7 Prostaglandins and Leukotrienes

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From: Endocrinology: Basic and Clinical Principles, Second Edition (S. Melmed and P. M. Conn, eds.) © Humana Press Inc., Totowa, NJ

7 Prostaglandins and Leukotrienes

Locally Acting Agents

John A. McCracken, PhD

CONTENTS

INTRODUCTION

H^ISTORICAL B^ACKGROUND PG NOMENCLATURE

BIOSYNTHESIS

EICOSANOID RECEPTORS

SELECTED EXAMPLES OF LOCAL ACTIONS OF EICOSANOIDS

A MODEL FOR ENDOCRINE CONTROL OF PULSATILE PGF_2α SECRETION DURING LUTEOLYSIS IN SHEEP

CONCLUSION

matory drugs underlines their importance in this regard.

This chapter includes a brief historical background, together with a description of nomenclature, biosynthesis, and selected local actions of PGs and LTs. For those requiring more detailed information, see the references at the end of this chapter.

2. HISTORICAL BACKGROUND

The biologic existence of PGs was established in the 1930s by the detection of smooth muscle contracting and vasodepressor activity in extracts of human and sheep seminal plasma. von Euler (1988) went on to demonstrate that these compounds were acidic lipids and the name prostaglandins was coined because it was thought, at the time, that these acidic lipids emanated from the prostate gland; however, we now know that the seminal vesicles are the main site of synthesis in the male reproductive system. It is not surprising that the biologic activity of PGs was first detected in the male system because they are present in microgram quantities in seminal plasma of many species including human, monkey, and sheep. By contrast, PGs in other tissues are present in picogram or, at best, nanogram

1. INTRODUCTION

This chapter is not intended for the prostaglandin (PG) specialist but, rather, for those not familiar with the PG field. It provides a brief overview of the biology of the eicosanoid family and how these local mediators may function in health and disease. The term eicosanoids (from the Greek eicosa, which means 20) was coined to describe the broad group of compounds derived from C₂₀fatty acids that, in turn, are derived from the essential dietary fatty acids. The predominant C₂₀ fatty acid precursor for eicosanoid biosynthesis in most mammals is arachidonic acid (AA). These eicosanoids include the PGs and thromboxanes (TXs), leukotrienes (LTs), lipoxins (LPXs), and hydroxyeicosatetraenoic acids (HETEs). Because the biologic activity of eicosanoids is diminished rapidly in both tissues and the circulation, it is likely that they act locally at the tissue and organ level, where they may regulate regional blood flow and other metabolic activities. Moreover, their formation in various inflammatory sites indicates an important mediating role for these substances in diseased states. Indeed, eicosanoid inhibition by different classes of anti-inflam-

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94 Part II / Hormone Secretion and Action

amounts. The function of PGs found in seminal plasma has not been fully documented, although a role has been suggested in contraction of the male accessory glands and the vas deferens. In addition, it has been proposed that they assist in sperm transport by stimulating contraction of the female genital tract, the latter also being a major source of PG synthesis and action. Indeed, the most fully established physiologic role of the PGs is that of PGF_2α, a locally acting uterine luteolytic hormone in a number of mammalian species (see Section 6.7.).

Progress in identifying the chemical nature of the PGs was slow, partly because of World War II and partly because the technology to detect these labile and elusive compounds was not available until the 1950s and 1960s. Bergström and Sjovall (1957) isolated two different PGs in crystalline form, one of which they named PGE, found in the ether (E) fraction, and the other PGF, found in the fraction with phosphate, spelled fosfat (F) in Swedish, thus giving rise to the present nomenclature. Samuelsson and colleagues (1987) discovered a related product of AA metabolism that is a potent stimulator of platelet aggregation and named it TX. Later, Vane and colleagues discovered a potent inhibitor of platelet aggregation derived from AA and formed in endothelial cells and named it prostacyclin (PGI₂) because of its double-ring structure.

Shortly thereafter, Samuelsson described a new class of AA metabolites from leukocytes, some of which have chemotactic properties and others that increase vascular permeability. These substances were named LTs. They are produced from AA by the action of the enzyme 5- lipoxygenase (5-LO). In some of the LTs, the amino acid cysteine is incorporated into the molecule to give rise to the cysteinyl LTs (see below). The LTs are con- sidered to be involved in inflammatory processes in which they most likely act synergistically with other mediators such as histamine, bradykinin, and PGs to produce the classic signs of inflammation described by Celsus: redness (rubor), heat (calor), swelling (tumor), and pain (dolor). The involvement of PGs in the inflammatory process is underlined by the effects of non ste- roidal anti-inflammatory drugs (NSAIDs) such as aspirin and indomethacin, which are potent inhibitors of PG biosynthesis via inhibition of cyclooxygenase (COX) activity. The NSAIDs, however, have little or no effect on the lipoxygenase pathway responsible for LT biosynthesis. Indeed, because NSAIDs so efficiently block PG synthesis, these drugs most likely amplify LT synthesis by diverting AA into the lipoxygenase pathway. Thus, in patients with asthma, in whom LTs have been identified as a major mediator of bron- choconstriction, the use of NSAIDs is contraindicated.

