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微摩尔浓度铜离子通过刺激活性氧产生促进人角质形成细胞增殖

, PP. 1053-1058

Keywords: 铜离子,人角质形成细胞,细胞增殖,活性氧,N-乙酰半胱氨酸

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Abstract:

目的观察不同浓度Cu2+对人角质形成细胞株(HaCaT)增殖活性及细胞内活性氧(reactiveoxygenspecies,ROS)水平的影响,用活性氧清除剂N-乙酰半胱氨酸(NAC)验证两者之间的关系。方法不同浓度Cu2+(0、10-8、10-7、10-6、10-5、10-4、10-3mol/L)组或(和)5mmol/LNAC处理HaCaT细胞后,采用CCK-8法和DCFH-DA分别检测细胞的增殖活性和细胞内ROS产生,共聚焦显微镜观察细胞内ROS产生及其与线粒体的分布情况。结果与阴性对照组(0mol/LCu2+)相比,10-6、10-5mol/L组Cu2+处理48h后HaCaT细胞相对增殖率分别升高至116.19%(P=0.027)、113.37%(P=0.036),而其细胞内ROS产生在处理后30min达到127.47%(P=0.001)、124.28%(P=0.031),60min进一步上升至140.47%(P=0.002)、134.29%(P=0.007)。5mmol/LNAC能拮抗微摩尔浓度Cu2+的上述作用。虽然10-8、10-7mol/LCu2+在处理后60min也刺激细胞内ROS产生明显升高至110.99%(P=0.016)及117.34%(P=0.016),但是处理48h后细胞相对增殖率仅为101.09%和101.86%,并无促进细胞增殖的效应(P>0.05)。结论微摩尔浓度Cu2+能促进人角质形成细胞增殖,这种促增殖效应可能与刺激细胞内ROS水平的适度上调有关。

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