de Souza Durão Marcelino, Razvickas Clara Versolato, Gonçalves Elsa Alídia Petry, Okano Iria Ruriko, Camargo Simone Mafalda Rodrigues, Monte Júlio Cesar Martins, dos Santos Oscar Fernando Pavão
Nephrology Division, Universidade Federal de São Paulo, Escola Paulista de Medicina, São Paulo, Brazil.
Ren Fail. 2003 May;25(3):341-53. doi: 10.1081/jdi-120021149.
In acute renal failure (ARF) renal tubular cell death and detachment can be induced by necrotic and apoptotic mechanisms. Several studies have demonstrated some benefits of the use of growth factors in experimental models of ARF.
MDCK cells were cultured in a glucose-free medium for 24h and were submitted to hypoxia (PO2 around 35 mmHg) for additional 24 h. To evaluate the possible protective role of growth factors, EGF, IGF-I or HGF were added to the medium (20 ng mL). LDH release, viability (acridine orange and ethidium bromide dyes) and quantification of apoptotic cells (Hoechst 33342 dye fluorescence) were determined.
In the injury group, an increase on LDH release (60% vs. 3%) and on number of apoptotic cells (22% vs. 0.2%) which was associated with a reduced cell viability (61% vs. 94%) when compared with controls. Only HGF, not EGF or IGF-I, was able to protect cells from injury. HGF caused a significant reduction on LDH release (30%) and on number of apoptotic cells (5%), with an increase on viability cellular (79%).
HGF decreases cell death on MDCK cells after hypoxic-induced injury, probably acting in both necrotic and apoptotic mechanisms.
在急性肾衰竭(ARF)中,肾小管细胞死亡和脱落可由坏死和凋亡机制诱导。多项研究已证明在ARF实验模型中使用生长因子有一些益处。
将MDCK细胞在无葡萄糖培养基中培养24小时,然后再进行24小时的缺氧处理(PO2约为35 mmHg)。为评估生长因子的可能保护作用,将表皮生长因子(EGF)、胰岛素样生长因子-I(IGF-I)或肝细胞生长因子(HGF)添加到培养基中(20 ng/mL)。测定乳酸脱氢酶(LDH)释放、细胞活力(吖啶橙和溴化乙锭染料)以及凋亡细胞定量(Hoechst 33342染料荧光)。
与对照组相比,损伤组中LDH释放增加(60%对3%),凋亡细胞数量增加(22%对0.2%),同时细胞活力降低(61%对94%)。只有HGF能够保护细胞免受损伤,而EGF或IGF-I则不能。HGF使LDH释放显著减少(30%),凋亡细胞数量显著减少(5%),细胞活力增加(79%)。
HGF可减少缺氧诱导损伤后MDCK细胞的死亡,可能通过坏死和凋亡机制发挥作用。