On the other hand, corticosteroids have a potent inhibi-

tory effect on phospholipase A₂(PLA₂) activity, thus markedly reducing the availability of AA as a substrate for both the COX and lipoxygenase pathways. As described later, corticosteroids also appear to inhibit the synthesis of the inducible form of the COX enzyme (COX-2). Corticosteroids are thus the most useful therapeutic agents for patients with asthma at present; however, drugs based on LT receptor antagonists and LT biosynthesis inhibitors have now been developed and are providing important alternatives and/or additions to long-term corticosteroid therapy (see Section 4.3.).

3. PG NOMENCLATURE

The major PGs of the two series are summarized as follows:

PGA₂ Dehydration product of PGE₂ PGB₂ Isomerization product of PGA₂

PGD2 Abundant in neural tissues, sleep inducing, allergic responses

PGE2 Vasodilator, gastric cytoprotection, pyrogenic, bone resorption

PGF_2α Vasoconstrictor, luteolysis and labor PGG₂ Endoperoxide intermediate

PGH₂ Endoperoxide precursor for PG synthesis PGI₂ Antiplatelet aggregation, vasodilator PGJ₂ Dehydration product of PGD₂ TXA₂ Platelet aggregation, vasoconstrictor

3.1. Chemical Structure

As illustrated in Fig. 1, the number of double bonds in the PG molecule is designated by a subscript so that the one series of PGs has one double bond in position 13:14, e.g., PGF_1α. The two series of PGs has a second double bond in position 5:6, e.g., PGF₂_α. The three series of PGs has a third double bond in position 17:18, e.g., PGF_3α. In the case of PGF, the α designation indicates that the hydroxyl groups are in the α orientation.

The principal PG precursor in most species is AA, lib- erated from phospholipid stores, principally by cytosolic PLA₂, which gives rise to the two series of PGs. The one series of PGs is derived from homo-γ-linolenic acid, which, in most species, is less abundant. Some species, especially when on diets rich in fish oils, produce the three series of PGs derived from eicosapentaenoic acid.

It is suggested that the high consumption of fish oils by Eskimos has a protective effect on the cardiovascular system in the presence of a high-fat diet.

4. BIOSYNTHESIS 4.1. Prostaglandins

A simplified flow sheet of the major pathways of eicosanoid biosynthesis is shown in Fig. 2. Virtually all cells appear to have the capacity to synthesize PGs, the

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end product depending on the enzymes present that convert the endoperoxide intermediates into specific PGs. The initial step in PG biosynthesis is the formation of AA from phospholipid stores via the action of cytosolic PLA₂. The microsomal COX which converts AA to the endoperoxide intermediates PGG₂ and PGH₂, is now known to exist in both a constitutive form (COX-1) and an inducible form (e.g., activated by serum or endotoxin; designated COX-2). The crystal structures of COX-1 and COX-2 are almost identical except for one amino acid difference that leads to a larger side pocket in COX-2 for substrate recognition.

Chandrasekaharan et al. (2002) have recently identified a variant derived from the COX-1 gene, designated COX-3, which is particularly sensitive to acetami- nophen and other antipyretic drugs (see Section 6.4.).

The NSAIDs such as aspirin or indomethacin act mainly by inhibiting the activity of the COX enzymes (also known as PGH endoperoxide synthases). Indeed, different NSAIDs have selective effects on the inducible form (COX-2) vs the constitutive form (COX-1). Cor- ticosteroids, which act primarily by blocking the phos-

pholipase-mediated release of AA from phospholipids, also have an additional inhibitory effect by blocking the formation of the inducible form of COX (COX-2), thus contributing to the blockade of PG synthesis.

Although it is well documented that PGs are an important component in acute inflammatory responses, it was observed that PGs of the E series may have certain modu- lating effects in some types of chronic inflammation;

that is, high tissue levels of PGEs may have anti-inflammatory effects. Weissmann (1993) and, more recently, Zurier (2003), who have studied the role of PGs in inflammation for many years, suggest that the proposed anti-inflammatory action of PGE₂ in certain chronic inflammatory states may be mediated by its ability to generate cyclic adenosine monophosphate (cAMP).

Because certain NSAIDs, such as sodium salicylate, can alleviate inflammation without inhibiting PG synthesis, it has been proposed that PGs may be modulators, rather than mediators, of inflammation (Weissmann, 1993).

