他克莫司对清蛋白超载的大鼠肾小管上皮细胞凋亡的影响

Inhibitory effect of FK506 on d-BSA-induced apoptosis of NRK-52E cells

目的蛋白尿不仅是难治性肾病重要的临床表现,其主要成分清蛋白是引起肾小管上皮细胞凋亡的重要原因。文中探讨他克莫司(tacrolimus,FK506)对去脂牛血清清蛋白(denatured bovine serum albumin,d-BSA)超载的大鼠肾小管上皮细胞(normal rat kidney cell line,NRK-52E)凋亡率的影响。方法将不同浓度FK506于不同时间作用于d-BSA超载的NRK-52E细胞,用流式细胞术检测NRK-52E细胞凋亡率的情况。结果①与对照组相比,终质量浓度为1 mg/ml的d-BSA对NRK-52E细胞无明显促进凋亡作用,且与d-BSA孵育时间无明显关系(P>0.05);终质量浓度为5、10、20、50 mg/ml d-BSA干预NRK-52E细胞24 h可上调NRK-52E细胞的凋亡率,凋亡率随着d-BSA浓度的增加呈上升期趋势(P<0.05)。②终质量浓度为0.1 ng/ml的FK506预处理NRK-52E细胞4 h后与不同浓度d-BSA共孵育引起的细胞凋亡率与对照组无异,且与共孵育时间无明显关系(P>0.05)。终质量浓度为1、10、20 ng/ml的FK506预处理4 h后与20 mg/ml d-BSA共孵育6 h,具有降低细胞凋亡率的作用,并呈现与FK506浓度与孵育时间的依赖关系(P<0.01)。结论 d-BSA超载NRK-52E细胞可以促进细胞凋亡;FK506具有抑制d-BSA促凋亡的作用。

Objective The purpose of this study was to investigate effect of FKS06 on d-BSA-induced apoptosis of normal rat kidney （NRK）-52E cells. Methods We exposed d-BSA-induced NRK-52E cells to different concentrations of FKS06 for different periods of time, and then determined the apoptosis of the cells by flow cytometry. Results d-BSA at 1 mg/ml produced no obvious difference on the apoptosis of the NRK-52E cells, and it was not significantly related with time （P 〉 0.05）. Exposure to d-BSA at 5, 10, 20 and 50 mg/ml for 24 hours markedly increased the apoptosis of the NRK-52E cells in a dose-dependent manner （P 〈 0.05 ）. Pretreatment of NRK-52E with FKS06 at 0.1 ng/ml for 4 hours produced no significant difference in d-BSA-induced apoptosis of the NRK-52E cells, and it showed no significant correlation with time （P 〉 0.05）. Six hours of co-incubation with d-BSA after pretreat- ment with FKS06 at 1, 10 and 20 ng/ml for 4 hours remarkably inhibited the apoptosis of the the NRK-52E cells in a dose- and time- dependent manner （P 〈 0.01 ）. Conclusion D-BSA may induce the apoptosis of NRK-52E ceils, and FKS06 can inhibit it.