姜黄素衍生物对K562细胞酪氨酸激酶活性的影响

目的研究姜黄素（Cur）衍生物对K562细胞酪氨酸酶激活性的影响,从中筛选酪氨酸激酶（PTKs）抑制剂。方法以PGT聚谷氨酸钠∶酪氨酸（4∶1）]为激酶反应底物包被96孔酶标板,以磷酸化酪氨酸特异性单克隆抗体为第一抗体,应用酶联免疫吸附测定法（ELISA）检测反应体系中加入的K562细胞裂解液PTKs对反应底物PGT的酪氨酸残基磷酸化的程度,以此测定K562细胞裂解液中的PTKs活性。在此反应体系中加入不同浓度的Cur衍生物,观察对PTKs活性的影响。结果对Cur衍生物进行筛选,得到4种PTKs抑制剂,分别为Cur衍生物FM0815、FM0817、FM0822及FM0824。4个Cur衍生物对PTKs活性的抑制均明显强于Cur,且呈量效、时效关系。结论 Cur衍生物FM0815、FM0817、FM0822及FM0824具有显著的PTKs活性抑制作用。

Effect of Curcumin Derivatives on Activity of Protein Tyrosine Kinase in K562 cells

Objective To study the effect of curcumin derivatives on activity of protein tyrosine kinase（PTK） in K562 cells and to screen potential PTK inhibitors.Methods Poly Glu∶Tyr（4∶1） coated on the 96-well ELISA plate was used as the substrate and phosphotyrosine-specific monoclonal antibody as the first antibody to determine the level of PGT tyrosine residue phosphorylation which reflected the activity of protein tyrosine kinase in K562 cell lysate.Curcumin derivatives of different concentrations were screened and their effects on the activity of PTK were studied.Results Curcumin derivatives were screened and 4 potential inhibitors were obtained,namely,FM0815,FM0817,FM0822 and FM0824.All of them had much higher PTK inhibition activity than curcumin in a dose and time-dependent manner.Conclusion The activity of PTK is inhibited apparently by curcumin derivatives FM0815,FM0817,FM0822 and FM0824.