Moreover, PGE₁ has been shown to suppress adjuvant disease (induced polyarthritis) in animal models and to inhibit neutrophil-mediated tissue injury (Zurier, 2003).

Fig. 1. Homo-γ-linolenic and arachidonic acids are converted, respectively, into prostanoids (PG) of the one series (PG1), exhibiting only one double bond, and into the two series (PG₂) exhibiting two double bonds. These polyunsaturated acids and their precursor, linoleic acid, are members of the biologic family of ω-6 fatty acids, characterized by an end segment of 6 carbons (at the opposite end from the —COOH). Eicosapentaenoic acid, coming from the α-linolenic acid (ω-3 family), is converted into PG3(three double bonds). Thick lines represent characteristic end segments of the ω-6 and ω-3 families. (Reproduced from Deby, 1988.)

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Since the 5-LO pathway is largely unaffected by NSAIDs, LT production may be potentially enhanced by diverting AA into the lipoxygenase pathway. In some instances, generation of LTs (e.g., the production of LTB₄by neutrophils), can be inhibited by PGs, suggesting that there may be a subtle balance between COX and lipoxygenase pathways. Moreover, both PGE₂and PGI₂ have been shown to downregulate the production of tumor necrosis factor-α (TNF-α).

It has been proposed that the administration of a com- bination of NSAIDs and long-acting PGE analogs could act synergistically in anti-inflammatory therapy (Weiss-

mann, 1993). It is likely that some of these opposing actions of PGE are related to the various PGE receptor subtypes that have now been elucidated and that can give rise to different second-messenger systems (see Section 5.1.).

4.2. Platelet-Activating Factor (Alkyl-acetyl-glycerophosphocholine)

Platelet-activating factor (PAF) is derived from phosphatidylcholine as a product of PLA₂action, although structurally it is not an eicosanoid. However, PAF is a potent platelet-aggregating substance that can operate Fig. 2. Simplified pathway of eicosanoid biosynthesis. PGFM = PGF_2α metabolite.

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without adenosine 5´-diphosphate or TXA₂. O’Flaherty and Wykle (2004) reviewed the biosynthesis of PAF in various tissues and concluded that PAF may act as both an intracellular and an extracellular mediator. As well as acting as an intracellular mediator, PAF appears to act in an autocrine fashion, i.e., by binding to the parent cell receptors, thus initiating other second-messenger signals. PAF appears to act in conjunction with endogenous eicosanoids with which it is often coformed during inflammatory states and allergic processes such as asthma. In addition, it has been suggested that PAF may play a local role during implantation of the embryo in the uterus, a process that also may involve PGs and other local mediators. A simplified chart showing the biosynthesis of PAF is shown in Fig. 3. In addition to the main PLA₂pathway, the PLC pathway is illustrated to show that activation of this signal transduction mechanism may also generate AA for eicosanoid formation.

Recent studies by Cundell et al. (1995) indicate that virulent pneumococci utilize the PAF receptor to gain entry into host cells. This bacterium is a commensal in the human nasopharynx and is a major cause of sepsis, pneumonia, and meningitis. Bacterial entry into endo-

thelial cells is increased 20- to 40-fold following stimu- lation of PAF receptors by fibrin or TNF-α and reversed by PAF receptor antagonists. It is suggested that bacterial cell wall phosphatidylcholine is a cognate ligand for the PAF receptor and that bacterial attachment subverts the receptor (in the absence of signal transduction) to internalize the bacterium. These novel findings suggest that PAF receptor antagonists may be of therapeutic value, not only in blocking PAF action, but also by attenuating bacterial attachment, which leads to inva- sion of host cells.

4.3. Leukotrienes

Like the PGs, LTs are considered to be local mediators generated in the microenvironment and usually associated with inflammation. The LTs are generated from AA released from membrane phospholipids or from secretory granules of tissue cells such as neutrophils or mast cells. The main enzyme in LT synthesis, 5-LO, requires activation by Ca⁺⁺and translocation to a membrane-associated site. These requirements are in contrast to PG synthesis in which COX does not require specific activation but, rather, only the presence of sub- Fig. 3. Simplified diagram of pathways for biosynthesis of PAF and eicosanoid products of AA metabolism. Note that PLA₂action forms both AA and PAF whereas the action of PLC can form lesser amounts of AA indirectly from diacylglycerol (DAG) via DAG lipase (e.g., during signal transduction). AA = arachidonic acid; FA = fatty acid.

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strate (AA). An additional regulatory component in LT biosynthesis is the existence of a 5-LO activating protein (FLAP) thought to be essential for LT biosynthesis.

