摘要目的 探讨Janus激酶2抑制剂AG490对白细胞介素-1β(IL-1β)诱导人肾小管上皮细胞转分化的影响. 方法 将体外培养人肾近曲小管上皮细胞株(HKCs)分为空白对照组、IL-1β(5 ng/ml)刺激组及AG490干预组(IL-1β 5 ng/ml+AG490 10 μmol/L).分别于处理后24、48、72 h收集细胞,采用免疫细胞化学染色和蛋白质免疫印迹法(Western blotting)检测细胞角蛋白-18(CK-18)、α-平滑肌肌动蛋白(α-SMA)蛋白表达. 结果 正常肾小管上皮细胞内其自身标志物CK-18高表达(1.25±0.08),而α-SMA表达很少(0.17±0.01).IL-1β刺激组随刺激时间延长CK-18表达逐渐减少(24 h:0.69±0.04,48 h:0.52±0.03,72 h:0.30±0.01),而α-SMA蛋白合成逐渐增加(24 h:0.56±0.04,48 h:1.05±0.07,72 h:1.43±0.07),与空白对照组比较差异均有统计学意义(均P＜0.05).AG490干预能使CK-18的表达逐渐恢复(24 h:1.07±0.07,48 h:0.93±0.06,72 h:0.83±0.06),并减弱IL-1β对肾小管上皮细胞α-SMA蛋白的诱导作用(24 h:0.33±0.01,48 h:0.52±0.01,72 h:0.61±0.04),且作用显著(均P＜0.05).免疫细胞化学染色观察结果与其一致. 结论 炎症因子IL-1β通过降低肾小管上皮细胞CK-18表达,上调α-SMA表达,促使肾小管上皮细胞转分化为肌成纤维细胞;Janus激酶2抑制剂AG490能部分阻断IL-1β对肾小管上皮细胞的促转分化作用.

abstractsObjective To investigate the effect of AG490,a Janus kinase 2 inhibitor,on epithelial-myofibroblast transdifferentiation induced by interleukin-1β(IL-1β). Methods Cultured human renal tubular epithelial cell line(HKCs) were divided into three groups:blank control group,IL-1β(5 ng/ml) group and AG490 group(IL-1β 5 ng/ml+AG490 10 μmol/L).The cells in all groups were collected at 24,48,72 hours after intervention.Immunocytochemistry and Western blotting analysis were used to determine the expressions of cytokeratin-18(CK-18) and α-smooth muscle actin(α-SMA). Results The higher expression of CK-18(1.25±0.08) and mild expression of α-SMA(0.17±0.01) were found in blank control group.In IL-1β group,the protein level of CK-18 was gradually decreased with prolongation of stimulus(24 hours:0.69±0.04,48 hours:0.52±0.03,72 hours:0.30±0.01),while the expression level of α-SMA was gradually increased(24 hours:0.56±0.04,48 hours:1.05±0.07,72 hours:1.43±0.07),and the difference between blank control group and IL-1β group was statiscally significant(all P<0.05).The administration of AG490 could restore the expression of CK-18(24 hours:1.07±0.07,48 hours:0.93±0.06,72 hours:0.83±0.06),and inhibit the expression of α-SMA induced by IL-1β(24 hours:0.33±0.01,48 hours:0.52±0.01,72 hours:0.61±0.04).There was significant difference between AG490 group and IL-1β group(all P<0.05).The results of immunocytochemistry and that of Western blotting were identical. Conclusion IL-1β can induce the transdifferentiation of renal tubular epithelial cells,up-regulate the expression of α-SMA,induce the renal tubular epithelial cells to transform to myofibroblast,while AG490 can inhibit the effect of IL-1β.