血管紧张素Ⅱ对高糖诱导NRK-52E细胞转分化作用研究

Effect of angiotensin Ⅱ on hyperglycemia-induced transdifferentiation in NRK-52E cells

目的探讨血管紧张素Ⅱ(AngⅡ)对高糖诱导大鼠肾小管导管上皮细胞株NRK-52E转分化的干预作用。方法细胞培养72 h后,Western blot检测对照组(0 mmol/L葡萄糖)、高糖组(25 mmol/L葡萄糖)、血管紧张素Ⅱ干预组(10 nmol/L AngⅡ)、高糖血管紧张素Ⅱ干预组(10 nmol/L AngⅡ+25 mmol/L葡萄糖)中转化生长因子β1(TGF-β1)、Ⅰ型胶原、金属蛋白酶2(MMP2)、α平滑肌肌动蛋白(α-SMA)以及甲状旁腺素相关肽(PTHrP)在NRK-52E细胞中的表达。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。AngⅡ明显上调高浓度葡萄糖状态下TGF-β1、Ⅰ型胶原、MMP2、α-SMA和PTHrP的表达,并提高ROS含量。结论AngⅡ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 investigate the effect with angiotensin Ⅱ intervention on hyperglycemia-induced transdifferentiation in the epithelioid clone of mixed culture of normal rat kidney（NRK-52E） cells.Methods 72 hours after intervention,the expression of transforming growth factor-β1（TGF-β1）,type Ⅰ collagen,matrix metallopeptidase 2（MMP2）,α-smooth muscle actin（α-SMA） and parathyroid hormone-related protein（PTHrP） in NRK-52E cells was detected with Western blot in the control group（0 mmol/L glucose）,high glucose group（25 mmol/L glucose）,angiotensin Ⅱ（10 nmol/L angiotensin Ⅱ） group and angiotensin Ⅱ plus high glucose（10 nmol/L angiotensin Ⅱ＋25mmol/L glucose） group,respectively;and the levels of reactive oxygen species（ROS） were detected by kit CM2H2DCFDA.Results The expression of TGF-β1,type Ⅰ collagen,MMP2,α-SMA and PTHrP were up-regulated by hyperglycemia;angiotensin Ⅱ markedly up-regulated TGF-β1,type Ⅰ collagen,MMP2,α-SMA and PTHrP in the high glucose condition and increased ROS levels.Conclusion Angiotensin Ⅱ could promote the hyperglycemia-induced transdifferentiation in NRK-52E cells.