Liu Qing-juan, Shi Yong-hong, Liu Shu-xia, Hao Jun, Cao Yan-ping, Li Hang, Wei Jin-ying, Duan Hui-jun
Department of Pathology, Hebei Medical University, Shijiazhuang 050017, Hebei, China.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2010 Dec;22(12):747-9.
To investigate the effect of AG490, a Janus kinase 2 inhibitor, on epithelial-myofibroblast transdifferentiation induced by interleukin-1β (IL-1β).
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).
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 statistically 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.
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.
研究Janus激酶2抑制剂AG490对白细胞介素-1β（IL-1β）诱导的上皮-肌成纤维细胞转分化的影响。
将培养的人肾小管上皮细胞系（HKCs）分为三组：空白对照组、IL-1β（5 ng/ml）组和AG490组（IL-1β 5 ng/ml + AG490 10 μmol/L）。干预后24、48、72小时收集各组细胞。采用免疫细胞化学和蛋白质印迹分析检测细胞角蛋白-18（CK-18）和α-平滑肌肌动蛋白（α-SMA）的表达。
空白对照组CK-18表达较高（1.25±0.08），α-SMA表达较弱（0.17±0.01）。在IL-1β组，随着刺激时间延长，CK-18蛋白水平逐渐降低（24小时：0.69±0.04，48小时：0.52±0.03，72小时：0.30±0.01），而α-SMA表达水平逐渐升高（24小时：0.56±0.04，48小时：1.05±0.07，72小时：1.43±0.07），空白对照组与IL-1β组之间差异有统计学意义（均P<0.05）。给予AG490可恢复CK-18的表达（24小时：1.07±0.07，48小时：0.93±0.06，72小时：0.83±0.06），并抑制IL-1β诱导的α-SMA表达（24小时：0.33±0.01，48小时：0.52±0.01，72小时：0.61±0.04）。AG490组与IL-1β组之间差异有统计学意义（均P<0.05）。免疫细胞化学结果与蛋白质印迹结果一致。
IL-1β可诱导肾小管上皮细胞转分化，上调α-SMA表达，促使肾小管上皮细胞向肌成纤维细胞转化，而AG490可抑制IL-1β的作用。
