Yao Luyu, Kolluru Gopi K, Kevil Christopher G, Zhang Wayne W
1Department of Surgery, Louisiana State University Health Sciences Center, Shreveport, LA, USA.
Vasc Endovascular Surg. 2013 Nov;47(8):632-8. doi: 10.1177/1538574413503560. Epub 2013 Sep 4.
To investigate the effect of Iodixanol on kidney proximal tubular cell line human kidney 2 (HK-2).
The HK-2 cells were treated with Iodixanol. A Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay was used to evaluate apoptosis. Cell viability was measured by proliferation assay kit. Cell permeability changes were assessed by transwell assay and intercellular gaps measurement. Expression of claudin-2 was assessed by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and Western blot.
Iodixanol reduced tubule cell viability (P < .01) but did not cause apoptosis. The intercellular gap formation (P < .01) and transwell (P < .05) assays revealed that cell permeability significantly increased after Iodixanol treatment of monolayer cells. Western blot and qRT-PCR showed significant upregulation of claudin-2 protein (P < .05) and messenger RNA expression (P < .01).
Our in vitro data do not support the hypothesis that direct kidney cell death from Iodixanol is a major mechanism of contrast-induced nephropathy (CIN). However, increased permeability of proximal tubule epithelium caused by Iodixanol may play an important role in CIN.
研究碘克沙醇对人近端肾小管上皮细胞系（HK-2）的影响。
用碘克沙醇处理HK-2细胞。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL法）评估细胞凋亡。使用增殖检测试剂盒测量细胞活力。通过Transwell实验和细胞间隙测量评估细胞通透性变化。采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测claudin-2的表达。
碘克沙醇降低了肾小管细胞活力（P < 0.01），但未引起细胞凋亡。细胞间隙形成实验（P < 0.01）和Transwell实验（P < 0.05）表明，碘克沙醇处理单层细胞后细胞通透性显著增加。蛋白质免疫印迹法和qRT-PCR显示claudin-2蛋白表达显著上调（P < 0.05），信使核糖核酸表达显著上调（P < 0.01）。
我们的体外实验数据不支持碘克沙醇直接导致肾细胞死亡是造影剂肾病（CIN）主要机制的假说。然而，碘克沙醇引起的近端肾小管上皮细胞通透性增加可能在CIN中起重要作用。
