AG490对肾小管上皮细胞转分化影响的实验研究

目的探讨炎症介质的特异性阻断剂AG490在白介素-1β(interleukin-1βIL-1β)诱导的高糖培养的人肾小管上皮细胞转分化中的作用。方法体外培养人肾近曲小管上皮细胞株(HKCs),随机分为高糖对照组(30mmol·L-1);高糖(30mmol·L-1)+IL-1β(5μg·L-1)组;高糖(30mmol·L-1)+IL-1β(5μg·L-1)+AG490(10μmol·L-1)组。分别于处理后24、48、72h收集细胞,采用免疫细胞化学染色和蛋白Western blot法检测细胞角蛋白-18(cytokeratin-18,CK-18)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)水平。结果IL-1β能使高糖培养的人肾小管上皮细胞α-SMA蛋白的合成增加,而肾小管上皮细胞的标志物CK-18的表达逐渐减少;IL-1β刺激的同时加入AG490干预能减弱IL-1β对肾小管上皮细胞α-SMA蛋白的诱导作用,并使CK-18的表达回升。结论炎症因子IL-1β能促进高糖培养的HKC发生表型转化,而炎症特异性阻断剂AG490能部分阻断IL-1β的该作用。

Experimental study about the role of AG490 on tubular epithelial- myofibroblast transdifferentiation

Aim To investigate the role of AG490 on tubular epithelial-myofibroblast transdifferentiation induced by interleukin-1β and high glucose.Methods Cultured HKCs were divided into three groups:high glucose(30 mmol·L-1) group,high glucose(30 mmol·L-1)+IL-1β(5 μg·L-1) group,and high glu-cose(30 mmol·L-1)+IL-1β(5 μg·L-1)+AG490(10 μmol·L-1) group.Cells were collected after cultured 24,48,72 hours.Immunocytochemistry and Western blot analysis were used to determine the expression of cytokeratin-18(CK-18) and α-smooth muscle actin (α-SMA).Results The protein level of α-SMA in tubular epithelial cells induced by high glucose and IL-1β was higher than that stimulated by high glucose alone,while the expression of CK-18 was markedly decreased.Administration of AG490 could decrease the expression of α-SMA and increase the expression of CK-18.Conclusion AG490 can inhibit the transdifferentiation of renal tubular epithelial cells induced by high glucose and IL-1β.