojik Etkilerinin Işık ve Elektron Mikroskopisi ile Değerlendirilmesi
Abstract
Metformin, tip II diyabet için en sık reçete edilen oral anti-diyabetik ajandır. Genellikle güvenli bir ilaç olarak kabul edilir, ancak hepatotoksik etkisi de olabileceği bildirilmektedir. Literatürde metforminin karaciğer dokusundaki morfolojik ve ultrayapısal etkilerini değerlendiren çalışmalar, daha çok deneysel olarak oluşturulan bir karaciğer hasarını azaltıp azaltmadığı yönündedir. Ancak bu ilaç polikistik over sendromu gibi, herhangi bir karaciğer rahatsızlığı olmayan hastalarda da kullanılabilmektedir. Bu çalışmadaki amacımız herhangi bir karaciğer hastalığı yok iken metformin kullanımının karaciğerde herhangi bir değişime neden olup olmadığını ışık ve geçirimli elektron mikroskopisi yardımı (TEM) ile değerlendirmektir. Bu amaçla Wistar Albino erkek sıçanlar kontrol ve metformin grubu olarak iki gruba ayrılmıştır. Metformin grubuna üç hafta boyunca metformin (100mg / kg) uygulanmış ve kontrol grubuna eşit hacimde serum fizyolojik verilmiştir. Bir haftalık ilaçsız dönemden sonra sıçanlardan ketamin/ksilazin anestezisi altında karaciğer örnekleri alınmış ve ötenazi uygulanmıştır. Biyopsi sonrası karaciğer örnekleri ışık ve elektron mikroskobik incelemeler için rutin TEM takibine alınmışlardır. Örneklerden elde edilen yarı ince (700 nm) ve TEM için alınan tam ince (60 nm) kesitler önce ışık mikroskobu ile sonra da Hitachi HT 7800 TEM ile değerlendirilmiştir. Kontrol grubu örneklerinde düzenli hepatosit, çekirdek ve çekirdekçik içeren sağlıklı bulgular izlenirken, metformin uygulanan örneklerde bol miktarda koful oluşumu, çekirdek düzensizlik ve küçülmeleri, bol kan damarı ve hepatosit ayrılmaları gözlenmiştir. Çalışmamızın bulguları metforminin uzun süreli kullanımda hepatositler üzerinde olumsuz etkileri olabileceğini göstermektedir.
Thanks
Bu çalışmanın, Deney hayvanlarına ilaç uygulanması aşaması Balıkesir Üniversitesi Deney Hayvanları Üretim, Bakım, Uygulama ve Araştırma Merkezi’nde (BAUN-DEHAM), Işık ve Elektron mikroskobik incelemeleri Eskişehir Osmangazi Üniversitesi Araştırma Laboratuvarı Uygulama ve Araştırma Merkezi'nde (ARUM) yapılmıştır.
References
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- Doycheva I., Loomba R. Effect of metformin on ballooning degeneration in nonalcoholic steatohepatitis (NASH): When to use metformin in nonalcoholic fatty liver disease (NAFLD) Adv. Ther. 2014;31:30–43. doi: 10.1007/s12325-013-0084-6.
- Dumitrescu R, Mehedintu C, Briceag I, Purcărea VL, Hudita D. Metformin-clinical pharmacology in PCOs. J Med Life. 2015 Apr-Jun;8(2):187-92.
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- Gharbaran R 2014. La Metformina Induce Alteraciones Ultraestructurales en Hepatocitos de Ratas Espontáneamente Hipertensas. International Journal of Morphology, 32(3): 839-843.
- Gopal V, Mandal V, Tangjang S, Mandal SC. Serum Biochemical, Histopathology and SEM Analyses of the Effects of the Indian Traditional Herb Wattakaka Volubilis Leaf Extract on Wistar Male Rats. J Pharmacopuncture. 2014;17(1):13–19. doi:10.3831/KPI.2014.17.002
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- Rena G, Hardie DG, Pearson ER. The mechanisms of action of metformin. Diabetologia. 2017 Sep;60(9):1577-1585. doi: 10.1007/s00125-017-4342-z.