Local synthesis of LTs is part of a cascade of events

occurring during inflammation that include the release of PGs, PAF, histamine, and other cellular mediators of this process. The LT pathway from AA involves the synthesis of LTA₄via the 5-LO pathway (Fig. 4).

Fig. 4. Metabolism of arachidonic acid by lipoxygenase pathways. (Reproduced from Piper and Letts, 1988.)

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LTA₄is nonenzymatically converted into LTB₄or into the cysteinyl LTs LTC₄, LTD₄, and LTE₄. Human neutrophils have the selective capacity to synthesize LTB₄ and also appear to inactivate LTB₄, thus regulating its local activity, which includes chemotaxis and neutrophil adherence to endothelial cells. Eosinophils, on the other hand, generate only LTC₄because they possess LTC₄synthase, whereas monocytes generate both LTB₄ and LTC₄. Cells lacking the enzymes required to produce specific eicosanoids may utilize an intermediate provided by another cell type. For example, the transfer of PG endoperoxide intermediates from platelets to endothelial cells results in the formation of PGI₂, which has antiplatelet aggregating activity. Erythrocytes lack 5-LO but can convert LTA₄into LTB₄. These cell–cell interactions illustrate the complexity of eicosanoid synthesis likely to occur, e.g., in inflammatory processes.

The three cysteinyl LTs (LTC₄, LTD₄, and LTE₄) are now known to constitute the slow-reacting substance of anaphylaxis, and they are implicated in the pathogenesis of asthma and other pulmonary conditions. Inhalation of LTD₄and LTC₄by humans is 1000 times more potent than histamine in causing airflow impairment at a fixed vital capacity, whereas LTE₄is 10 times as potent as histamine. The involvement of LTs in anaphylaxis and asthma has prompted the synthesis of a number of LT receptor antagonists including Singulair (montelukast) and Accolate (zafirlukast), which are presently in clinical use. Another approach is the development of the LT biosynthesis inhibitor Zyflo (zileuton) which has no direct effect on the 5-LO enzyme itself but binds with high affinity to FLAP, whose expression is required for LT biosynthesis. The development of these new LT-inhibiting drugs, including those binding to FLAP, will most likely help to alleviate the clinical symptoms associated with asthma and other pulmonary conditions, thus providing an alternative, or an addition, to corticosteroid therapy (Robin- son et al., 2001).

Arachidonic acid can also be converted via other lipoxygenase pathways to yield a series of hydroxyeicosatetraenoic acid derivatives (HETEs) including lipoxins (from “lipoxygenase interaction substances”) and other HETEs (Fig. 2). These lipoxygenase products are considered to be associated with the immune system, as proposed by Vanderhoek (1992), where they may play a role as endogenous immunosuppressive agents. Recent studies by Klein and colleagues (2004) have indicated a potential role for 12/15 lipoxygenases in the pathogenesis of osteoporosis in a mouse model.

Their findings suggest that inhibitors of the 12/15 lipoxygenase pathway, already in clinical use, may merit investigation for the prevention and/or treatment

of osteoporosis. Arachidonic acid also can be metabo- lized to epoxides by cytochrome P-450 (Capdevila and Falck, 2001). The functional role of epoxides is only now being explored but may include control of systemic blood pressure and the pathophysiology of hypertension.

Because eicosanoids are released from tissues as soon as they are formed, they are not stored within the cell.

However, in the case of the male reproductive system, PGs accumulate in large amounts in the glandular secretions found in the lumen of the seminal vesicles. Most tissues metabolize PGs very rapidly via the 15-hydroxy dehydrogenase/13:14 reductase complex, which is particularly abundant in lung tissue, thus forming the biologically inactive metabolite (e.g., PGFM from PGF_2α).

In many species, one passage through the lungs can inactivate >90% of the biologic activity of the primary PGs. The net effect is that, whereas returning venous blood can contain considerable amounts of PGs in the nanogram range, aortic blood contains much lower levels, probably in the low picogram range. Thus, PGs are generally regarded as locally acting agents because their very potent biologic activity is necessarily restricted by 15-hydroxy dehydrogenases, both at the tissue level and in the vascular system, by pulmonary metabolism and, to some extent, by metabolism in the liver and kidney.

4.4. Isoprostanes

Roberts and Morrow (1994) have identified a novel series of PG-like compounds, called F₂-isoprostanes, that are formed as products of lipid peroxidation of membrane phospholipids catalyzed by free radicals, (i.e., independent of the COX pathway). One of the isoprostanes, 8-epi-F_2α, has been identified as the most potent renal vasoconstricting substance ever discovered.

