Dual Effects of Bilirubin on the Proliferation of Rat Renal NRK52E Cells and its Association with Gap Junctions
Dual Effects of Bilirubin on the Proliferation of Rat Renal NRK52E Cells and its Association with Gap Junctions
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DOI：
10.2203/dose-response.12-003.hei
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发表时间：
2013-04
期刊：
影响因子：
2.5
通讯作者：
Yanling Wang;Qiong-fang Zhu;Chen-fang Luo;Ailan Zhang;Z. Hei;G. Su;Z. Xia;M. Irwin
中科院分区：
文献类型：
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作者：
Yanling Wang;Qiong-fang Zhu;Chen-fang Luo;Ailan Zhang;Z. Hei;G. Su;Z. Xia;M. Irwin
目的胆红素对肾脏病理生理学的影响存在争议。本研究旨在观察胆红素对正常大鼠肾小管上皮细胞株（NRK 52 E）增殖的影响及其与缝隙连接功能的关系。方法将NRK 52 E细胞高、低密度接种，加入不同浓度的胆红素作用24 h。通过测量细胞活力和增殖来评估细胞损伤，并且通过降落伞染料偶联测定来评估间隙连接功能。通过Western印迹法评估连接蛋白43蛋白。结果17.1 ~ 513μmol/L浓度范围内，胆红素剂量依赖性地增加高密度和低密度接种细胞的存活率和集落形成率（与溶剂组相比，p均<0.05），并增加高密度细胞的细胞间荧光透射和Cx43蛋白表达。当BR浓度从684 μmol/L增加到1026μmol/L时，上述作用逐渐被逆转。在高密度细胞中，间隙连接抑制剂12-O-十四酰佛波醇13-乙酸酯减弱了藜芦醇诱导的集落形成和荧光透射的增强。然而，在高浓度胆红素（1026μmol/L）存在下，维甲酸激活缝隙连接降低集落形成率。结论胆红素对NRK 52 E细胞增殖具有双向作用，其作用机制可能与影响细胞间隙连接功能有关。
Objective The effect of bilirubin on renal pathophysiology is controversial. This study aimed to observe the effects of bilirubin on the proliferation of normal rat renal tubular epithelial cell line (NRK52E) and its potential interplay with gap junction function. Methods Cultured NRK52E cells, seeded respectively at high- or low- densities, were treated with varying concentrations of bilirubin for 24 hours. Cell injury was assessed by measuring cell viability and proliferation, and gap junction function was assessed by Parachute dye-coupling assay. Connexin 43 protein was assessed by Western blotting. Results At doses from 17.1 to 513μmol/L, bilirubin dose-dependently enhanced cell viability and colony-formation rates when cells were seeded at either high- or low- densities (all p<0.05 vs. solvent group) accompanied with enhanced intercellular fluorescence transmission and increased Cx43 protein expression in high-density cells. However, the above effects of BR were gradually reversed when its concentration increased from 684 to 1026μmol/L. In high-density cells, gap junction inhibitor 12-O-tetradecanoylphorbol 13-acetate attenuated bilirubin-induced enhancement of colony-formation and fluorescence transmission. However, in the presence of high concentration bilirubin (1026μmol/L), activation of gap junction with retinoid acid decreased colony-formation rates. Conclusion Bilirubin can confer biphasic effects on renal NRK52E cell proliferation potentially by differentially affecting gap junction functions.