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目的:观察终末期糖基化终产物(AGEs)对正常大鼠近端肾小管上皮细胞(NRK52E)转分化、colla-genⅠ合成及Sm ads信号通路的影响。方法:应用自制的AGEs(AGE-BSA)刺激NRK52E细胞,采用免疫细胞化学方法检测pm ad2/3核表达情况;ELISA方法检测细胞培养上清TGF-β1的浓度;RT-PCR检测TGF-β1、Sm ad2、Sm ad3和Sm ad7 mRNA的表达;W estern印迹检测α-平滑肌肌动蛋白(SMA)、E-钙粘着糖蛋白(cadherin)和1型胶原(collagenⅠ)蛋白的表达。结果:AGE-BSA刺激15 m in后pSm ad2/3核表达明显增加,于30 m in(68%)和24 h(76%)出现两个高峰,与刺激前及时间匹配的BSA对照组比较均有显著差异(P<0.05);AGE-BSA以时间依赖方式上调TGF-β1、Sm ad2、Sm ad3和Sm ad7 mRNA的表达;NRK52E细胞α-SMA和collagenⅠ蛋白表达高于对照组(P<0.01),E-cadherin蛋白表达低于对照组(P<0.01),细胞上清液TGF-β1的浓度高于对照组(P<0.01)。结论:AGEs可诱导肾小管上皮细胞Sm ads信号通路活化,促进肾小管上皮细胞转分化和细胞外基质collagenⅠ的合成。
AIM: To observe the effect of AGEs on the transdifferentiation, colla-genⅠ synthesis and Sm ads pathway in normal rat proximal renal tubular epithelial cells (NRK52E). Methods: NRK52E cells were stimulated with AGEs (AGE-BSA) and the expression of pm ad2 / 3 was detected by immunocytochemistry. The concentration of TGF-β1 was detected by ELISA. The expressions of TGF- Sm ad2, Sm ad3 and Sm ad7 mRNA expression; Western blot was used to detect the expression of α-SMA, E-cadherin and collagenⅠprotein. Results: The nuclear expression of pSm ad2 / 3 was significantly increased after AGE-BSA stimulation for 15 mins. There were two peaks at 30 min (68%) and 24 h (76%), which were compared with BSA control group (P <0.05). AGE-BSA upregulated the expression of TGF-β1, Sm ad2, Sm ad3 and Sm ad7 mRNA in a time-dependent manner. The expressions of α-SMA and collagen Ⅰ in NRK52E cells were higher than those in control group 0.01). The expression of E-cadherin was lower than that of the control group (P <0.01). The concentration of TGF-β1 in the cell supernatant was higher than that of the control group (P <0.01). CONCLUSION: AGEs can induce the activation of Sm ads signaling pathway in renal tubular epithelial cells and promote the renal tubular epithelial cell transdifferentiation and the synthesis of extracellular matrix collagen Ⅰ.