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目的:通过观察尿酸对肾小管上皮细胞赖氨酸氧化酶表达的影响,探讨尿酸在肾小管损伤中的作用及其可能的机制。方法:以正常大鼠肾小管上皮细胞株NRK-52E细胞为研究对象,以不同浓度的尿酸作用于该细胞,采用免疫蛋白印迹和间接免疫荧光的方法检测赖氨酸氧化酶(LOX)的表达水平,以及细胞外基质蛋白纤维连接蛋白(fibronectin,FN)表达的变化。观察用小干扰RNA下调LOX对尿酸诱导FN表达的影响。结果:尿酸能够显著上调NRK-52E细胞LOX前体及成熟蛋白的合成,并呈现明显的剂量依赖关系;同时,FN的表达亦显著增加。LOX siRNA能够显著抑制尿酸诱导NRK-52E细胞表达成熟LOX的作用;同时LOX siRNA能够降低高尿酸导致的FN表达的增加。结论:尿酸能够通过上调赖氨酸氧化酶增加FN的表达。
Objective: To investigate the effect of uric acid on the expression of lysine oxidase in renal tubular epithelial cells and explore its possible mechanism. Methods: Normal rat renal tubular epithelial cell line NRK-52E cells were treated with different concentrations of uric acid, and the expression of lysine oxidase (LOX) was detected by Western blotting and indirect immunofluorescence Level, and the expression of extracellular matrix fibronectin (FN). The effect of down-regulating LOX on uric acid-induced FN expression with small interfering RNA was observed. RESULTS: Uric acid significantly increased the LOX precursor and mature protein synthesis in NRK-52E cells in a dose-dependent manner. At the same time, the expression of FN was also significantly increased. LOX siRNA can significantly inhibit uric acid-induced mature LOX expression in NRK-52E cells; while LOX siRNA can reduce hyperuricemia-induced increase in FN expression. Conclusion: Uric acid can increase the expression of FN by up-regulating lysine oxidase.