反向模式钠钙离子交换体抑制剂降低造影剂诱导的肾小管上皮细胞凋亡

目的 探讨反向模式钠钙离子交换体抑制剂KB-R7943是否对造影剂肾损伤具有保护作用.方法 培养大鼠肾小管上皮细胞分别与不同浓度KB-R7943(10-5,10-6 mol/L)作用12 h后,加入造影剂作用1 h.采用LDH检测细胞损伤,倒置显微镜观察细胞形态变化,流式细胞仪检测细胞凋亡,共聚焦显微镜测定细胞内钙和反应氧产物水平,RT-PCR检测钠钙离子交换体mRNA表达.相同渗透压甘露醇作对照.数据以均数±标准差(x±s)表示,统计采用方差分析和q检验,简单直线相关分析两者相关性,以P<0.05为差异具有统计学意义.结果 造影剂作用1 h诱导了明显细胞损伤和细胞凋亡,细胞内钙和反应氧产物增加,KB-R7943降低了细胞内钙和反应氧产物水平,同时降低了细胞损伤和细胞凋亡并呈剂量效应;钠钙离子交换体mRNA表达无变化.结论 KB-R794对造影剂诱导的肾小管上皮细胞损伤具有保护作用.

Reverse mode of Na+/Ca2+ exchange inhibitor, KB-r7943 attenuates tubular epithelial cell apoptosis inducde by contrast media

Objective To investigate the renoprotective effects of KB-R7943 on renal injury induced bycontrast media. Method Tubular epithelial cells were trested with varions cencentration of KB-B7943 (10-5,10-6 mol/L) for 12 hours before contrast media was used. After cells we, re incubated with contrast media (CM)(110 mgI/L) for 1 hour, cells injury was assessed by using LDH, and cell morphologic changes and cells apopto-sis were evaluated with inverted microscope and flow cytometry, respoelively. Intracellular Ca2+ and reactive oxy-gen species (ROS) were analyzed by using confocal microscope. The expression of Na+/Ca2+exchanger mRNAwas evaluated by RT-PCR. Mannitol with same osmolarity (20% mannitol, 770 mOsm/L) of CM was used as con-trol. One-way analysis of variance and q-test were used for comparison between groups. The simple linear correla-tion was employed to analyze the correlation. P < 0.05 was considered significant. Results Contrast media sig-nificantly induced tubular cells damage significantly and apoptosis at 1 hour after incubation, meanwhile, intracel-lular Ca2+ and ROS were inoreased progressively in CM group. KB-R7943 significantly attenuated cells damage andapoptosis in dose-dependent in association with decreased intracellular Ca2+ and ROS. Expression of Na+/Ca2+exchanger mRNA was not changed. Conclusions KB-R7943 has renoprotective effects on the contrast-media-in-duced renal tubular cyotoxicity