İsmail SEÇKİN, Mümin UZUNALAN, Meltem AYAZ PEKYAK, Sibel KÖKTÜRK, Hüseyin Avni SÖNMEZ, Zeynep Banu GÜNGÖR, Elif Yaprak SARAÇ, Aslı EMİNCİK, Sibel DEMİRCİ DELİPINAR

Deneysel Kronik Nefroz Modeli Sıçan Glomerüllerinde TGF-b1 Ekspresyonu ve Mezangiyal Değişiklikler

Amaç: Puromisin aminonükleozid nefroz (PAN), proteinüri ile seyreden podosit yaralanması modeli olarak yaygın bir şekilde kullanılmaktadır. Deneysel olarak uyarılmış PAN hayvan modellerinde, minimal değişikliklere sahip insan nefrotik sendromu ve fokal segmental glomerulosklerozdakine benzer histolojik değişiklikler gösterilmiş, ancak etki mekanizması tam olarak anlaşılamamıştır. Yapılan çalışmalar, çoğu kronik böbrek hasarlarında, ekstrasellüler matriks (ESM) birikiminin görüldüğü ve bu süreçte transforming büyüme faktörü-β1 (TGF-β1)’in anahtar bir rol oynadığı gösterilmiştir. Bu çalışmada biz, kronik PAN uyarılı sıçan böbrek glomerüllerinde, TGF-β1 ekspresyonu ve mezangial matriks (MM) birikimi arasındaki ilişkiyi, böbrek fonksiyonu ve ultrastrüktürel değişikliklerle incelemeyi amaçladık. Yöntemler: Bu çalışma için 12 erkek Wistar albino sıçan iki gruba ayrıldı: kontrol grubu ve kronik grup (n=6). Bütün veriler istatistiksel olarak One-way ANOVA testi ile analiz edildi. Bulgular: Proteinüri seviyeleri kronik nefroz grubunda kontrolden yüksek (p<0,0025), serum albümin ve kreatinin klirens değerleri ise oldukça düşüktü (p<0,05). PAN nefrozlu sıçanların glomerüllerinde, hem TGF-β1 ekspresyonu hem de MM birikimi belirgin şekilde artmıştı. Kronik PAN grubunun glomerüllerinin ultrastrüktürel görüntülerinde artmış MM içerisinde apoptotik endotel ve mezangial hücrelere, interstisyal kollajene ve makrofajlara rastladık. Sonuç: Bulgularımız, kronik PAN uygulamasının böbrek fonksiyonlarında bozulma ile birlikte, glomerülde TGF-β1 ekspresyonu, MM artışına, interstisyal kollajen oluşumuna ve makrofajların göçüne neden olan bir nefrotik sendromu indüklediğini düşündürmektedir. Bu çalışma, ikili işaretlemelerle TGF-β1’in ana kaynağının ve TGF-β1 sinyal yolaklarının belirlenmesi için daha ileri çalışmalara ihtiyaç duymaktadır.Cite this article as: Seçkin İ, Uzunalan M, Ayaz Pekpak M, Köktürk S, Sönmez HA, Güngör ZB. TGF-β1 expression and mesangial alterations in rat glomeruli of an experimental chronic nephrosis model. Cerrahpasa Med J 2019; 43(1): 6-12.

Anahtar Kelimeler:

Mezangial matriks, puromisin aminonükleozid nefroz, TGF-β1, ultrastrüktür

TGF-β1 expression and mesangial alterations in rat glomeruli of an experimental chronic nephrosis model

Objective: Puromycine aminonucleoside nephrosis (PAN) is extensively used as an experimental model of proteinuria following podocyte injury. In PAN animal models, nephrotic syndrome with minimal change disease and focal segmental sclerosis-like nephritis is demonstrated to be similar to that in human; however, the real mechanism of PAN is not yet elucidated. Evidence shows that transforming growth factor-β1 (TGF-β1) plays a key via stimulating matrix protein synthesis of both glomerular epithelial and mesangial cells. In this study, we aimed to examine the relationship between TGF-β expression and accumulation in mesangial matrix (MM) by the changes in renal function and ultrastructure alterations in renal glomeruli of the chronic PAN induced rats.Methods: Twelve male Wistar albino rats were divided into two groups: control group (n=6), chronic group (n=6). All data was statistically analyzed by One-way ANOVA Test. Results: Proteinuria levels in the chronic nephrosis groups were greater than the control (p<0.0025) whereas the levels of serum albumin and creatinine clearances were progressively decreased (p<0.05). TGF-β1 expression and MM were both significantly increased in PAN group. Ultrastructural images of glomeruli of PAN showed apoptotic endothelium and mesangial cells, interstitial collagen and macrophages in increased MM. Conclusion: Our findings suggest that chronic PAN application induces a nephrotic syndrome, leading to renal impairment by TGF-β1 expression in glomeruli, increase in MM, formation of abundant interstitial collagen, and migration of macrophages. We suggest that chronic PAN progressively increases TGF-β1 expression and apoptosis following expansion of mesangial matrix in the affected glomeruli. This study needs further researches to determine the main source and signal pathway of TGF-β1 expression by double staining methods.Cite this article as: Seçkin İ, Uzunalan M, Ayaz Pekpak M, Köktürk S, Sönmez HA, Güngör ZB. TGF-β1 expression and mesangial alterations in rat glomeruli of an experimental chronic nephrosis model. Cerrahpasa Med J 2019; 43(1): 6-12.

