A profile of NSAID-targeted arachidonic acid metabolisms in human embryonic stem cells (hESCs): implication of the negative effects of NSAIDs on heart tissue regeneration.
A profile of NSAID-targeted arachidonic acid metabolisms in human embryonic stem cells (hESCs): implication of the negative effects of NSAIDs on heart tissue regeneration.
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DOI：
10.1016/j.ijcard.2010.04.015
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发表时间：
2011-08
影响因子：
3.5
通讯作者：
A. Chillar;Shuiping So;Cheng-Huai Ruan;H. Shelat;Y. Geng;K. Ruan
中科院分区：
文献类型：
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作者：
A. Chillar;Shuiping So;Cheng-Huai Ruan;H. Shelat;Y. Geng;K. Ruan
引言利用人胚胎干细胞（hESCs）再生梗死心脏组织的新兴技术一直不发达。然而，由于非甾体抗炎药（NSAIDs），如阿司匹林，在梗死期间，它变得至关重要，知道是否有负面影响的NSAIDs对心脏组织再生时，使用hESCs. METHODS质谱（LC/MS/MS）和高效液相色谱（HPLC）分析用于分析功能存在的精心制作的前列腺素类化合物的生物合成和信号系统在hESCs。检测到的hESC膜中释放的内源性花生四烯酸（AA）反映了磷脂酶的活性，磷脂酶直接控制前列腺素类的生物合成。环加氧酶-2（考克斯-2）抑制剂NS 398对内源性前列腺素E2（PGE 2）生物合成的完全抑制证实了hESC中合成的主要前列腺素类是由考克斯-2酶介导的。结论不同的环氧合酶（考克斯）下游酶和代谢酶通过考克斯-1和考克斯-2途径参与AA产物的信号转导。通过蛋白质印迹证实了参与前列腺素类生物合成和活性的许多酶和受体[（考克斯-1、考克斯-2、微粒体前列腺素E合酶（mPGES）、胞质前列腺素E合酶（cPGES）、前列腺素I合酶（PGIS）、PGE 2亚型受体（EP 1、EP 2和EP 4）和前列环素受体（IP）]的存在。这些研究暗示了NSAID的负面作用，例如阿司匹林和考克斯-2抑制剂，当使用hESC时，它们在梗死心脏的组织再生期间抑制前列腺素类的产生。
INTRODUCTIONAn emerging technology using human embryonic stem cells (hESCs) to regenerate infarcted heart tissue has been underdeveloped. However, because non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin, are taken during the infarction, it becomes critical to know whether the NSAIDs have negative impacts on heart tissue regeneration when using hESCs.METHODSMass spectrometry (LC/MS/MS) and high performance liquid chromatography (HPLC) analyses were used to analyze the functional presence of the elaborate prostanoids' biosynthesis and signaling systems in hESCs. The detected endogenous arachidonic acid (AA) released in the hESC membranes reflects the activity of phospholipase which directly controls the biosyntheses of the prostanoids.RESULTSThe complete inhibition of the endogenous prostaglandin E2(PGE2) biosynthesis by the cyclooxygenase-2 (COX-2) inhibitor, NS398, confirmed that the major prostanoids synthesized in the hESCs are mediated by the COX-2 enzyme. We also found that PGE2and the prostacyclin (PGI2) metabolite, 6-keto-PGF1α, are present in the undifferentiated hESCs.CONCLUSIONThis indicated different cyclooxygenase (COX)-downstream synthases and metabolizing enzymes are involved in the AA products' signaling through the COX-1 and COX-2 pathways. The presence of many enzymes' and receptors' [(COX-1, COX-2, microsomal prostaglandin E synthase (mPGES), cytosolic prostaglandin E synthase (cPGES), prostaglandin I synthase (PGIS), the PGE2subtype receptors (EP1, EP2, and EP4) and the prostacyclin receptor (IP)] involvement in the prostanoid biosynthesis and activity was confirmed by western blot. The studies implied the negative effects of NSAIDs, such as aspirin and COX-2 inhibitors, which suppress prostanoid production during tissue regeneration for infarcted heart when using hESCs.