人白蛋白对HK-2细胞凋亡的作用

Effects of Human Albumin on the Apoptosis of HK-2 Cells

目的:探讨人白蛋白对体外培养的HK-2细胞凋亡的作用。方法:本实验研究对象为HK-2细胞株,将培养的HK-2细胞与20g/L的人白蛋白共同孵育0、4、6、8小时后,用Hoechst33258染色检测细胞凋亡。不同浓度(0g/L、5 g/L、10 g/L、20g/L和30g/L)的人白蛋白与体外培养的HK-2分别共同孵育0、4、6、8小时后,流式细胞仪检测细胞凋亡。结果:Hoechst33258染色结果显示:培养基对照组未见明显细胞凋亡;20g/L白蛋白与HK-2细胞共同孵育4、6、8小时,与对照组比较,均可见HK-2细胞荧光强度增加,有着典型凋亡形态的细胞增多,且随着人白蛋白与HK-2细胞作用时间的延长,细胞凋亡的程度和数目也增多。流式细胞仪检测结果显示:与对照组比较,HK-2细胞的凋亡率随着人白蛋白与HK-2细胞作用的浓度和时间增加而显著性增高,细胞凋亡率在8h组为(9.15±0.15%),在30g/L组为(9.35±0.46%),均为最高。结论:人白蛋白以时间和剂量依赖方式诱导肾小管细胞凋亡,以30g/L作用浓度和8小时作用时间的人白蛋白诱导HK-2细胞凋亡的作用最显著。

Objective： To investigate the effects of human albumin on the apoptosis of renal tubular epithelial HK-2 cells in vitro. Methods： HK-2 cells were incubated with various concentrations（0g/L, 5g/L, 10 g/L, 20 g/L, 30g/L） of human albumin for Oh, 4h, 6h and 8h. The apoptosis of HK-2 cells were evaluated by Hoechst33258 fluorescence stained cytospins and flow cytometry. Results： No significant apoptosis was observed in the control group treated by normal culture medium, but the fluorescence intensity and number of typical apoptotic cells increased in the cells treated with 20g/L human albumin for 4, 6 and 8 hours, and they increased gradually when the time extended. The flow cytometry results showed that the rates of apoptosis in HK-2 cells rose significantly along with increasing concentrations of human albumin and prolonging of incubation time, which was the highest at concentration of 30g/L and for 8h （respectively 9.15 ± 0.15% and 9.35 ±0.46% ）. Conclusions： Human albumin induced the apoptosis of HK-2 cells in a dose and time-dependent manner. It got the most significant effect when it lasted for 8h at concentration of 30g/L.