Zeynep Deniz ŞAHİN İNAN3, Serpil Ünver SARAYDIN

Evaluation of VEGF, Cytokeratin-19 and Caspase 3 Immunolocalization in the Lung Tissue of Rat with Experimentally Induced Diabetes

Diabetes Mellitus (DM) manifests itself with changes in the functional structure of the lungs and impairments in gas exchange. These changes in diabetic lung tissue may be due to various factors. Our aim in this study is to correlate the damage of diabetes with lung tissue in terms of VEGF, CK19, caspase 3 immunolocalizations. In this study, animals were divided into 4 groups, 60 mg/kg streptozotocin was given to each of the groups with experimental diabetes and the physiological saline solution was given intraperitoneally to the control group. On days 7 and 14 of the experiment, diabetic and control groups were euthanized, and lung tissues were removed. Tissue samples were evaluated histochemically and immunohistochemically by monitoring with standard light microscopy. In the diabetic group, the localization of CK19 and Caspase 3 increased on the 7th and 14th days compared to the control group, but the immunolocalization of VEGF decreased. Based on our findings, it was determined that lung tissue was one of the target organs of diabetes. The increase in pulmonary parenchyma due to hyperglycemia is accepted as a source of fibrosis. We concluded that due to increased CK19 localization of fibrosis source, decreased VEGF localization has increased apoptosis in the pulmonary capillary endothelium, which has a significant role in the blood-air barrier in the lung parenchyma, especially in endothelial cells.

Deneysel Olarak Diyabet Oluşturulan Ratların Akciğer Dokusunun Caspase 3, Cytokeratin 19 ve VEGF İmmünolokalizasyonunun Değerlendirilmesi

Diabetes Mellitus (DM), akciğerlerin fonksiyonel yapısındaki değişikliklerle ve gaz değişimlerindeki bozukluklarla kendini gösterir. Diyabetik akciğer dokusunda bu değişiklikler çeşitli faktörlere bağlı olabilir. Bu çalışmada amacımız VEGF, CK19, kaspaz 3 immunolokalizasyonları ile ilgili olarak diyabetin akciğer dokusuna verdiği zararı ilişkilendirmektir. Çalışmada hayvanlar 4 gruba ayrıldı, deneysel diyabetli gruplara 60 mg/kg streptozotosin verildi ve kontrol grubuna serum fizyolojik solüsyonu intraperitoneal olarak verildi. Deneyin 7 ve 14. günlerinde diyabetik ve kontrol grupları sakrifiye edildi ve akciğer dokuları çıkarıldı. Doku örnekleri, standart ışık mikroskobu ile izlenerek histokimyasal ve immünhistokimyasal olarak değerlendirildi. Diyabetik grupta 7. ve 14. günlerde kontrol grubu ile kıyaslandığında CK19 ve Kaspaz 3 lokalizasyonu artmış, ancak VEGF immunolokalizasyonu azalmıştır. Bulgularımıza göre, akciğer dokusu diyabetin hedef organlarından biri olduğunu göstermektedir. Hiperglisemiye bağlı pulmoner parankimde artış fibrozis kaynağı olarak kabul edildi. Sonuç olarak, pulmoner fibrozis kaynağının CK19 immünolokalizasyonunun artmasına bağlı olarak, azalmış VEGF lokalizasyonunun, özellikle endotelyal hücrelerde, akciğer parankimindeki kan-hava bariyerinde önemli bir rolü olan, pulmoner kapiler endotelyumda apoptosisi arttırdığı sonucuna varıldı.

