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作 者:张林 刘伏友[2] 彭佑铭[2] 段绍斌[2] 刘虹[2] 李瑛[2]
机构地区:[1]长沙市四医院肾内科,湖南长沙410006 [2]中南大学湘雅二医院肾脏病研究所,湖南长沙410011
出 处:《现代生物医学进展》2012年第26期5041-5044,共4页Progress in Modern Biomedicine
摘 要: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.
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