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Dual effects of bilirubin on the proliferation of rat renal NRK52E cells and its association with gap junctions

机译：胆红素对大鼠肾脏NRK52E细胞增殖及其与间隙连接的关联的双重作用

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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.
机译：目的：胆红素对肾脏病理生理的影响尚存争议。本研究旨在观察胆红素对正常大鼠肾小管上皮细胞系（NRK52E）增殖及其与间隙连接功能的潜在相互作用的影响。方法：分别以高密度或低密度接种的培养的NRK52E细胞用不同浓度的胆红素处理24小时。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。通过蛋白质印迹评估连接蛋白43蛋白。结果：在17.1至513μmol/ L的剂量下，以高密度或低密度（相对于溶剂组而言，所有p <0.05）接种细胞时，胆红素剂量依赖性地提高了细胞活力和集落形成率，同时增强了细胞间荧光在高密度细胞中传播和Cx43蛋白表达增加。然而，当BR的浓度从684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-十四烷酰佛波醇13-乙酸盐减弱了胆红素诱导的集落形成和荧光传递的增强。然而，在高浓度胆红素（1026μmol/ L）的存在下，类视黄酸间隙连接的激活降低了菌落形成率。结论：胆红素可能通过差异影响间隙连接功能而对肾脏NRK52E细胞增殖具有双相作用。

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