Identification of vimentin,pancytokeratin, smooth muscle actin, Е-katherin and antibodies TOCD-IO -markers of epithelial and mesenchymal transformation in chronic glomerulonephritis

AIM: to study factors of epithelial and mesenchymal transformation in glomerulonephritis. PATIENTS AND METHODS. The study included 8 patients with various forms of chronic glomerulonephritis. Mean age of patients was 36.8 (22 to 60) years. To estimate epithelial cell phenotype we used monoclonal antibodies (company DAKO, ready-to-use (RTU) to pancytokeratin (clone AE1/AE3), E-cadherin (clone NCH-38), CD-10 (clone 56C6), as markers of mesenchymal differentiation we used monoclonal antibodies to vimentin (clone V9) and alpha-smooth muscle actin (clone 1A4). Evaluation of proliferative activity of tubular epithelium was performed with antibodies to Ki67 (clone MIB-1). RESULTS. EMT markers analysis showed the following. In areas where there were no signs of interstitial fibrosis pancytokeratin AE1/AE3 expression was determined along the membrane and cytoplasm of epithelial collecting ducts of cortex and medulla. In the epithelium of convoluted tubules reaction was less pronounced. Positive reaction with antibodies to CD-10 was observed in the epithelium and glomerular mesangial cells, mainly on the apical surface of convoluted and direct tubules epithelium, but was absent in the epithelium of collecting ducts. Expression of E-cadherin was revealed only in the epithelium of cortical and medullary collecting ducts. Differentiation mesenchymal marker vimentin was positive in the cytoplasm of single cells in convoluted tubules. CONCLUSION. The study confirmed possibility of changing of the epithelial phenotype to mesenchymal cells of the nephron tubules and ducts in various forms of glomerulonephritis. These changes are associated with the development of interstitial fibrosis. Increasing of number of proliferating cells in the areas of EMT possibly due to activation of reparative processes.

CD-10, vimentin, E-cadherin, smooth muscle actin, CD-10, chronic glomerulonephritis

1. Галишон П., Гертиг А. Эпителиально-мезенхимальная трансформация как биомаркер почечного фиброза: готовы ли мы применить теоретические знания на практике? Нефрология 2013; 17 (4): 9-16.

2. Репин В.С., Сабурина И.Н. Обратимые эпителиомезенхимальные траснформации клеток в эмбриогенезе и постнатальном обновлении тканей. Клет. транспл. и тканевая инженерия 2006; 1 (3): 64-72.

3. Biamonti G., Catillo M., Pignataro D. et al. The alternative splicing side of cancer. Semin Cell Dev Biol 2014; (14): 44-49.

4. Schindeler A., Kolind M., Little D.G. Cellular transitions and tissue engineering. Cell Reprogram 2013; 15(2): 101-107.

5. Kriz W., Kaissling B., Le Hir M. Epithelial-mesenchymal transition (EMT) in kidney fibrosis: fact or fantasy? J Clin Invest 2011; 121(2): 468-474.

6. Pieczynski J., Margolis B. Protein complexes that control renal epithelial polarity. Am J Physiol Renal Physiol 2011; 300(3): 589-601.

7. Lee J.M., Dedhar S., Kalluri R., Thompson E. The epithelialmesenchymal transition: new insights in signaling, development, and disease. J of Cell Biology 2006; 172(7): 973-981.

8. Lee K., Nelson C. New Insights into the Regulation of Epithelial-Mesenchymal Transition and Tissue Fibrosis. Int Review of Cell and Mol Biol 2012; (294): 171-221.

9. Galichon P., Hertig A. Epithelial to mesenchymal transition as a biomarker in renal fibrosis: are we ready for the bedside? Fibrogenesis Tissue Repair 2011; (6): 4-11.

10. Коржевский Д.Э., Ленцман М.В., Гиляров А.В. и др. Индукция синтеза нестина в клетках головного мозга крысы под влиянием ишемического повреждения. Морфология 2007; 131 (1): 23-26.

11. Коржевский Д.Э., Кирик О.В. Белки промежуточных филаментовнестин и виментин в клетках почки крысы. Морфология 2008; 134 (6): 50-55.

12. Sahli S., Stump B., Welti T. et al. A New Class of Inhibitors for the Metalloprotease Neprilysin Based on a Central Imidazole Scaffold. Helvetica Chimica Acta 2005; 88 (4): 707-730.

13. Чеботарёва Н.В., Бобкова И.Н., Варшавский В.А. и др. Роль гладкомышечного а-актина в развитии фиброза почек у больных хроническим гломерулонефритом. Тер. арх. 2006; 78 (5): 17-21.

14. Humphreys B.D., Czerniak S., DiRocco D.P. et al. Repair of injured proximal tubule does not involve specialized progenitors. Proc of the Nat Acad of Sciences of the USA 2011; (108): 9226-9231

15. Scholzen T., Gerdes J. The Ki-67 protein: from the known and the unknown. J Cell Physiol 2000; 182 (3): 311-322.