摘要
目的探讨蔗糖铁刺激对肾小管上皮细胞纤维化的影响及其机制。方法使用0.1 mmol/L的蔗糖铁分别刺激HK-2细胞12 h、24 h、36 h,亚铁离子探针FeRhoNox及羟自由基探针(hydroxyphenyl fluorescein,HPF)检测HK-2细胞内亚铁及羟自由基含量,Western blot法检测细胞TGF-β1、CollagenⅠ、CollagenⅣ、Fibronectin蛋白表达。结果荧光半定量分析显示,0.1 mmol/L蔗糖铁刺激HK-2细胞12 h后,细胞内亚铁离子是对照组的(8.7±1.2)倍(P <0.01),细胞内羟基是对照组(9.0±1.0)倍(P <0.01);24 h后细胞内亚铁离子是对照组的(8.6±1.2)倍(P <0.01),细胞内羟基是对照组的(9.2±0.5)倍(P <0.01);36 h后细胞内亚铁离子是对照组的(8.8±0.8)倍(P <0.01),细胞内羟基是对照组的(8.6±1.0)倍(P <0.01)。0.1mmol/L蔗糖铁刺激HK-2细胞24 h和36 h后,TGF-β1、CollagenⅠ、CollagenⅣ、Fibronectin的表达明显增加。结论 0.1mmol/L的蔗糖铁刺激HK-2细胞增加细胞内氧化应激,与细胞纤维化相关。
Objective To investigate the effect of iron sucrose stimulation on renal tubular epithelial cell fibrosis and its mechanism.Methods HK-2 cells were stimulated with 0.1 mmol/L sucrose iron for 12 h, 24 h, 36 h. Ferrous ion probe FeRhoNox and hydroxyl radical probe hydroxyphenyl fluorescein(HPF) were used to detect ferrous and hydroxyl radicals in HK-2 cells. The expression of TGF-β1, collagenⅠ, collagenⅣand fibronectin were detected by western blotting. Results After stimulation of HK-2 cells with0.1 mmol/L sucrose iron for 12 h, 24 h and 36 h, the intracellular ferrous ions and the intracellular hydroxyl increased(all P<0.01).And the expression of TGF-β1, collagenⅠ, collagenⅣ, fibronectin also increased significantly. Conclusion After the stimulation of sucrose iron(0.1 mmol/L), intracellular ferrous and hydroxyl radicals increase, which is associated with cell fibrosis.
引文
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