The marked vasoconstrictor effect of this compound has been shown to be mediated by activation of TX receptors. Current work suggests that the overproduction of isoprostanes may play a causative role in the hepatorenal syndrome, defined as unexplained renal failure in the presence of severe liver disease. Thus, in addition to being markers of lipid peroxidation, it is likely that isoprostanes may be associated with the pathophysiology of oxidant stress, suggesting that antioxidant therapy may provide a new rationale for therapeutic intervention in certain disease states.

5. EICOSANOID RECEPTORS

PGs, TXs, and LTs are considered to act locally via specific receptors located on the cell surface. However, the specificity of these receptors shows considerable overlap, so that responses to PGs within different tissues may vary or even have opposite actions. The nature

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100 Part II / Hormone Secretion and Action

and affinities of receptors for various PGs were initially studied using radiolabeled ligands. PG receptors have also been characterized pharmacologically by compar- ing the potencies and responses of various PGs and their analogs in a variety of bioassay systems. Based on these findings, Coleman and colleagues (1994) have described a pharmacologic classification in which each PG has its own receptor, some of which show subtypes. In the case of PGE, four subtypes have been assigned and designated EP1, EP2, EP3, and EP4. Evidence has also accumulated that TXA₂ has two receptor subtypes, designated TPα and TPβ, which regulate platelet aggregation and vascular smooth muscle contraction, respectively.

Narumiya and Fitzgerald (2001) have pioneered the cloning of the PG receptors, the first of which was TXA₂. This was accomplished by isolating and purify- ing it from human blood platelets followed by cloning of the cDNA. These studies indicated that the TXA₂ receptor is a G protein–coupled rhodopsin-type receptor with seven transmembrane domains. Because there is only a 10–20% homology with other rhodopsin-type receptors, it is proposed that TXA₂ and other prostanoids constitute a subfamily of the rhodopsin- type receptor superfamily. Based on this model, screen- ing of mouse cDNA libraries revealed seven different

prostanoid receptors. Four receptor subtypes of the PGE receptor were cloned and designated EP1, EP2, EP3, and EP4, thus confirming the aforementioned pharmacologic classification. In addition to the receptors encoded by different genes, alternative splicing of the EP₃receptor transcript has yielded three isoforms of the receptor that couple to various G proteins and induce specific signaling systems (Fig. 5). The PGD receptor exists in two forms, DP1 and DP2, with each receptor type derived from a single gene. DP1 is thought to be involved in mediating allergic asthma and DP2 is assigned a role in lymphocyte chemotaxis.

Recently, two forms of the FP receptor have been cloned, designated FP_A and FP_B (Sakamoto et al., 2002). Although the role of the conventional FP receptor, FP_A, is well defined, the role of the new FP receptor, FP_B, remains to be investigated. The so-called relaxant receptors—IP, DP, EP2, and EP4—are considered to act via G protein–mediated increases in cAMP, and the so-called contractile receptors—EP1, FP, and TP—are considered to act via a G protein–

mediated increase in intracellular Ca⁺⁺. By contrast, the EP3 receptor is thought to be the exception, because it decreases cAMP via G-protein interaction.

Although many PG receptors are located in the plasma membrane, there is also evidence that some PG Fig. 5. Seven-transmembrane-spanning model of mouse EP3 isoforms showing alternative splicing for isoforms. CHO indicates potential sites of N-linked glycosylation. Stars indicate potential phosphorylation sites for cAMP-dependent protein kinase. (Repro- duced from Negishi et al., 1995.)

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receptors are located in the nuclear envelope. The finding that COX-2 is predominantly located near the peri- nuclear envelope thus provides a convenient interaction with the PGs so formed, with receptors in the nuclear envelope to effect increases in Ca⁺⁺and gene transcription. LTs also act via G protein–coupled receptors, several of which have been characterized.The LTB4 receptor (B-LT) exists in two forms, one regulating chemotaxis of leukocytes and the other regulating neutrophil secretion (Yokomizo et al., 2000).

Receptors also exist for the cysteinyl LTs, LTC4 and LTD4, designated cysLT-1 and cysLT-2, respectively, and are thought to mediate the action of these LTs in the pulmonary and vascular beds. PGs and LTs may also interact with peroxisome proliferator-activated receptors, members of the nuclear superfamily that includes steroid hormone receptors. PGI₂and PGD₂and its metabolite PGJ₂, but not PGE₂or PGF₂_α, have been shown to activate peroxisome proliferator-activated receptors.