- Saeedi P, Petersohn I, Salpea P, Malanda B, Karuranga S, Unwin N, Colagiuri S, Guariguata L, Motala AA, Ogurtsova K, Shaw JE, Bright D, Williams R; IDF Diabetes Atlas Committee. Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas, 9(th) edition. Diabetes Res Clin Pract. 2019 Nov;157:107843. doi:10.1016/j.diabres.2019.107843.
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- Song YM, Lee YH, Kim JW, Ham DS, Kang ES, Cha BS, Lee HC, Lee BW 2015. Autophagy, 11(1), 46-59.
- Tang X, Li J, Xiang W, Cui Y, Xie B, Wang X, Xu Z, Gan, L 2016. Metformin increases hepatic leptin receptor and decreases steatosis in mice. Journal of Endocrinology, 230(2), 227-37.
- Uygun A, Kadayifci A, Isik AT, Ozgurtas T, Deveci S, Tuzun A, Gulsen K, Dagalp K 2004. Metformin in the treatment of patients with non‐alcoholic steatohepatitis. Alimentary pharmacology & therapeutics, 19(5): 537-544.
- Wang YW, He SJ, Feng X, et al. Metformin: a review of its potential indications. Drug Des Devel Ther. 2017;11:2421–2429. doi:10.2147/DDDT.S141675
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- Zhou J, Massey S, Story D, Li L. Metformin: An Old Drug with New Applications. Int J Mol Sci. 2018 Sep 21;19(10). pii: E2863. doi: 10.3390/ijms19102863.
Evaluation of morphological effects on liver tissues of metformin use in rats by light and electron microscopy
Abstract
Metformin is the most commonly prescribed oral anti-diabetic agent for type II diabetes. It is generally considered as a safe drug but it is reported that it may also have a hepatotoxic effect. Studies in the literature evaluating the morphological and ultrastructural effects of metformin on liver tissue are more likely include whether it reduces an experimentally created liver injury. However, this drug can also be used in patients without any liver disease, such as polycystic ovarian syndrome. Our aim in this study is to evaluate whether metformin use causes any change in the liver with the help of light and permeable electron microscopy (TEM) while there is no liver disease. For this purpose, Wistar Albino male rats were divided into two groups as control and metformin groups. Metformin (100mg/kg) was applied to the metformin group for three weeks and an equal volume of saline was given to the control group. Liver samples were taken from rats under ketamine/xylazine anesthesia after one week of the drug-free period and euthanasia was performed. After biopsy, liver samples were taken for routine TEM procedure for light and electron microscopic examinations. Semithin (700 nm) and ultrathin (60 nm) sections from samples were evaluated by firstly light microscope and then Hitachi HT 7800 TEM. While healthy findings including regular hepatocyte, nucleus and nucleolus were observed in the control group samples, abundant vacuol formation, nuclear irregularity and shrinkage, abundant blood vessel and hepatocyte separation were observed in metformin-treated samples. The findings of our study show that metformin may have adverse effects on hepatocytes in long-term use.
References
- Ahishali E, Demir K, Ahishali B, Akyuz F, Pinarbasi B, Poturoglu S, İbrisim D, Gulluoglu M, Ozdil S, Besisik F, Kaymakoglu, S 2010. Electron microscopic findings in non‐alcoholic fatty liver disease: Is there a difference between hepatosteatosis and steatohepatitis?. Journal of gastroenterology and hepatology, 25(3): 619-626.
- Al‐Hashem F, Al‐Humayed S, Amin SN, Kamar SS, Mansy SS, Hassan S, Abdel-Salam OL, Ellatif MA,Alfaifi M, Haidara MA, Al‐Ani, B 2019. Metformin inhibits mTOR–HIF‐1α axis and profibrogenic and inflammatory biomarkers in thioacetamide‐induced hepatic tissue alterations. Journal of cellular physiology, 234(6): 9328-9337.
