Synthesis and release of acetylcholine by cultured bovine arterial endothelial cells

Kawashima, Koichiro; Watanabe, Naomi; Oohata, Hisayo; Fujimoto, Kazuko; Suzuki, Takeshi; Ishizaki, Yasuki; Morita, Ikuo; Murota, Sei‐itsu

doi:10.1016/0304-3940(90)90822-q

Cited by 81 publications

(47 citation statements)

References 9 publications

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“…The endogenous ligand of the receptor is the nerve transmitter acetylcholine, which is abundant in human skin. Endothelial cells are also capable of making acetylcholine, 12,27 which may act in an autocrine or paracrine manner to stimulate endothelial cell proliferation and angiogenesis (unpublished observations from our group).…”

Section: Discussionmentioning

confidence: 88%

Nicotine Accelerates Angiogenesis and Wound Healing in Genetically Diabetic Mice

Jacobi

Jang

Sundram

et al. 2002

The American Journal of Pathology

157

117

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Recently, we have discovered an endogenous cholinergic pathway for angiogenesis mediated by endothelial nicotinic acetylcholine receptors (nAChRs). Since angiogenesis plays a major role in wound repair, we hypothesized that activation of nAChRs with nicotine would accelerate wound healing in a murine excisional wound model. In genetically diabetic and control mice full-thickness skin wounds (0.8 cm) were created on the dorsum and topically treated over 7 days with either vehicle (phosphate-buffered saline, PBS) or nicotine (10 ؊8 mol/L, 10 ؊9 mol/L; each, n ‫؍‬ 5). Wound size was measured over 14 days followed by resection, histological analysis, and quantitation of vascularity. In diabetic animals an agonist (epibatidine, 10 ؊10 mol/L) or antagonist (hexamethonium, 10 ؊4 mol/L) of nAChRs as well as the positive control basic fibroblast growth factor (bFGF, 25 g/kg) were also tested. To further study the role of endothelial nAChRs in angiogenesis, we used an ex vivo vascular explant model. In diabetic mice wound healing was markedly impaired. Nicotine significantly accelerated wound healing as assessed by closure rate and histological score. The effects of nicotine were equal to bFGF and were mimicked by epibatidine and blocked by hexamethonium. Histomorphometry revealed increased neovascularization in animals treated with nicotine. Furthermore, capillary-like sprouting from vascular explants was significantly enhanced by nicotine. In conclusion, agonist-induced stimulation of nAChRs accelerates wound healing in diabetic mice by promoting angiogenesis. We have discovered a cholinergic pathway for angiogenesis that is involved in wound healing, and which is a potential target for therapeutic angiogenesis.

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Section: Discussionmentioning

confidence: 88%

Nicotine Accelerates Angiogenesis and Wound Healing in Genetically Diabetic Mice

Jacobi

Jang

Sundram

et al. 2002

The American Journal of Pathology

157

117

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“…Alternatively, the artery may be the source of ACh. ECs contain choline acetyltransferase and synthesize and release ACh (29,36). However, the factors that regulate endothelial ACh release are not known, and a role for endogenous endothelial ACh has not been demonstrated.…”

Section: Discussionmentioning

confidence: 99%

Role of phospholipase C and diacylglyceride lipase pathway in arachidonic acid release and acetylcholine-induced vascular relaxation in rabbit aorta