5.1. PG Transporter

Some years ago, McCracken and colleagues showed that during luteolysis in sheep, PGF_2αdiffuses from the uterine vein rather slowly through the wall of the closely adherent ovarian artery. This countercurrent mechanism allows about 1% of PGF_2α secreted by the uterus to escape metabolism by the lungs and reach the ovary directly, where it causes luteolysis (see Section 6.7.). In some tissues, PG transport appears to be enhanced by a carrier-mediated transporter that has been identified in epithelial cells (see Banu et al., 2003) as a protein with 12 transmembrane domains. Because the PG transporter appears to facilitate transport of PGs across cell membranes, it does not appear to be a receptor as such, particularly because it differs from the seven-transmembrane structure of the PG receptors. The PG transporter is thought to mediate the uptake and release of newly synthesized PGs from cells and to facilitate vectoral transport. Recently, high concentrations of the PG transporter have been identified in the bovine uterine vein/

ovarian artery complex as well as other reproductive tissues (Banu et al., 2003). Such a finding may explain the mechanism underlying the countercurrent transfer of PGF_2αthat occurs in the sheep and cow and several other species. Some PGs, such as PGI₂and its metabolite, 6-keto-PGF_1α, show very poor interaction with the PG transporter, which may explain the much-reduced metabolism of these compounds in the pulmonary circulation. The biologic activity of other PGs present in venous blood is rapidly reduced by metabolism in one passage through the lungs, presumably because the PG transporter very efficiently allows access to the intrac-

ellular 15-hydroxydehydrogenases present in the lung parenchyma. This explanation is consistent with the finding that, although PGI₂is indeed a substrate for 15- hydroxydehydrogenase in cell-free systems, it probably escapes metabolism in the lungs by virtue of its very weak interaction with the PG transporter. However, it should be emphasized that PGI₂in vivo is intrinsically unstable, being transformed nonenzymatically to its inactive metabolite, 6-keto-PGF_1α, in about 20 s. Thus, it is a “local hormone” rather than a circulating one because arterial levels (about 3 pg/mL) are probably too low to have any systemic effect. LTC₄is also known to be transported extracellularly by transporters such as multidrug resistance–associated protein, and, in this way, LTC₄is thought to mediate the migration of den- dritic cells to lymph nodes (Robbiani et al., 2000).

6. SELECTED EXAMPLES OF LOCAL ACTIONS OF EICOSANOIDS

6.1. Gastric Cytoprotection

PGE₂appears to be an endogenous inhibitor of gastric acid secretion and to have a cytoprotective effect on the gastric mucosa. The ingestion of aspirin, indomethacin, and other NSAIDs that inhibit PG synthesis can cause severe damage to the gastric mucosa, with formation of ulcers and gastric bleeding. That inhibition of PG synthesis leads to damage to the mucosa is demonstrated by the coadministration of NSAIDs and PGE₂, or one of its various analogs, which prevents mucosal damage. Flower (2003) has recently evaluated the ability of several different NSAIDs to inhibit the two forms of COX, COX-1 (constitutive) and COX-2 (inducible) and has reviewed in detail the evolution and development of selective inhibitors for COX-2. The amino acid sequence of COX–2 shows a 60% homology with COX-1, but both enzymes have the same mol wt, about 70 kDa, and possess similar active sites for binding NSAIDs. However, the side pocket differs by one amino acid and results in a larger target for COX-2 inhibitors.

Aspirin and indomethacin show greater inhibition of COX-1 (the predominant form in the gastric mucosa) than COX-2, which is consistent with the relative pro- pensity among various NSAIDs to cause gastric ulcer- ation. Thus, it has been possible to design NSAIDs (selective COX-2 inhibitors) that have minimal effects on gastric COX-1, but that inhibit the COX-2 induced by inflammatory mediators in other tissues. Such drugs, which include Celebrex and Vioxx (withdrawn from market 2004), permit safer oral administration for maximal systemic effects with diminished gastric side effects, which are reduced by at least 50%. However, since some of these drugs may inhibit COX-2 in

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endothelial cells, they could potentially skew the ratio of PGI₂to TXA₂and influence blood platelet aggregation. An unusual example of local action of PGs on gastric function is provided by a species of Australian frog that incubates its eggs in the stomach. Extremely high levels of gastric PGE₂during the incubation are thought to act locally to inhibit gastric acid secretion and to promote gastric mucous secretion, thus protecting the eggs against digestion in the stomach.

The local production of LTs may also play a role in gastric function because ethanol-induced mucosal damage in the rat is accompanied by an increase in LTC₄. Moreover, nordihydroguaiaretic acid, a nonspecific lipoxygenase inhibitor, prevents ethanol-induced gastric mucosal damage and, at the same time, inhibits mucosal release of LTC₄. Thus, endogenous local production of PGE₂has a protective effect on the stomach, although the effect is blocked by NSAIDs, whereas LTs appear to be local mediators of ethanol-induced damage to the gastric mucosa.