- Brunmair B, et al. Thiazolidinediones, like metformin, inhibit respiratory complex I: a common mechanism contributing to their antidiabetic actions? Diabetes. 2004;53:1052–1059.
- Cone CJ, Bachyrycz AM, Murata GH. Hepatotoxicity associated with metformin therapy in treatment of type 2 diabetes mellitus with nonalcoholic fatty liver disease. Ann Pharmacother. 2010 Oct;44(10):1655-9. doi: 10.1345/aph.1P099.
- de Souza Teixeira AA, Souza CO, Biondo LA, Sanches Silveira L, Lima EA,Batatinha HA, Araujo AP, Alves MJ, Hirabara SM, Curi R, Neto JCR. Short-term treatment with metformin reduces hepatic lipid accumulation but induces liver inflammation in obese mice. Inflammopharmacology. 2018 Aug;26(4):1103-1115. doi:10.1007/s10787-018-0443-7.
- Doycheva I., Loomba R. Effect of metformin on ballooning degeneration in nonalcoholic steatohepatitis (NASH): When to use metformin in nonalcoholic fatty liver disease (NAFLD) Adv. Ther. 2014;31:30–43. doi: 10.1007/s12325-013-0084-6.
- Dumitrescu R, Mehedintu C, Briceag I, Purcărea VL, Hudita D. Metformin-clinical pharmacology in PCOs. J Med Life. 2015 Apr-Jun;8(2):187-92.
- Foretz M, Guigas B, Bertrand L, Pollak M, Viollet B. Metformin: from mechanisms of action to therapies. Cell Metab. 2014 Dec 2;20(6):953-66. doi:10.1016/j.cmet.2014.09.018.
- Gharbaran R 2014. La Metformina Induce Alteraciones Ultraestructurales en Hepatocitos de Ratas Espontáneamente Hipertensas. International Journal of Morphology, 32(3): 839-843.
- Gopal V, Mandal V, Tangjang S, Mandal SC. Serum Biochemical, Histopathology and SEM Analyses of the Effects of the Indian Traditional Herb Wattakaka Volubilis Leaf Extract on Wistar Male Rats. J Pharmacopuncture. 2014;17(1):13–19. doi:10.3831/KPI.2014.17.002
- Hashmi T. Probable hepatotoxicity associated with the use of metformin in type 2 diabetes. BMJ Case Rep. 2011 Sep 13;2011. pii: bcr0420114092. doi:10.1136/bcr.04.2011.4092.
- Iranshahy M, Rezaee R, Karimi G. Hepatoprotective activity of metformin: A new mission for an old drug? Eur J Pharmacol. 2019 May 5;850:1-7. doi:10.1016/j.ejphar.2019.02.004.
- Kutoh E. Possible metformin-induced hepatotoxicity. Am J Geriatr Pharmacother. 2005 Dec;3(4):270-3.
- Lashen H 2010. Role of metformin in the management of polycystic ovary syndrome. Therapeutic advances in endocrinology and metabolism, 1(3): 117-128.
- Li R., Chen L.Z., Zhao W., Zhao S.P., Huang X.S. Metformin ameliorates obesity-associated hypertriglyceridemia in mice partly through the apolipoprotein A5 pathway. Biochem. Biophys. Res. Commun. 2016;478:1173–1178. doi: 10.1016/j.bbrc.2016.08.087.
- Lin M.J., Dai W., Scott M.J., Li R., Zhang Y.Q., Yang Y., Chen L.Z., Huang X.S. Metformin improves nonalcoholic fatty liver disease in obese mice via down-regulation of apolipoprotein A5 as part of the AMPK/LXRα signaling pathway. Oncotarget. 2017;8:108802–108809.
