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目的探讨白蛋白诱导肾小管上皮细胞凋亡以及诱导凋亡的信号传导机制。方法将培养的大鼠肾小管细胞NRK-52E分别与不同浓度(10、20、30mg/ml)的去脂无内毒素牛血清白蛋白(BSA)共同孵育6、12、18和24h。透射电镜、共聚焦激光显微镜和流式细胞仪检测细胞凋亡。BSA20mg/ml刺激NRK-52E细胞15、30、60和120min后,Westen印迹测定p38、氨基末端激酶(JNK)和细胞外信号调节激酶(ERK)活性。将SB202190(20μmol/L,p38抑制剂)、SP600125(10μmol/L,JNK抑制剂)和PD98059(20μmol/L,ERK抑制剂)分别与白蛋白和NRK-52E细胞共同孵育24h后检测细胞凋亡。结果白蛋白以时间和剂量依赖方式诱导肾小管细胞凋亡。白蛋白与NRK-52E细胞共孵育后,p38和JNK活性明显升高,ERK活性显著降低。SB202190和SP600125可分别抑制白蛋白诱导NRK-52E细胞凋亡,而PD98059促进白蛋白诱导的NRK-52E细胞凋亡。结论白蛋白以时间和剂量依赖方式诱导肾小管细胞凋亡,而p38和JNK激活与ERK抑制介导了白蛋白诱导的肾小管细胞凋亡。
Objective To investigate the signaling pathway induced by albumin in renal tubular epithelial cells and to induce apoptosis. Methods Cultured rat renal tubular cells NRK-52E were incubated with different concentrations of defatted endotoxin-free bovine serum albumin (BSA) at different concentrations (10, 20 and 30 mg / ml) for 6, 12, 18 and 24 h. Transmission electron microscopy, confocal laser scanning microscopy and flow cytometry were used to detect apoptosis. After 15, 30, 60 and 120 min stimulation of NRK-52E cells with BSA 20 mg / ml, p38, N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) activity were determined by Westen blotting. Cell apoptosis was detected by incubating SB202190 (20μmol / L, p38 inhibitor), SP600125 (10μmol / L, JNK inhibitor) and PD98059 (20μmol / L, ERK inhibitor) with albumin and NRK- . Results Albumin induced tubular cell apoptosis in a time and dose dependent manner. After incubation of albumin and NRK-52E cells, the activity of p38 and JNK was significantly increased and ERK activity was significantly decreased. SB202190 and SP600125 respectively inhibited albumin-induced apoptosis in NRK-52E cells, whereas PD98059 promoted albumin-induced apoptosis in NRK-52E cells. Conclusion Albumin induced apoptosis of tubular cells in a time-and dose-dependent manner, while p38 and JNK activation and ERK inhibition mediated albumin-induced tubular cell apoptosis.