PDF

___

1. Bahadır Erdoğan B, Uzaslan E: Diabet ve akciğer. Uludağ Üniv Tıp Fak Derg, 31 (1): 71-74, 2005.

2. Liao YF, Yin S, Chen ZQ, Li F, Zhao B: High glucose promotes tumor cell proliferation and migration in lung adenocarcinoma via the RAGENOXs pathway. Mol Med Rep, 17 (6): 8536-8541, 2018. DOI: 10.3892/ mmr.2018.8914

3. Nicolaie T, Zavoianu C, Nuta P: Pulmonary involvement in diabetes mellitus. Rom J Intern Med, 41, 365-374, 2003.

4. Hsia CCW, Raskin P: The diabetic lung: Relevance of alveolar microangiopathy for the use of inhaled insulin. Am J Med, 118, 205-211, 2005. DOI: 10.1016/j.amjmed.2004.09.019

5. Masiello P, Broca C, Gross R, Roye M, Manteghetti M, HillaireBuys D, Novelli M, Ribes G: Experimental NIDDM: Development of a new model in adult rats administered streptozotocin and nicotinamide. Diabetes, 47, 224-229, 1998. DOI: 10.2337/diab.47.2.224

6. Szkudelski T: The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas. Physiol Res, 50, 537-546, 2001.

7. Kehler DS, Stammers AN, Susser SE, Hamm NC, Kimber DE, Hlynsky MW, Duhamel TA: Cardiovascular complications of type 2 diabetes in youth. Biochem Cell Biol, 93 (5): 496-510, 2015. DOI: 10.1139/bcb-2014- 0118

8. Neufeld G, Cohen T, Gengrinovitch S, Poltorak Z: Vascular endothelial growth factor (VEGF) and its receptors. FASEB J, 13, 9-22, 1999.

9. Kolluru GK, Bir SC, Kevil CG: Endothelial dysfunction and diabetes: Effcts on angiogenesis, vascular remodeling, and wound healing. Int J Vasc Med, 2012:918267, 2012. DOI: 10.1155/2012/918267

10. Paneni F, Bekman JA, Creager MA, Cosentino F: Diabetes and vascular disease: Pathophysiology, clinical consequences, and medical therapy: Part I. Eur Heart J, 34 (31): 2436-2443, 2013. DOI: 10.1093/ eurheartj/eht149

11. İrtegün S, Deveci E: Diyabetik sıçanların testis dokusunda VEGF ve Bcl-2 ekspresyon düzeylerinin immünohistokimya ve western blot yöntemleri ile incelenmesi. Dicle Med J, 43 (4): 527-533, 2016.

12. Adler A: Obesity and target organ damage: Diabetes. Int J Obes, 26, 11-14, 2002. DOI: 10.1038/sj.ijo.0802212

13. Yilmaz BO, Yildizbayrak N, Aydin Y, Erkan M: Evidence of acrylamide- and glycidamide-induced oxidative stress and apoptosis in Leydig and Sertoli cells. Hum Exp Toxicol, 36 (12): 1225-1235, 2017. DOI: 10.1177/0960327116686818

14. Dobashi N, Fujita J, Ohtsuki Y, Yamadori I, Yoshinouchi T, Kamei T, Takahara J: Elevated serum and BAL cytokeratin 19 fragment in pulmonary fibrosis and acute interstitial pneumonia. Eur Respir J, 14 (3): 574-578, 1999. DOI: 10.1034/j.1399-3003.1999.14c15.x

15. Fujita J, Dobashi N, Ohtsuki Y, Yamadori I, Yoshinouchi T, Kamei T, Tokuda M, Hojo S, Okada H, Takahara J: Elevation of anti-cytokeratin 19 antibody in sera of the patients with idiopathic pulmonary fibrosis and pulmonary fibrosis associated with collagen vascular disorders. Lung, 177 (5): 311-319, 1999.

16. Iyonaga K, Miyafuji M, Suga M, Saita N, Ando M: Alterations in cytokeratin expression by the alveolar lining epithelial cells in lung tissue from patients with idiopathic pulmonary fibrosis. J Pathol, 182, 217-224, 1997. DOI: 10.1002/(SICI)1096-9896(199706)182:2<217::AIDPATH833>3.0.CO;2-A

17. Nakamura H, Abe S, Shibata Y, Yuki H, Suzuki H, Saito H, Sata M, Kato S, Tomoike H: Elevated levels of cytokeratin 19 in the bronchoalveolar lavage flid iof patients with chronic airway inflmmatory disease: A specific marker for bronchial epithelial injury. Am J Respir Crit Care Med, 155, 1217-1221, 1997. DOI: 10.1164/ajrccm.155.4.9105057

