晚期糖基化终产物对高糖诱导NRK-52E细胞的转分化作用

Effect of Advanced Glycation End-products on High Gluclose Induced Transdifferentiation in NRK-52E Cells

目的:探讨晚期糖基化终产物(AGEs)对高糖诱导大鼠肾小管导管上皮细胞株NRK-52E的转分化干预作用。方法:大鼠肾小管导管上皮细胞株设空白组(不给葡萄糖)、高糖组(25mmol/L葡萄糖)、AGEs干预组(100mg/LAGEs)、高糖AGEs干预组(100mg/LAGEs+25mmol/L葡萄糖),培养72h后,用Westernbolt检测转化因子β1(TGF-β1)、Ⅰ型胶原、金属蛋白酶2(MMP2)、α平滑肌肌动蛋白(α-SMA)及甲状旁腺素相关肽(PTHrP)在NRK-52E细胞的表达,活性氧(ROS)含量应用CM2H2DCFDA试剂盒检测。结果:高浓度葡萄糖上调TGF-β1、Ⅰ型胶原、MMP2、α-SMA以及PTHrP表达,AGEs显著上调高浓度葡萄糖状态下TGF-β1、Ⅰ型胶原、MMP2、α-SMA和PTHrP表达,提高ROS含量。结论:AGEs可以促进高浓度葡萄糖诱导肾小管导管上皮细胞转分化。

Objective：To study the intervention effect of advanced glycation end-products（AGEs）on hyperglycemia induced transdifferentiation in NRK-52E cells.Methods：Four groups of NRK-52E cells were established：blank control group（group CON,was given no glucose and no intervention）,high glucose group（group HG,given 25mmol/L of glucose）,AGEs treated group（group AGE,given 100 mg/L of AGEs）,and AGEs plus high glucose treated group（group HG-AGE,given 100 mg/L of AGEs plus 25 mmol/L of glucose）,and Western blot was carried out to determine the expression of transforming growth factor β1（TGF-β1）,typeⅠcollagen,metalloprotease 2（MMP2）,smooth muscle actin α（α-SMA）,and parathyroid hormone-related peptide（PTHrP）in these cells after 72 h of reaction.Active oxygen（ROS）contents were detected by using CM2H2DCFDA kit.Results：The expression of TGF-β1,typeⅠcollagen,MMP2,α-SMA and PTHrP were up-regulated by glucose of high concentration;AGEs up-regulated TGF-β1,typeⅠcollagen,MMP2,α-SMA,and PTHrP under high glucose condition,and increased ROS content.Conclusion：AGEs could increase transdifferentiation induced by hyperglycemia in NRK-52E cells.