鱼胆汁毒素致肾小管上皮细胞死亡方式的试验研究

The experimental study on the death way of human renal epithelial cells treated by ichthyogalltoxin

目的观察草鱼胆汁及鲤醇硫酸酯钠致人肾近曲小管上皮细胞(HK-2)的死亡方式。方法根据文献报道方法从草鱼胆汁中提取鲤醇硫酸酯钠,通过体外培养HK-2细胞系,应用MTT法评价草鱼胆汁和鲤醇硫酸酯钠对HK-2细胞增殖抑制作用,应用流式细胞术检测草鱼胆汁和鲤醇硫酸酯钠所致的HK-2细胞的主要死亡方式。结果 HK-2细胞存活率随着草鱼胆汁和鲤醇硫酸酯钠浓度的增加和时间的延长而降低,且草鱼胆汁和鲤醇硫酸酯钠对HK-2细胞的影响无明显差异;HK-2细胞染毒24h后,细胞凋亡和胀亡均明显增多,2、4μl/mL草鱼胆汁和66.7、133.4μg/mL鲤醇硫酸酯钠导致细胞凋亡样坏死明显增多,而8μl/mL草鱼胆汁和533.6μg/mL鲤醇硫酸酯钠导致细胞胀亡样坏死明显增多。结论中、小剂量的草鱼胆汁毒素主要导致HK-2细胞呈现凋亡样坏死,大剂量的草鱼胆汁毒素主要导致HK-2细胞呈现胀亡样坏死;鲤醇硫酸酯钠是草鱼胆汁毒素的主要毒性成分。

Objective To observe the death mode of human proximal tubular epithelial cell line（HK-2） exposed to gass carp bile and its extract（sodium cyprinol sulphate,SCS）.Methods SCS was extracted from the grass carp bile according to method of the literature reported.HK-2 was cultured in vitro.MTT assay was used to evaluate the multiplication-inhibiting of grass carp bile and SCS on HK-2 cells.Flow cytometry was used to determine the main death way of HK-2 cells exposed to different concentrations of grass carp bile and SCS.Results The viability of HK-2 cells induced by grass carp bile and SCS was significantly reduced.The viability of cells was concentration and time dependant,and not different between rass carp bile and SCS groups.After treatment with a series concentrations of grass carp bile and SDS for 24 hours,apoptosis and oncosis of the cultured HK-2 cells which based on the experimental datas of flow cytometry were obviously increased,and increased apoptosis cells was mainly found in 2,4μl/ml grass carp bile and 66.7,133.4μg/ml SCS,but increased oncosis cells in 8μl/ml grass carp bile and 533.6μg/ml SCS.Conclusions The middle and small dose grass carp bile and SCS result mainly in apoptosis of the HK-2 cells,while the large dose is mainly oncosis.SDS is one of the essential virulent components of grass carp bile.