Tang¹,

Edwards²,

Holmes³

et al. 2006

American Journal of Physiology-Heart and Circulatory Physiology

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(ECs). In rabbit aorta, AA is metabolized through the 15-lipoxygenase pathway to form vasodilatory eicosanoids 15-hydroxy-11,12-epoxyeicosatrienoic acid (HEETA) and 11,12,15-trihydroxyeicosatrienoic acid (THETA). AA is released from phosphatidylcholine (PC) and phosphatidylethanolamine (PE) by phospholipase A 2 (PLA2), or from phosphatidylinositol (PI) by phospholipase C (PLC) pathway. The diacylglycerol (DAG) lipase can convert DAG into 2-arachidonoylglycerol from which free AA can be released by monoacylglycerol (MAG) lipase or fatty acid amidohydrolase (FAAH). We used specific inhibitors to determine the involvement of the PLC pathway in ACh-induced AA release. In rabbit aortic rings precontracted by phenylephrine, ACh induced relaxation in the presence of indomethacin and N -nitro-L-arginine (L-NNA). These relaxations were blocked by the PLC inhibitor U-73122, DAG lipase inhibitor RHC-80267, and MAG lipase/FAAH inhibitor URB-532. Cultured rabbit aortic ECs were labeled with [ 14 C]AA and stimulated with methacholine (10 Ϫ5 M). Free [ 14 C]AA was released by methacholine. Methacholine decreased the [ 14 C]AA content of PI, DAG, and MAG fractions but not PC or PE fractions. Methacholine-induced release of [ 14 C]AA was blocked by U-73122, RHC-80267, and URB-532 but not by U-73343, an inactive analog of U-73122. The data suggested that ACh activates PLC, DAG lipase, and MAG lipase pathway to release AA from membrane lipids. This pathway is important in regulating vasodilatory eicosanoid synthesis and vascular relaxation in rabbit aorta. endothelium-dependent hyperpolarizing factor; phospholipid; monoacylglycerol lipase; trihydroxyeicosatrienoic acid; hydroxyepoxyeicosatrienoic acid VASCULAR ENDOTHELIAL CELLS (ECs) regulate vascular tone by synthesis and release of many bioactive compounds. Ligands such as hormones, ACh, bradykinin, and cytokines stimulate release of arachidonic acid (AA) from cellular membranes (6,15,26,43,55), and AA induces endothelium-dependent vasodilation in isolated rabbit aorta preconstricted with phenylephrine, which is not blocked by nitric oxide (NO) synthase (NOS) and cyclooxygenase (COX) inhibitors (19,39,50). The phospholipase inhibitors mepacrine and dimethyl-eicosadienoic acid (DEDA) decrease relaxation to ACh but not AA, suggesting AA or a metabolite may be involved in the relaxations (11). AA is metabolized by COX, lipoxygenase (LO), and cytochrome P-450 pathways into bioactive eicosanoids (10,30,40,48). Using specific inhibitors in rabbit aorta, Singer and Peach (51) first concluded that a LO metabolite mediates the NO-and PG-resistant relaxation to ACh (51). The LO inhibitors cinnamyl-3,4-dihydroxy-␣-cyanocinnamate, nordihydroguaiaretic acid, and ebselen block ACh and AA-induced vasodilation in rabbit aorta (11,39,42). Previous studies in our lab identified the 15-LO metabolites of AA in rabbit aorta. AA is oxidized by 15-LO to 15-hydroperoxyeicosatetraenoic acid (15-HPETE), which is converted to hydroxyepoxyeicosatrienoic acid (HEETA) and trihydroxyeicosatrienoic acid (THET...

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“…This represents a possible source of ligand for nAChRs present on bronchial epithelial cells (Wang et al, 2001). Vascular endothelial cells themselves synthesize acetylcholine suggesting a source of the ligand for autocrine activation of nAChRs (Kawashima et al, 1990). In keratinocytes, there is a spatial pattern of distribution of both choline acetyltransferase (ChAT) and AChE.…”

Section: Possible Sources Of Ach For Nachr Activation In Nonexcitablementioning

confidence: 99%

“…Although these studies have implicated nAChRs in atherosclerosis and in tumor promotion (by increasing tumor vascularization), it is not clear whether angiogenesis represents a physiologic function for nAChRs. As endothelial cells themselves can produce ACh (Kawashima et al, 1990), this could potentially be an autocrine mechanism for blood vessel production as well as regulation of their tonicity.…”

Section: Possible Physiologic Roles For Nachrs On Nonexcitable Cellsmentioning

confidence: 99%

Nicotinic receptor signaling in nonexcitable cells

2002

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ABSTRACT:The finding that neuronal nicotinic acetylcholine receptors (nAChRs) are present in nonneuronal cells both within and outside the nervous system raises some interesting issues. The mechanisms underlying receptor signaling and its downstream consequences in these cells remain to be elucidated. Factors controlling the release of acetylcholine and the extent of its diffusion are likely to be different for these cells than for traditional neuronal synapses. Recent advances on the physiologic functions of some of these cell types have provided a better insight into possible functional roles for nAChRs in nonexcitable cells. The presence of nAChRs on these cells also implies a broader scope for the actions of nicotine that needs to be considered from a clinical viewpoint. Revealing the potential physiologic roles for nAChRs on nonexcitable cells is likely to provide a more complete understanding of cholinergic signaling.

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Synthesis and release of acetylcholine by cultured bovine arterial endothelial cells

Cited by 81 publications

References 9 publications

Nicotine Accelerates Angiogenesis and Wound Healing in Genetically Diabetic Mice

Nicotine Accelerates Angiogenesis and Wound Healing in Genetically Diabetic Mice

Role of phospholipase C and diacylglyceride lipase pathway in arachidonic acid release and acetylcholine-induced vascular relaxation in rabbit aorta

Nicotinic receptor signaling in nonexcitable cells

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