Keywords:

Mesangial matrix, puromycine aminonucleoside nephritis, TGF-β1, ultrastructure,

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1. Krishnamurti U, Zhou B, Fan WW, Tsilibary E, Wayner E, Kim Y, et al. Puromycin aminonucleoside suppresses integrin expression in cultured glomerular epithelial cells. J Am Soc Nephrol 2001; 12: 758-66. 2. Sanwal V, Pandya M, Bhaskaran M, Franki N, Reddy K, et al. Puromycin aminonucleoside induces glomerular epithelial cell apoptosis. Exp Mol Pathol 2001; 70: 54-64. 3. Savill J, Mooney A, Hughes J. Apoptosis and renal scarring. Kidney Int Suppl 1996; 54: S14-7. 4. Makino H, Shikata K, Wieslander J, Wada J, Kashihara N, Yoshioka K, et al. Localization of fibril/microfibril and basement membrane collagens in diabetic glomerulosclerosis in type 2 diabetes. Diabet Med 1994; 11: 304-11. 5. Marinides GN, Groggel GC, Cohen AH, Border WA. Enalapril and low protein reverse chronic puromycin aminonucleoside nephropathy. Kidney Int 1990; 37: 749-57. 6. Raij L, Azar S, Keane W. Mesangial immune injury, hypertension, and progressive glomerular damage in Dahl rats. Kidney Int 1984; 26: 137-43. 7. Hill C, Flyvbjerg A, Grønbaek H, Petrik J, Hill DJ, Thomas CR, et al. The renal expression of transforming growth factor-β isoforms and their receptors in acute and chronic experimental diabetes in rats. Endocrinology 2000; 141: 1196-208. 8. Kihara I, Yaoita E, ,Yamamoto T. Cellular processes of glomerular adhesion in aged rats. Acta Medica et Biologica 1990; 38: 69-80. 9. Shimizu A, Kitamura H, Masuda Y, Ishizaki M, Sugisaki Y, Yamanaka N. Apoptosis in the repair process of experimental proliferative glomerulonephritis. Kidney Int 1995; 47: 114-21. 10. Morioka Y, Koike H, Ikezumi Y, Ito Y, Oyanagi A, Gejyo F, et al. Podocyte injuries exacerbate mesangial proliferative glomerulonephritis. Kidney Int 2001; 60: 2192-204. 11. Kaplan L. Pesce Al Clinical Chemistry Theory, Analysis and Correlation. St. Louis, Mo: CV Mosby Co, 1984. 1231. 12. Seckin I, Uzunalan M, Pekpak M, Kokturk S, Sonmez H, Oztürk Z, et al. Experimentally induced puromycine aminonucleoside nephrosis (PAN) in rats: evaluation of angiogenic protein platelet-derived endothelial cell growth factor (PD-ECGF) expression in glomeruli. J Biomed Sci 2012; 19: 24. 13. Powell H. Relationship between proteinuria and epithelial cell changes in minimal lesion glomerulopathy. Nephron 1976; 16: 310-7. 14. Davies M, Shewring L, Thomas G, Jenner L. Stimulation of proteoglycan (pg) synthesis in rat mesangial cells (rmc) in response to tumor necrosing factor (tnf). In kidney international 1989. 15. Okuda S, Languino LR, Ruoslahti E, Border WA. Elevated expression of transforming growth factor-beta and proteoglycan production in experimental glomerulonephritis. Possible role in expansion of the mesangial extracellular matrix. J Clin Invest 1990; 86: 453-62. 16. Border WA, Noble NA. Transforming growth factor β in tissue fibrosis. N Engl J Med 1994; 331: 1286-92. 17. Lafayette RA, Mayer G, Park SK, Meyer TW. Angiotensin II receptor blockade limits glomerular injury in rats with reduced renal mass. J Clin Invest 1992; 90: 766-71. 18. Pawluczyk IZ, Harris KP. Harris, Transforming growth factor-β suppresses macrophage-induced mesangial cell fibronectin expression. Kidney Int 2001; 60: 533-42. 19. Sharma R1, Khanna A, Sharma M, Savin VJ. Transforming growth factor-β1 increases albumin permeability of isolated rat glomeruli via hydroxyl radicals. Kidney Int 2000; 58: 131-6. 20. Akagi Y, Isaka Y, Arai M, Kaneko T, Takenaka M, Moriyama T, et al. Inhibition of TGF-β1 expression by antisense oligonucleotides suppressed extracellular matrix accumulation in experimental glomerulonephritis. Kidney Int 1996; 50: 148-55. 21. Lee HS, Song CY. Differential role of mesangial cells and podocytes in TGF-beta-induced mesangial matrix synthesis in chronic glomerular disease. Histol Histopathol 2009; 24: 901-8. 22. Sandau K, Pfeilschifter J, Brüne B. Nitric oxide and superoxide induced p53 and Bax accumulation during mesangial cell apoptosis. Kidney Int 1997; 52: 378-86. 23. Johnson R, Yamabe H, Chen YP, Campbell C, Gordon K, Baker P, et al. Glomerular epithelial cells secrete a glomerular basement membrane-degrading metalloproteinase. J Am Soc Nephrol 1992; 2: 1388-97. 24. Johnson RJ, Lovett D. In vivo gene transfer, Koch’s postulates, and renal disease. J Clin Invest 1993; 92: 2568. 25. Shimizu A, Masuda Y, Kitamura H, Ishizaki M, Sugisaki Y, Yamanaka N. Apoptosis in progressive crescentic glomerulonephritis. Lab Invest 1996; 74: 941-51.