- LiverTox: Clinical and Research Information on Drug-Induced Liver Injury [Internet]. Bethesda (MD): National Institute of Diabetes and Digestive and Kidney Diseases; 2012-. 2020 Jan 21. Available from http://www.ncbi.nlm.nih.gov/books/NBK548726/
- Madiraju AK, Erion DM, Rahimi Y, et al. Metformin suppresses gluconeogenesis by inhibiting mitochondrial glycerophosphate dehydrogenase. Nature. 2014;510:542–546. doi: 10.1038/nature13270.
- Martin-Montalvo A, Mercken EM, Mitchell SJ, Palacios HH, Mote PL, Scheibye-Knudsen M, Gomes AP, Ward TM, Minor RK, Blouin MJ, Schwab M, Pollak M, Zhang Y, Yu Y, Becker KG, Bohr VA, Ingram DK, Sinclair DA, Wolf NS, Spindler SR, Bernier M, de Cabo R. Metformin improves healthspan and lifespan in mice. Nat Commun. 2013;4:2192. doi: 10.1038/ncomms3192.
- Meng XM, Ma XX, Tian YL, Jiang Q, Wang LL, Shi R, Ding L, Pang SG 2017. Metformin improves the glucose and lipid metabolism via influencing the level of serum total bile acids in rats with streptozotocin-induced type 2 diabetes mellitus. European Review for Medical and Pharmacological Sciences, 21(9): 2232-2237.
- Rena G, Hardie DG, Pearson ER. The mechanisms of action of metformin. Diabetologia. 2017 Sep;60(9):1577-1585. doi: 10.1007/s00125-017-4342-z.
- Saeedi P, Petersohn I, Salpea P, Malanda B, Karuranga S, Unwin N, Colagiuri S, Guariguata L, Motala AA, Ogurtsova K, Shaw JE, Bright D, Williams R; IDF Diabetes Atlas Committee. Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas, 9(th) edition. Diabetes Res Clin Pract. 2019 Nov;157:107843. doi:10.1016/j.diabres.2019.107843.
- Soliman HM, Wagih HM, Algaidi SA, Hafiz AH 2013. Histological evaluation of the role of atypical antipsychotic drugs in inducing non-alcoholic fatty liver disease in adult male albino rats (light and electron microscopic study). Folia biologica, 59(5): 173.
- Song YM, Lee YH, Kim JW, Ham DS, Kang ES, Cha BS, Lee HC, Lee BW 2015. Autophagy, 11(1), 46-59.
- Tang X, Li J, Xiang W, Cui Y, Xie B, Wang X, Xu Z, Gan, L 2016. Metformin increases hepatic leptin receptor and decreases steatosis in mice. Journal of Endocrinology, 230(2), 227-37.
- Uygun A, Kadayifci A, Isik AT, Ozgurtas T, Deveci S, Tuzun A, Gulsen K, Dagalp K 2004. Metformin in the treatment of patients with non‐alcoholic steatohepatitis. Alimentary pharmacology & therapeutics, 19(5): 537-544.
- Wang YW, He SJ, Feng X, et al. Metformin: a review of its potential indications. Drug Des Devel Ther. 2017;11:2421–2429. doi:10.2147/DDDT.S141675
- Zheng L. Metformin as a Rare Cause of Drug-Induced Liver Injury, a Case Report and Literature Review. Am J Ther. 2016 Jan-Feb;23(1):e315-7. doi:10.1097/MJT.0000000000000007.
- Zhou J, Massey S, Story D, Li L. Metformin: An Old Drug with New Applications. Int J Mol Sci. 2018 Sep 21;19(10). pii: E2863. doi: 10.3390/ijms19102863.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Clinical Sciences |
| Journal Section | Research Articles |
| Authors | |
| Publication Date | June 22, 2020 |
| Acceptance Date | June 17, 2020 |
| Published in Issue | Year 2020 Volume: 3 Issue: 1 |
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