18. Iraz M, Ozerol E, Gulec M, Tasdemir S, Idiz N, Fadıllıoğlu E, Naziroglu M, Akyol O: Protective effct of caffic acid phenethyl ester (CAPE) administration on cisplatin-induced oxidative damage to liver in rat. Cell Biochem Funct, 24, 357-361, 2006. DOI: 10.1002/cbf.1232

19. Oliveira TL, Candeia-Medeiros N, Cavalcante-Araújo PM, Melo IS, Fávaro-Pípi E, Fátima LA, Rocha AA, Goulart LR, Machado UF, Campos RR, Sabino-Silva R: SGLT1 activity in lung alveolar cells of diabetic rats modulates airway surface liquid glucose concentration and bacterial proliferation. Sci Rep, 6:21752, 2016. DOI: 10.1038/srep21752

20. Aktaş MS, Kandemir FM, Özkaraca M, Hanedan B, Kırbaş A: Protective effcts of rutin on acute lung injury induced by oleic acid in rats. Kafkas Univ Vet Fak Derg, 23 (3): 443-451, 2017. DOI: 10.9775/ kvfd.2016.16977

21. Sime PJ, Marr RA, Gauldie D, Xing Z, Hewlett BR, Graham FL, Gauldie J: Transfer of tumor necrosis factor-α to rat lung induces severe pulmonary inflammation and patchy interstitial fibrogenesis withinduction of transforming growth factor-β1 and myofibroblasts. Am J Pathol, 153 (3): 825-832, 1998. DOI: 10.1016/S0002-9440(10)65624-6

22. Huang TW: Chemical and histochemical studies of human alveolar collagen fibers. Am J Pathol, 86 (1): 81-98, 1977.

23. Zhou X, Moore BB: Lung section staining and microscopy. Bio Protoc, 7 (10):e2286, 2017. DOI: 10.21769/BioProtoc.2286

24. Mo A: Pulmonary complications in diabetes mellitus. Mymensingh Med J, 23 (3): 603-605, 2014.

25. Marvisi M, Marani G, Brianti M, Della Porta R: Pulmonary complications in diabetes mellitus. Recenti Prog Med, 87 (12): 623-627, 1996.

26. Guo S, Meng XW, Yang XS, Liu XF, Ou-Yang CH, Liu C: Curcumin administration suppresses collagen synthesis in the hearts of rats with experimental diabetes. Acta Pharmacol Sin, 39, 195-204, 2018. DOI: 10.1038/aps.2017.92

27. Waisundara VY, Hsu A, Huang D, Tan BKH: Scutellaria baicalensis: Enhances the anti-diabetic activity of metformin in streptozotocininduced diabetic Wistar rats. Am J Chinese Med, 36, 517-540, 2008. DOI: 10.1142/S0192415X08005953

28. Komoike Y, Matsuoka M: Endoplasmic reticulum stress-mediated neuronal apoptosis by acrylamide exposure. Toxicol Appl Pharmacol, 310, 68-77, 2016. DOI: 10.1016/j.taap.2016.09.005

29. Dalquen P: The lung in diabetes mellitus. Respiration, 66, 12-13, 1999. DOI: 10.1159/000029330

30. Braun L, Kardon T, Reisz-Porszasz Z, Banhegyi G, Mandl J: The regulation of the induction of vascular endothelial growth factor at the onset of diabetes in spontaneously diabetic rats. Life Sci, 69 (21): 2533- 2542, 2001. DOI: 10.1016/s0024-3205(01)01327-3

31. Sisman AR, Kiray M, Camsari UM, Evren M, Ateş M, Baykara B, Aksu I, Güvendi G, Uysal N: Potential novel biomarkers for diabetic testicular damage in streptozotocin induced diabetic rats: Nerve growth factor beta and vascular endothelial growth factor. Dis Markers, 2014:108106, 2014. DOI: 10.1155/2014/108106