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目的:探讨他克莫司(tacrolimus,FK506)对去脂牛血清白蛋白(denatured bovine serum albumin,d-BSA)超载的大鼠肾小管上皮细胞(normal rat kidney proximal tubular epithelial cell line,NRK-52E)增殖的影响。方法:(1)以不同终质量浓度(1、5、10、20、50 mg·ml-1)d-BSA超载NRK-52E,以正常培养的NRK-52E细胞为对照组,观察不同浓度d-BSA对细胞增殖的影响及d-BSA(20 mg·ml-1)超载不同时间(6、12、24 h)对NRK-52E增殖的影响。(2)以不同浓度(0.1、1、10、20 ng·ml-1)FK506预处理NRK-52E 4 h后加入终质量浓度为20 mg·ml-1d-BSA共孵育24 h,观察不同浓度FK506预处理对d-BSA超载细胞增殖的影响。(3)以10 ng·ml-1的FK506预处理4 h后加入终质量浓度为20 mg·ml-1d-BSA共孵育NRK-52E,观察不同时间(6、12、24 h)对细胞增殖影响。以上细胞增殖测定均采用四甲基偶氮唑盐法(MTT)。结果:(1)终质量浓度为1 mg·ml-1的d-BSA超载NRK-52E 24 h后无明显促进细胞增殖的作用(P>0.05);终质量浓度为5 mg·ml-1的d-BSA能够促进细胞增殖(P<0.05),终质量浓度为10、20、50 mg·ml-1的d-BSA能够显著促进细胞增殖(P<0.01)。(2)以终质量浓度为0.1 ng·ml-1的FK506预处理NRK-52E,细胞增殖与对照组比较差异无统计学意义(P>0.05);终质量浓度为1 ng·ml-1的FK506具有抑制d-BSA诱导的细胞增殖的作用(P<0.05);10、20 ng·ml-1FK506能够显著抑制-d-BSA诱导的细胞增殖(P<0.01)。(3)终质量浓度为10 ng·ml-1的FK506预处理4 h后加入终质量浓度为20 mg·ml-1d-BSA共孵育NRK-52E,6 h后,具有抑制细胞增殖的作用(P<0.05),共孵育12、24 h后,抑制细胞增殖的作用更加显著(P<0.01)。结论:d-BSA超载NRK-52E可以诱导细胞增殖,FK506具有抑制d-BSA促进NRK-52E细胞增殖的作用。
OBJECTIVE: To investigate the effect of tacrolimus (FK506) on the proliferation of rat renal tubular epithelial cell line (NRK-52E) loaded with denatured bovine serum albumin (d-BSA) ) Proliferation. Methods: (1) NRK-52E cells were overexpressed by d-BSA at different final concentrations (1,5,10,20,50 mg · ml-1), and NRK-52E cells cultured in normal conditions were used as control. -BSA on cell proliferation and the effect of d-BSA (20 mg · ml-1) on the proliferation of NRK-52E at different times (6,12,24 h). (2) NRK-52E cells were pretreated with FK506 at different concentrations (0.1, 1, 10 and 20 ng · ml-1) for 4 h and then incubated with 20 mg · ml-1d-BSA for 24 h. Effect of FK506 preconditioning on the proliferation of d-BSA overload cells. (3) After pretreatment with FK506 at 10 ng · ml-1 for 4 h, NRK-52E was co-incubated with a final concentration of 20 mg · ml-1d-BSA, and the cell proliferation was observed at different times (6,12,24 h) influences. The above cell proliferation assay using tetramethylazolamide method (MTT). RESULTS: (1) NRK-52E over-loaded with d-BSA at a final concentration of 1 mg · ml-1 did not significantly promote cell proliferation (P> 0.05) at a final concentration of 5 mg · ml-1 d-BSA can promote cell proliferation (P <0.05), and d-BSA at the final concentration of 10,20,50 mg · ml-1 could significantly promote cell proliferation (P <0.01). (2) NRK-52E was pretreated with FK506 at a final concentration of 0.1 ng · ml-1, and there was no significant difference in cell proliferation between the two groups (P> 0.05); the final concentration was 1 ng · ml-1 FK506 could inhibit the proliferation of cells induced by d-BSA (P <0.05); 10,20 ng · ml-1 FK506 could significantly inhibit the proliferation of cells induced by d-BSA (P <0.01). (3) After pretreatment with FK506 at a final concentration of 10 ng · ml-1 for 4 h, NRK-52E was incubated at a final concentration of 20 mg · ml-1d-BSA for 6 h and then inhibited cell proliferation ( P <0.05). After incubated for 12 and 24 h, the effect of inhibiting cell proliferation was more significant (P <0.01). CONCLUSION: Over-loaded NRK-52E can induce cell proliferation, and FK506 can inhibit the proliferation of NRK-52E cells induced by d-BSA.