间歇性高糖对NRK-52E细胞抗氧化能力作用研究

Effect of fluctuating high glucose on inhibition of anti-oxidative stress in NRK-52E cell line

目的探讨间歇性高糖抑制大鼠肾小管导管上皮细胞株NRK-52E抗氧化能力。方法实验开始72 h后,Western blot检测空白组、正常葡萄糖组(5 mmol/L葡萄糖)、高糖干预组(25 nmol/L葡萄糖)、间歇性高糖干预组(5 mmol/L葡萄糖与25 mmol/L葡萄糖交替)中氧化物歧化酶-1(SOD-1)、还原型辅酶Ⅱ(NADPH)在NRK-52E细胞表达。一氧化氮合酶(eNOS)试剂盒应用液闪仪测定3[H]的放射强度,活性氧(ROS)含量应用CM2H2DCFDA试剂盒检测。结果高浓度葡萄糖及间歇性高糖均下调SOD-1及NADPH表达,而提高ROS含量,间歇性高糖作用更显著。间歇性高糖eNOS含量低于持续性高糖组。结论间歇性高糖抑制肾小管导管上皮细胞抗氧化能力显著。

Objective To study whether fluctuating high glucose had inhibited anti- oxidative stress in NRK-52E cell line. Methods Western blot in control group（0 mmol/L, glucose）, normal glucose group（5 mmol/L glucose）, high glucose group （25 mmol/L glucose）, and fluctuating high glucose group （5 mmol/L glucose/25 mmol/L glucose） had carried out. Then NOS assay kit was used to measure the activity of eNOS, ROS were detected by CM2H2DCFDA after 72h. Results The expression of SOD-1 and NADPH were down-regulated by hyperglycemia and fluctuating high glu- cose. Fluctuating high glucose also could increase more ROS than high glucose. The level of eNOS in fluctuating high glucose was lower than that of high glucose. Conclusion Fluctuating high glucose could inhibit anti-oxidative stress in NRK-52E cell line.