论文部分内容阅读
目的探讨氯沙坦对高糖及间歇性高糖诱导大鼠肾小管导管上皮细胞株NRK-52E转分化的抑制作用。方法实验分为空白组(0mmol/L葡萄糖)、正常葡萄糖组(5mmol/L葡萄糖)、高糖干预组(25mmol/L葡萄糖)、间歇性高糖干预组(5mmol/L葡萄糖/25mmol/L葡萄糖交替)、氯沙坦干预组(10μmol/L氯沙坦)、氯沙坦高糖干预组(10μmol/L氯沙坦预处理后再高糖培养)、氯沙坦间歇性高糖干预组(10μmol/L氯沙坦预处理后再间歇性高糖培养),根据分组对NRK-52E细胞施加相应因素作用72h后,蛋白免疫印迹法检测转分化标志物转化生长因子β1(TGF-β1)、Ⅰ型胶原、金属蛋白酶2(MMP-2)、α平滑肌肌动蛋白(α-SMA)以及甲状旁腺素相关肽(PTHrP)在细胞中的表达,CM2H2DCFDA试剂盒检测细胞ROS含量。结果高糖干预及间歇性高糖干预组TGF-β1、Ⅰ型胶原、MMP-2、α-SMA和PTHrP表达上调,ROS含量明显上升,间歇性高糖作用更显著。应用氯沙坦干预后可以部分下调高糖或间歇性高糖诱导的TGF-β1、Ⅰ型胶原、MMP-2、α-SMA以及PTHrP表达,降低细胞ROS含量。结论氯沙坦可抑制高糖及间歇性高糖诱导的NRK-52E细胞转分化作用。
Objective To investigate the inhibitory effect of losartan on the transdifferentiation of rat renal tubular epithelial cell line NRK-52E induced by high glucose and intermittent high glucose. Methods The experiment was divided into blank group (0mmol / L glucose), normal glucose group (5mmol / L glucose), high glucose intervention group (25mmol / L glucose), intermittent high glucose intervention group (5mmol / L glucose / 25mmol / L glucose Losartan intervention group (10μmol / L losartan), Losartan high glucose intervention group (10μmol / L losartan pretreatment and then high glucose culture), Losartan intermittent high glucose intervention group ( The cells were treated with 10μmol / L losartan for a period of high glucose, and then treated with NRK-52E cells for 72h. Western-blotting was used to detect the expression of transforming growth factor-β1 (TGF-β1) The expression of collagen type Ⅰ, MMP-2, α-SMA and PTHrP in cells were detected by MTT assay. The level of ROS was detected by CM2H2DCFDA kit. Results The expression of TGF-β1, type Ⅰ collagen, MMP-2, α-SMA and PTHrP were upregulated in high glucose intervention group and intermittent high glucose intervention group. ROS content increased obviously and intermittent hyperglycemia was more significant. Losartan intervention could partly down-regulate the expression of TGF-β1, type Ⅰ collagen, MMP-2, α-SMA and PTHrP induced by high glucose or intermittent high glucose, and reduce ROS content in cells. Conclusion Losartan can inhibit high glucose and intermittent high glucose-induced transdifferentiation of NRK-52E cells.