6.2. Local Action of PGs in Glaucoma

A biologically active substance was isolated from the iris and was found to increase during ocular inflammation. This substance, which caused a marked constriction of the pupil, was initially named irin. Subsequently, the biologic activity of irin was shown to be owing to PGs, principally PGF_2α. This is consistent with previous findings that NSAIDs were effective in reducing inflammation in the eye. Paradoxically, it was found later that PGF_2αapplied topically to the eye reduced intraocular pressure, suggesting a physiologic role for PGs in regulating intraocular pressure. Stjernschantz and colleagues (2000) pioneered the development of PGF_2αanalogs for the treatment of glaucoma, which is characterized by a chronic increase in intraocular pressure. PGF_2αwas found to reduce intraocular pressure, not by increasing outflow via the trabecular meshwork or by reducing the production of aqueous humor but, rather, by increasing the uveoscleral outflow of aqueous humor through the ciliary muscle. Some of these analogs, several of which are now in clinical use, have additional effects on the microvascu- lature of the eye and may increase capillary permeability.

Although some modest side effects of daily topical appli- cation of PGF_2α analogs are observed, the dramatic and long-lasting reduction in intraocular pressure indicates that further development of PGF_2αanalogs may provide more clinically acceptable treatments for glaucoma.

6.3. Local Effect of PGD

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in Sleep Induction

For many years, Hayaishi and colleagues (1993) studied the role of PGD₂production in the preoptic area (POA)

of the brain in relation to the induction of physiologic sleep. They found that microinjection or infusion of PGD₂ in femtomolar concentrations into the POA (an area considered to be the sleep center), or into the third ventricle of the rat or monkey, was effective in inducing normal sleep. They went on to show that the specific activity of the enzyme that controls PGD₂production in the brain, PGD₂synthase, exhibits a circadian fluctuation that par- allels the sleep/wake cycle. When PGD₂synthase activity in the POA was inhibited by the infusion of a specific inhibitor, selenium, sleep was inhibited. Such inhibition was reversed when the infusion of selenium into the POA was stopped. Hayaishi and colleagues (1993) conclude that PGD₂is produced and acts locally in the POA of the brain as the physiologic inducer of sleep.

6.4. Mediating Role of PGE

₂

in Fever

Evidence has accumulated over several years that PGE₂is a primary mediator of fever induced by bacterial pyrogens. Skarnes and colleagues (1981) investigated the role of PGE₂in endotoxin-induced fever in conscious sheep. They showed that PGE₂infused into the carotid artery caused a transient increase in blood pressure (10–

20%) accompanied by a sustained (3 h) increase in core body temperature that mimicked the effects seen during the first wave of fever induced by intravascular endo- toxin (see Fig. 6). Subsequent experiments revealed that during the first phase of endotoxin-induced fever, a marked increase in both PGE₂and PGF_2αoccurred in the venous and arterial circulation. Intracarotid PGF₂_αitself, however, did not exhibit the pyrogenic or pressor effects seen with intracarotid PGE₂. Further work showed that indomethacin blocked both the first and second phases of fever induced by endotoxin. More important, PGE₂ evoked both the pressor response and the first phase of fever during indomethacin blockade. It was concluded that PGE₂, formed within the vasculature, crosses the blood-brain barrier and acts on the thermoregulatory center in the hypothalamus to evoke the first phase of fever.

However, since both the first and second phases of fever are blocked by indomethacin, it appeared that the presence of circulating levels of another AA metabolite might be responsible for the second phase of fever (i.e., the phase associated with leukocyte pyrogen, interleukin-1 [IL-1]). Studies by Coceani and colleagues (1989), using the cat as a model, supported the role of PGE₂in the pathogenesis of fever induced by bacterial pyrogen. By means of a push-pull perfusion procedure, they demonstrated that iv endotoxin selectively caused an increase in PGE₂in the anterior hypothalamic/preoptic region of the brain. The increase in PGE₂production in these areas of the brain was consistent with the onset and progression of fever in their experimental model.

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Recent work using knockout mice lacking different subtypes of the EP receptor supports and extends the key role of PGE₂in endotoxin-induced fever. Sugimoto and colleagues (2000) showed that knockout mice defi- cient in the receptor subtypes EP1 and EP2 continued to exhibit a fever response after an endotoxin challenge but that no fever occurred with endotoxin in EP3 knockout mice. This same group went on to demonstrate that local synthesis of PGE₂, induced by lipopolysaccharide or a cytokine, interacts with EP3 receptors located in the neurons surrounding the organum vasculosa lamina terminalis (OVLT), considered to be the thermoregulatory center, thus inducing a fever response to endotoxin.

It now seems likely that, as demonstrated in the sheep model, the initial wave of fever and pressor responses induced by endotoxin is mediated by the rapid increase in the general circulation of PGE₂, which readily passes across the blood-brain interface to act on the themo- regulatory center in the hypothalamus as well as on the vasomotor center in the medulla. The fever response to PGE₂ is very specific because, in the sheep model, intracarotid PGD₂causes a pressor response but does not induce fever. The second and more sustained wave of fever observed in the sheep and other species may be owing to the induction of COX-2 and/or COX-3 in the OVLT by cytokines such as IL-1.

