Lesinurad调节AMPK/mTOR抑制自噬改善高尿酸诱导的血管平滑肌异常增殖

Lesinurad improves high uric acid⁃induced abnormal proliferation of vascular smooth muscle by inhibiting autophagy via regulating AMPK/mTOR pathway

目的:探索Lesinurad改善高尿酸诱导的血管平滑肌异常增殖的作用及机制。方法:通过CCK⁃8法评价Lesinurad对高尿酸诱导的大鼠血管平滑肌细胞(A10细胞)增殖的保护作用;Western印迹检测Lesinurad对高尿酸诱导的A10细胞对尿酸转运体URAT1、自噬相关蛋白(LC3、Beclin1、p62)与AMPK/mTOR通路的影响。结果:900μmol/L尿酸可引起A10细胞异常增殖,25μmol/L Lesinurad均可明显抑制A10的异常增殖,降低其细胞活力。进一步研究发现,25μmol/L Lesinurad可明显尿酸转运体URAT1的表达,抑制高尿酸诱导的A10细胞自噬,显著下调LC3与Beclin1蛋白表达并上调p62蛋白表达水平。同时,25μmol/L Lesinurad显著抑制AMPK/mTOR通路的磷酸化过程。结论:Lesinurad能抑制高尿酸诱导的A10细胞增殖,其机制可能与调节AMPK/mTOR通路抑制自噬有关。

Objective:To investigate the effect of Lesinurad on improving the abnormal proliferation of vascular smooth muscle induced by high uric acid and its mechanism.Methods:The protective effect of Lesinurad against the proliferation of rat vascular smooth muscle cells(A10 cells)induced by high uric acid was evaluated by CCK⁃8 assay.Western blotting was performed to determine the effects of Lesinurad on the expression of urate transporter(URAT1),autophagy⁃related proteins(LC3,Beclin1,p62),and AMPK/mTOR pathway in A10 cells induced by high uric acid.Results:900μmol/L uric acid caused abnormal proliferation of A10 cells,and 25μmol/L Lesinurad significantly inhibited the abnormal proliferation of A10 cells and decreased the cell viability.Further studies found that 25μmol/L Lesinurad significantly inhibited the expression of URAT1 and autophagy in A10 cells induced by high uric acid,significantly down⁃regulated the protein expression of LC3 and Beclin1,and up⁃regulated the protein expression of p62.Meanwhile,25μmol/L Lesinurad significantly inhibited the phosphorylation of AMPK/mTOR pathway.Conclusion:Lesinurad may inhibit the proliferation of A10 cells induced by high uric acid,and its mechanism may be related to the inhibition of autophagy by regulating AMPK/mTOR pathway.