The effect of hyperosmolar stimuli and cyclophosphamide on the culture of normal rat urothelial cells in vitro

Cellular & Molecular Biology Letters

Paulina Chorobik²  Piotr Jan Thor³  Agata Ziomber³  Jolanta Kaszuba-Zwoińska³  Kajetan Juszczak¹ 

[1] Department of Pathophysiology, Jagiellonian University, Medical College, Cracow, Poland$$Department of Urology, Rydygier Memorial Hospital, Cracow, Poland$$;Department of Immunology, Jagiellonian University, Medical College, Cracow, Poland$$;Department of Pathophysiology, Jagiellonian University, Medical College, Cracow, Poland$$

关键词: Urothelial cell;    Culture;    Cyclophosphamide;    Hyperosmolarity;    Bladder;    Rat;    Apoptosis;    Necrosis;    Overactive bladder;   

DOI  :  10.2478/s11658-012-0002-y

学科分类：分子生物学，细胞生物学和基因

来源: Uniwersytet Wroclawski * Wydzial Biotechnologii / University of Wroclaw, Faculty of Biotechnology

【 摘 要 】

Highly concentrated urine may induce a harmful effect on the urinary bladder. Therefore, we considered osmolarity of the urine as a basic pathomechanism of mucosal damage. The influence of both cyclophosphamide (CYP) and hyperosmolar stimuli (HS) on the urothelium are not well described. The purpose was to evaluate the effect of CYP and HS on rat urothelial cultured cells (RUCC). 15 Wistar rats were used for RUCC preparation. RUCC were exposed to HS (2080 and 3222 mOsm/l NaCl) for 15 min and CYP (1 mg/ml) for 4 hrs. APC-labelled annexin V was used to quantitatively determine the percentage of apoptotic cells and propidium iodide (PI) as a standard flow cytometric viability probe to distinguish necrotic cells from viable ones. Annexin V-APC (+), annexin V-APC and PI (+), and PI (+) cells were analysed as apoptotic, dead, and necrotic cells, respectively. The results were presented in percentage values. The flow cytometric analysis was done on a FACSCalibur Flow Cytometer using Cell-Quest software. Treatment with 2080 and 3222 mOsm/l HS resulted in 23.7 ± 3.9% and 26.0 ± 1.5% apoptotic cells, respectively, 14.3 ± 1.4% and 19.4 ± 2.7% necrotic cells, respectively and 60.5 ± 1.4% and 48.6 ± 5.3% dead cells, respectively. The effect of CYP on RUCC was similar to the effect of HS. After CYP the apoptotic and necrotic cells were 23.1 ± 0.3% and 17.9 ± 7.4%, respectively. The percentage of dead cells was 57.7 ± 10.8%. CYP and HS induced apoptosis and necrosis in RUCC. 3222 mOsm/l HS had the most harmful effect based on the percentage of necrotic and apoptotic cells.

【 授权许可】

Unknown