3种姜黄色素单体抑制人内皮细胞作用的实验研究

Inhibition of human endothelial cell line by curcumin and its derivatives

目的 探讨姜黄素、脱甲氧基姜黄素和双脱甲氧基姜黄素等 3种姜黄色素单体对人血管内皮细胞生长、增殖的抑制作用及其可能的作用机理。方法 (1)采用噻唑兰还原实验 (MTT)检测系列药物浓度和不同作用时间对两种血管内皮细胞增殖活性的影响 ;(2 )采用流式细胞术 (FCM)检测双脱甲氧基姜黄素作用 2 4h后内皮细胞的增殖周期改变。结果 (1)MTT实验表明 ,3种姜黄色素作用 72h后对内皮细胞增殖抑制作用的IC5 0值分别为 0 .5 2 5 μg/ml、0 .399μg/ml和 0 .12 5 μg/ml;在浓度为 4 μg/ml时 ,3种姜黄色素作用 4 8h后均导致内皮细胞出现明显的增殖抑制 (P <0 .0 5 ) ,作用 72h时达最高值 ;(2 )FCM检测结果表明 ,浓度为 4 μg/ml时双脱甲氧基姜黄素可将内皮细胞阻止于S期 (P <0 .0 5 ) ;浓度增至 8μg/ml以上时不但可引起S期细胞阻滞 (P<0 .0 1) ,同时又有明确的诱导凋亡作用。结论 (1) 3种姜黄色素均可明显抑制人内皮细胞的分裂与增殖 ;三者的生物学活性相差显著 :双脱甲氧基姜黄素 >脱甲氧基姜黄素 >姜黄素 (P <0 .0 5 ) ;(2 )影响细胞增殖周期和诱导凋亡是姜黄色素抑制内皮细胞增殖的主要作用机理之一。

Objective To investigate the inhibition of two endothelial cell lines by curcumin and its derivatives. Methods (1) MTT reduction assay was used to observe the different of the inhibition of two endothelial cell lines by curcumin or its derivatives; (2) Flow cytometry (FCM) was used to investigate if and how bisdemethoxycurcumin in different concentrations effects the cell cycle of endothelial cell in vitro after 24 hours of action. Results (1) MTT reduction assay indicated that the IC50 values of curcumin, demethoxycurcumin and bisdemethoxycurcumin are 0.525μg/ml?0.399μg/ml and 0.125μg/ml respectively when they were used to inhibit endothelial cell in vitro for 72 hours; The growth lines indicated that inhibitive effects of curcumin and its derivatives on endothelial cell were obvious after 48 hours and arrived at the top value. (2) FCM showed that bisdemethoxycurcumin of 4μg/ml retarded effectively endothelial cell line in vitro at S phase and apoptosis of endothelial cell line was induced when acted by bisdemethoxycurcumin of 8μg/ml or 12μg/ml for 24 hours. Conclusion (1) The inhibitive effects of curcumin, demethoxycurcumin and bisdemethoxycurcumin on endothelial cell in vitro are different: bisdemethoxycurcumin>demethoxycurcumin>curcumin; (2) Maybe effecting cell cycle and inducing cell apoptosis are part of mechanism of inhibitive effect of curcumin or its derivatives on endothelial cell in vitro.