Curcumin induces differentiation of embryonic stem cells through possible modulation of nitric oxide-cyclic GMP pathway
Curcumin induces differentiation of embryonic stem cells through possible modulation of nitric oxide-cyclic GMP pathway
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DOI：
10.1007/s13238-012-2053-2
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发表时间：
2012-07-01
期刊：
影响因子：
21.1
通讯作者：
Murad, Ferid
中科院分区：
文献类型：
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作者：
Mujoo, Kalpana;Nikonoff, Lubov E.;Murad, Ferid
姜黄素是咖喱中使用的膳食香料的活性成分，已被证明具有抗氧化、抗炎和抗增殖的特性。采用EB定向诱导H-9人胚胎干细胞分化的方法，观察了姜黄素（0-20 μ mol/L）对H-9人胚胎干细胞分化的促进作用。我们的结果使用真实的时间PCR，蛋白质印迹和免疫染色表明，姜黄素显着增加心脏特异性转录因子NKx 2.5，心肌肌钙蛋白I，肌球蛋白重链，和内皮型一氧化氮合酶的基因表达和蛋白水平在ES细胞分化。此外，NO供体增强了姜黄素介导的NKx2.5和其他心脏特异性蛋白的诱导。用姜黄素孵育细胞会导致细胞内亚硝酸盐的剂量依赖性增加，其程度与提供真正的NO供体相同。第二信使环腺苷酸（cAMP）和环鸟苷酸（cGMP）的功能测定显示，姜黄素在分化细胞中的持续存在诱导cAMP的基线水平降低，但其显著升高cGMP的基线含量。姜黄素除了无细胞测定显着抑制cAMP和cGMP降解的提取物，而长期治疗的完整细胞与姜黄素增加cAMP和cGMP降解的速率，表明这可能是由于直接抑制一些环核苷酸降解酶（磷酸二酯酶）的姜黄素。这些研究表明，多酚姜黄素可能参与ES细胞的分化，部分是由于操纵一氧化氮信号。
Curcumin, an active ingredient of dietary spice used in curry, has been shown to exhibit anti-oxidant, anti-inflammatory and anti-proliferative properties. Using EB directed differentiation protocol of H-9 human embryonic stem (ES) cells; we evaluated the effect of curcumin (0-20 mu mol/L) in enhancing such differentiation. Our results using real time PCR, western blotting and immunostaining demonstrated that curcumin significantly increased the gene expression and protein levels of cardiac specific transcription factor NKx2.5, cardiac troponin I, myosin heavy chain, and endothelial nitric oxide synthase during ES cell differentiation. Furthermore, an NO donor enhanced the curcumin-mediated induction of NKx2.5 and other cardiac specific proteins. Incubation of cells with curcumin led to a dose dependent increase in intracellular nitrite to the same extent as giving an authentic NO donor. Functional assay for second messenger(s) cyclic AMP (cAMP) and cyclic GMP (cGMP) revealed that continuous presence of curcumin in differentiated cells induced a decrease in the baseline levels of cAMP but it significantly elevated baseline contents of cGMP. Curcumin addition to a cell free assay significantly suppressed cAMP and cGMP degradation in the extracts while long term treatment of intact cells with curcumin increased the rates of cAMP and cGMP degradation suggesting that this might be due to direct suppression of some cyclic nucleotide-degrading enzyme (phosphodiesterase) by curcumin. These studies demonstrate that polyphenol curcumin may be involved in differentiation of ES cells partly due to manipulation of nitric oxide signaling.