Induction of cytotoxicity and apoptosis and inhibition of cyclooxygenase-2 gene expression, by curcumin and its analog, alpha-diisoeugenol.
Induction of cytotoxicity and apoptosis and inhibition of cyclooxygenase-2 gene expression, by curcumin and its analog, alpha-diisoeugenol.
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发表时间：
2005-11
影响因子：
2
通讯作者：
T. Atsumi;Y. Murakami;K. Shibuya;K. Tonosaki;S. Fujisawa
中科院分区：
文献类型：
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作者：
T. Atsumi;Y. Murakami;K. Shibuya;K. Tonosaki;S. Fujisawa
研究了细胞毒性和α-二异丁香酚。姜黄素和α-二异丁香酚对人早幼粒细胞白血病细胞（HL-60细胞）和人下颌下癌细胞（HSG细胞）的细胞毒性相似（CC 50 1-3 μ M）。然而，姜黄素诱导更多的凋亡，特别是在HL-60细胞与HSG细胞相比，通过测量亚G1/G 0 DNA分数在流式细胞术直方图中显示。用15 μ M姜黄素处理使HL-60细胞中具有亚G1/G 0 DNA部分的细胞数量从对照水平<5%增加到55%，在HSG细胞中增加到30%。流式细胞术，在用annexin V-FITC/PI染色后（暴露于凋亡细胞表面的磷脂酰丝氨酸（PS）），显示姜黄素诱导早期凋亡的剂量依赖性，在用10 μ M姜黄素处理后，HL-60细胞中达到约65%，在HSG细胞中达到约20%。相比之下，α-二异丁香酚未能诱导任何细胞类型的凋亡。对于两种细胞类型，在姜黄素和α-二异丁香酚浓度大于10 μ M时，晚期凋亡/坏死细胞的比例迅速增加。CDFH-DA染色显示姜黄素处理的HL-60细胞内活性氧（ROS）的产生量高于HSG细胞。在两种细胞类型中，由α-二异丁香酚产生的ROS处于对照水平。姜黄素产生的ROS被抑制的抗氧化剂，如N-乙酰-L-半胱氨酸（NAC）和谷胱甘肽（GSH）和羟基自由基的清除剂，如甘露醇，但相反，促进促氧化剂，如过渡金属离子Cu（II）和Zn（II）。ROS的产生可能在PS暴露中起一定作用。在RAW 264.7细胞中，10 μ M的姜黄素抑制LPS（脂多糖）诱导的考克斯-2基因表达，但不抑制α-二异丁香酚。半经验PM 3计算表明，姜黄素的这种活性，其中它表现为非甾体抗炎药（NSAID）样化合物，取决于其酚功能，这比α-二异丁香酚更明显。总之，我们的研究结果表明，姜黄素的生物活性是其作为促氧化剂和抗氧化剂的能力的结果。
Cytotoxici and alpha-diisoeugenol were investigated. The cytotoxicity of curcumin and a-diisoeugenol against human promyelocytic leukemia cells (HL-60 cells) and human submandibular cancer cells (HSG cells) was similar (CC50 1-3 microM). However, curcumin induced much more apoptosis, particularly in HL-60 cells compared with HSG cells, as revealed by measurement of the sub-G1/G0 DNA fraction in flow cytometric histograms. Treatment with 15 microM curcumin increased the number of cells with a sub-G1/G0 DNA fraction from control levels of <5% to 55% in HL-60 cells and 30% in HSG cells. Flow cytometry, after staining with annexin V-FITC/PI (the exposure of phosphatidylserine (PS) on the surface of apoptotic cells), showed a dose-dependent induction of early apoptosis by curcumin, which reached about 65% in HL-60 cells and about 20% in HSG cells after treatment with 10 microM curcumin. In contrast, alpha-diisoeugenol failed to induce apoptosis in either cell type. For both cell types, the proportion of late apoptotic/necrotic cells increased rapidly at concentrations of curcumin and a-diisoeugenol greater than 10 microM. The generation of intracellular reactive oxygen species (ROS) in curcumin-treated HL-60 cells was greater than that in HSG cells, as judged by CDFH-DA staining. In both cell types, ROS generation by a-diisoeugenol was at control levels. ROS generation by curcumin was suppressed by antioxidants such as N-acetyl-L-cysteine (NAC) and glutathione (GSH) and by scavengers of hydroxy radicals such as mannitol, but, conversely, was promoted by prooxidants such as the transition metal ions Cu(II) and Zn(II). ROS generation may play a part in the exposure of PS. Curcumin, but not a-diisoeugenol, at 10 microM inhibited LPS (lipopolysaccharide)-induced COX-2 gene expression in RAW 264.7 cells. Semiempirical PM 3 calculations suggested that this activity of curcumin, in which it behaves as a non-steroidal anti-inflammatory drug (NSAID)-like compound, is dependent on its phenolic function, which is more pronounced than that of alpha-diisoeugenol. Taken together, our results suggest that the bioactivity of curcumin is a result of its ability to act as both a prooxidant and an antioxidant.