钠钙离子交换体介导的钙超载对造影剂诱导NRK52E细胞损伤的影响  

Effects of Ca^(2+) Overload via Na^+/Ca^(2+) Exchanger on Renal Tubular Cell Apoptosis Induced by Contrast Media

作　　者：杨定平[1] 贾汝汉[1] 丁国华[1] 杨红霞[1] 

机构地区：[1]武汉大学人民医院肾内科,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2008年第5期579-583,共5页Medical Journal of Wuhan University

基　　金：湖北省自然科学基金资助项目(编号:2007ABA254)

摘　　要：目的:本研究探讨钠钙离子交换体(NCX)是否参与了造影剂诱导的肾小管上皮细胞损伤及反向模式钠钙离子交换体抑制剂KB-R7943对造影剂诱导的肾小管上皮损伤的影响。方法:鼠肾小管上皮细胞(NRK52E)被分成6组:正常对照组、造影剂组、甘露醇组、钙离子拮抗剂(CCB)组、KB-R7943(10-5mol/L)和KB-R7943(10-6mol/L)组。造影剂作用1 h后,细胞损伤采用LDH检测,细胞形态变化及细胞凋亡分别由倒置显微镜和流式细胞仪检测;细胞内钙采用共聚焦微镜测定;钠钙离子交换体mRNA表达采用RT-PCR测定。结果:造影剂作用1 h后,细胞损伤、细胞凋亡、细胞内钙进行性增加,显著高于相同渗透压甘露醇组;KB-R7943显著改善了上述异常,且较相同浓度CCB具有更好的效果并显示出剂量效应;钠钙离子交换体mRNA表达没有变化。结论:经反向模式钠钙离子交换体导致的细胞内钙超载参与了造影剂诱导的肾小管上皮细胞损伤;反向模式钠钙离子交换体抑制剂KB-R7943以呈剂量方式对造影剂诱导的肾小管上皮细胞损伤发挥保护作用。Objective： To investigate whether intracellular Ca^2＋ overload via Na＋/Ca^2＋ exchanger （NCX） system contributes to the contrast media nephrotoxicity and whether KB-R7943, an inhibitor of reverse mode of Na＋/Ca^2＋ exchanger, and attenuates contrast media-induced tubular cell injury. Methods： The tubular cells were treated with KB-R7943 at various concentrations at 12 h before contrast media administration. Lactic dehydrogenase （LDH） was assayed to evaluate the cell damage level, which was measured by automatic biochemistry analyzer. Flow cytometry was used to quantify the apoptotic cells. Morphologic changes of tubular cells was observed by inverted microscope, and intracellular Ca^2＋ concentration was detected by fluorescence probe using confocal microscope. The expression of Na^＋/Ca^2＋ exchanger mRNA was evaluated by reverse transcription-polymerase chain reaction. Results： Contrast media induced I.DH level to in- crease significantly in the culture media and apoptotic tubular ceils. Intracellular Ca^2＋ concentra- tion also increased progressively following contrast media administration. KB-R7943 attenuated the cell injury and decreased the tubular cell apoptosis induced by contrast media through decrea sing intracellular Ca^2＋ overload. The expression of Na^＋/Ca^2＋exchanger mRNA had no change at 30 min and 60 min after contrast media administration. Conclusion. Intracellular Ca2＋ overload via the NCX system contributes to the contrast media induced nephrotoxieity. KB-R7943 exerted renoprotective effects through inhibiting intracellular Ca^2＋ overload.

关 键 词：造影剂 肾小管上皮细胞 钠钙离子交换体 钙超载 

分 类 号：R981.1[医药卫生—药品]