Impact of Arachidonic Acid and the Leukotriene Signaling Pathway on Vasculogenesis of Mouse Embryonic Stem Cells

Abstract

Embryonic stem (ES) cells can differentiate into various kinds of cells, such as endothelial and hematopoietic cells. In addition, some evidence suggests that inflammatory mediators such as leukotrienes (LTs), which include the 5-lipoxygenase (LOX) family, can regulate endothelial cell differentiation. In the present study, the eicosanoid precursor arachidonic acid (AA) stimulated vasculogenesis of ES cells by increasing the number of fetal liver kinase-1+ vascular progenitor cells as well as vascular structures positive for platelet endothelial cell adhesion protein-1 and vascular endothelial cadherin. The stimulation of vasculogenesis and expression of the rate-limiting enzyme in the LT signaling pathway, 5-LOX-activating protein (FLAP), was blunted upon treatment with the FLAP inhibitors AM643 and REV5901. Vasculogenesis was significantly restored upon exogenous addition of LTs. Downstream of FLAP, the LTB₄ receptor (BLT1) blocker U75302, the BLT2 receptor blocker LY255283 as well as the cysteinyl LT blocker BAY-u9773 inhibited vasculogenesis of ES cells. AA treatment of differentiating ES cells increased reactive oxygen species (ROS) generation, which was not affected upon either FLAP or cyclooxygenase-2 inhibition. Prevention of ROS generation by either the free radical scavengers vitamin E and N-(2-mercaptopropionyl)glycine or the NADPH oxidase inhibitor VAS2870 downregulated vasculogenesis of ES cells and blunted the provasculogenic effect of AA. In summary, our data demonstrate that proinflammatory AA stimulates vasculogenesis of ES cells via the LT pathway by mechanisms involving ROS generation.

Omega‐3 and Omega‐6 polyunsaturated fatty acids stimulate vascular differentiation of mouse embryonic stem cells

“…Moreover, PUFA-stimulated vasculogenesis was blunted upon inhibition of ROS and NO generation by using pharmacologic NADPH oxidase and eNOS inhibitors. These data corroborate previous studies of us showing that the stimulation of vasculogenesis in ES cells by arachidonic acid was dependent on ROS generation (Huang, Sharifpanah, Becker, Wartenberg, & Sauer, 2016). It is well reported that upon chemical reaction of ROS with NO, peroxynitrite is formed which-in high concentrations-is generally regarded as very reactive and toxic substance and causes endothelial dysfunction (Kanaan & Harper, 2017).…”

A new treatment for severe pulmonary arterial hypertension based on an old idea: inhibition of 5‐lipoxygenase

“…19,20 Over the last four decades, the recognized biological actions of LTs have expanded well beyond smooth muscle contraction and chemotaxis, and a sampling of these can be found in references. [21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37] For example, 5-HETE and cysLTs are now recognized to stimulate cell proliferation and have been implicated in the development of colon and prostate cancer; 36,37 LTC 4 appears to be a major trigger of stressinduced oxidative damage. 21 The expression of LT biosynthetic enzymes as well as LT receptors are under the control of transcriptional and epigenetic mechanisms, in particular DNA methylation, and are themselves modulated by a variety of cytokines, growth factors like TGF beta, hormones, and inflammatory mediators.…”

“…Over the last four decades, the recognized biological actions of LTs have expanded well beyond smooth muscle contraction and chemotaxis, and a sampling of these can be found in references 21–37 . For example, 5‐HETE and cysLTs are now recognized to stimulate cell proliferation and have been implicated in the development of colon and prostate cancer; 36 , 37 LTC 4 appears to be a major trigger of stress‐induced oxidative damage 21 …”

Generation and phenotype analysis of CysLTR1 L118F mutant mice

“…It has been shown in recent reports that CysLTR1 signaling also modulates changes in portal pressure and liver tissue damage associated with acute cholestasis, as well as hepatic stellate cell activation, showing the critical roles of this receptor in the regulation of liver function . CysLTR1‐related signaling can also regulate embryonic stem cell differentiation, possibly mediated by signaling networks involving signal transducer and activator of transcription‐3, the phosphorylation of glycogen synthase kinase‐3β, and the generation of reactive oxygen species . The extensive involvement of CysLTR1‐mediated signaling in various cell types, particularly in stem cells, suggests that CysLTR1 might also regulate developmental processes, which merits further investigations.…”

“…21,22 CysLTR1-related signaling can also regulate embryonic stem cell differentiation, possibly mediated by signaling networks involving signal transducer and activator of transcription-3, the phosphorylation of glycogen synthase kinase-3β, and the generation of reactive oxygen species. 23,24 The extensive involvement of CysLTR1-mediated signaling in various cell types, particularly in stem cells, suggests that CysLTR1 might also regulate developmental processes, which merits further investigations.…”