KB-R7943对造影剂诱导NRK52E细胞凋亡及p38MAPK信号通路的影响  

KB-R7943 decreased renal tubular cell apoptosis and p38 expression Induced by contrast media

作　　者：杨定平[1] 贾汝汉[1] 丁国华[1] 张杰[1] 

机构地区：[1]武汉大学人民医院泌尿疾病研究所,430060

出　　处：《中华实验外科杂志》2008年第5期641-643,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省自然科学基金资助项目（2007ABA254）

摘　　要：目的探讨经钠钙离子交换体介导的细胞内钙超载及p38MAPK信号通路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细胞凋亡及p38MAPK信号通路的影响。方法鼠肾小管上皮细胞（NRK52E）被分成6组：A正常对照组；B造影剂组；C甘露醇组；DCCB（1×10^-5mol／L）；EKB—R7943（1×10^-5mol／L）；FKB—R7943（1×10^-6mol／L）。造影剂作用1h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；细胞内钙通过Fluo-4染色共聚焦微镜测定；钠钙离子交换体mRNA表达采用RT—PCR测定；p38蛋白的表达采用Westernblot测定。结果造影剂作用1h后，细胞损伤、细胞凋亡、细胞内钙明显增加，显著高于相同渗透压甘露醇组；p-p38表达30min时增加、60min时下降。KB—R7943显著降低细胞损伤、细胞凋亡、细胞内钙水平、抑制p—p38的高表达述并显示出剂量效应；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诱导了造影剂诱导的肾小管上皮细胞凋亡及p-p38蛋白的高表达；KB—R7943以呈剂量方式降低造影剂诱导的肾小管上皮细胞凋亡及p—p38蛋白的表达。Objective To investigate whether intracellular Ca^2＋ overload via NCX system and p38MAPK contributes to the contrast media nephrotoxicity and whether KB-R7943 ,an inhibitor of reverse mode of Na^＋/Ca^2＋ exchanger, attenuates contrast media-induced tubular cell injury and inhibits the expression of p-p38. Methods The tubular cells were treated with KB-R7943 at various concentrations 1 h before contrast media administration. Lactic dehydrogenase （LDH） was determined to evaluate the cell damage severity. Flow cytometry was used to quantify the apoptotic cells. Morphologic change of tubular cell was observed by inverted microscope and intracellular Ca^2＋ concentration was detected by fluorescence probe using confocal microscope. The expression of p-p38 protein was examined by Western blot. The expression of Na^＋/Ca^2＋ exchanger mRNA was evaluated by reverse transcription-polymerase chain reaction. Results Contrast media induced significantly increased LDH level in the culture media and apoptotic tubular cells. Intracellular Ca^2＋ concentration was also increased progressively following contrast media administration. The expression of p-p38 protein was increased significantly at 30 min and decreased at 60 min in contrast media group. The expression of Na^＋/Ca^2＋ exchanger mRNA had no change at 30 min and 60 min. KB-R7943 attenuated the cell injury, decreased the tubular cell apoptosis and inhibited the increased expression of p-p38 protein induced by contrast media through decreasing intracellular Ca^2＋ overload. Conclusion Intracellular Ca^2＋ overload via the NCX system contributed to the contrast media-in- duced nephrotoxicity;KB-R7943 exerted renoprotective effects through decreasing intracellular Ca^2＋ over- load and inhibiting the expression of p-p38 protein.

关 键 词：造影剂 肾小管上皮细胞 P38MAPK 

分 类 号：R692.9[医药卫生—泌尿科学]