整合素连接激酶在衰老鼠肾间质损害中的表达及意义

Expression and role of integrin-linking kinase in renal tubulointerstitial lesion of aging rats

目的探讨整合素连接激酶(ILK)在衰老鼠肾间质损害中的表达和意义。方法将3月龄、26月龄Wistar大鼠制成单侧输尿管梗阻(UUO)模型,用免疫荧光、Westernblot、RT-PCR等方法动态观察不同鼠龄间ILK、纤维连接蛋白(Fn)的mRNA和蛋白在UUO术前及术后3、7、14d表达的变化。结果随UUO术后时间的延长,各月龄鼠肾间质相对面积及肾小管间质纤维化面积明显增加(P<0·01),Fn、ILK的mRNA和蛋白表达也增加(P<0·01);26月龄与3月龄UUO大鼠ILKmRNA半定量值在术后3d为0·98±0·06、0·72±0·06,术后14d为1·49±0·05、1·03±0·04;ILK蛋白半定量值在术后3d为0·57±0·04、0·52±0·03,术后14d为0·76±0·04、0·63±0·03。随UUO术后时间的延长,ILK的mRNA和蛋白表达水平与肾间质相对面积的改变呈正相关(r分别为0·71、0·80,P值均小于0·05)。结论ILK的表达随鼠龄的增加及UUO术后时间的延长呈进行性增加,其高表达可能是促进肾脏纤维化和肾脏衰老的原因之一。

Objective To explore the expression and role of integrin-linking kinase（ILK） in aging rats subjected to unilateral ureteral obstruction （UUO）. Methods The UUO model and sham-operated rats （SHAM） were established with 3-month-old rats and 26-month-old rats and the rats were sacrificed before UUO and after UUO at 3, 7 and 14 days. Immunofluorescent staining, Western blot, RT-PCR, etc were applied to detect the expressions of ILK and fibronectin（FN） in the renal of UUO rat at each time point. Rusults With the time of UUO, the relative interstitial areas and tubulointerstitial fibrotic areas in 26- month-old rats were significantly higher than those in 3-month-old ones at each time point（P 〈 0. 01 ）and the mRNA and protein levels of ILK and FN are increasing with the time of UUO in both groups （ P 〈 0.01 ）. ILK mRNA semi-quantitative values of 26-month and 3-month rats were 0. 98 ＋ 0. 06 vs 0. 72 ±0. 06, 3 days after UUO （P〈0.01）, 1.49 ±0.05 vs 1.03 ±0.04, 14 days after UUO （P 〈0.01）, respectively. ILK proteinsemi-quantitative values of 26-month and 3-month rats were 0. 57 ± 0. 04 vs 0. 52 ±0. 03, 3 days after UUO （P〈0.01）, 0.76 ±0.04 vs 0.63 ±0.03, 14 days after UUO （P 〈0.01）, respectively. The expressions of ILK mRNA and protein level were positively correlated with the relatively renal interstitial areas （r = 0. 71, P 〈 0. 05 and r = 0. 80, P 〈 0. 05, respectively）. And the whole levels of ILK and FN in 26-month-old rats are higher than those of 3-month-old rats. Conclusion With aging, the expressions of ILK and FN increase progressively in UUO rat, and the over-expression of ILK probably promotes renal tubulointerstitial fibrosis and aging.