Fig. 6. Effect of indomethacin treatment (4.0 mg/kg, subcutaneously) on fever and pressor responses to an intracarotid infusion of endotoxin (ET) (10 μg/min) or PGE2(2.6μg/min). Indomethacin blocked both the first and second waves of fever and the pressor response to endotoxin, but the first wave of fever and the pressor response to PGE₂were elicited during indomethacin treatment. BL. PRES. = blood pressure. (Adapted from Skarnes et al., 1981.)

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Moreover, recent studies by Engblom and colleagues (2003) provide further support for a role of centrally generated PGE₂in the pathogenesis of fever.

Using knockout mice lacking the gene for microsomal PGE synthase-1, they demonstrated that these mice did not show a febrile response to endotoxin injected intraperitoneally, whereas pyresis occurred in these same animals in response to PGE₂injected into the cerebral ventricles. Thus, the delayed increase in the local generation of PGE₂in the OVLT would provide an answer for the second wave of fever. Such a scenario would explain why indomethacin blocks the first wave of fever caused by peripherally generated PGE₂as well as blocking the second wave of fever mediated by centrally generated PGE₂. The initial pressor response to intracarotid PGE₂, as well as to intracarotid endotoxin infusion, is consistent with the view that an increase in PGE₂in the general circulation, caused either directly by PGE₂infusion or indirectly by endotoxin, causes a central, peripherally mediated increase in arterial pressure. This most likely explains why a pressor response occurs with endotoxin only during the first wave of fever. The second wave of fever induced by endotoxin, associated with a delayed local increase in PGE₂ in the OVLT, thus does not result in a pressor response. The identification of a third form of COX, COX-3 (Chandrasekharan et al., 2002), particularly in neural tissues, together with the high sensitivity of COX-3 to inhibition by acetami- nophen, supports a central role for neural COX-3 in the generation of the second sustained wave of fever.

6.5. Pheromonal and Reproductive Function of PGs in Teleost Fish

Among the more unusual actions of PGs is their recently proposed role as pheromones in male teleost fish. Sorensen and Goetz (1993) have investigated this phenomenon. PGF_2αand/or its metabolites are produced in large quantities in the ovaries of female teleost fish, where they appear to act in a paracrine fashion by stimulating follicular rupture, a phenomenon also observed during ovulation in most mammalian and nonmamma- lian species. In female teleost fish, PGF₂_α and/or its metabolites secreted by the ovary persist in the systemic circulation (since fish do not have lungs) and act centrally in the brain to elicit female spawning behavior.

Apparently the female fish releases into the water during oviposition relatively large amounts of PGF metabolites that are detectable by the highly developed chemosensory system in the male fish. The male is thus “signaled” through a sensitive olfactory system to exhibit spawning behavior.

6.6. Local Action of PGs in Uterus

Although PGs were first isolated and identified from secretions of the male reproductive tract, it was not long before they were also identified in the female reproductive system, where they serve a number of important local functions. Key findings included the discovery that human menstrual fluid was a major source of PGs, particularly PGE₂and PGF_2α, and that the administration of PGF₂_α to women during the luteal phase produced menstrual-like bleeding. Moreover, Eldering and colleagues (1993) reported high production rates of several PGs (especially PGF_2α) from endometrium of the rhesus monkey, primarily during the luteal phase.

These findings led to extensive studies on the role of PGs in normal menstruation and in abnormal conditions such as dysmenorrhea and menorrhagia.

Bygdeman and Lundstrom (1988) investigated the role of PGs in menstrual physiology. It is established that an overproduction of endometrial PGs explains the symptoms of dysmenorrhea in women. This conclusion is supported by the finding that a variety of NSAIDs alleviate the clinical symptoms of dysmenorrhea (uterine cramping) and menorrhagia (excessive blood loss). The effectiveness of different NSAIDs varies considerably, owing to the fact that in addition to blocking COX activity in the uterus and hence PG production, some of these compounds (such as the fenamates) also act as PG receptor antagonists. Intrauterine devices (IUDs) stimulate the local production of endometrial PGs, which may mediate the mechanism of action of IUDs in preventing conception. In some individuals, the presence of an IUD results in an overproduction of PGs and the occurrence of dysfunctional bleeding, which often can be controlled by NSAIDs.

6.7. Identification of PGF

_2α

as a Local Uterine Luteolytic Hormone

The occurrence of PGs in human menstrual fluid and their role in menstruation provided an important lead regarding the potential role of PGs in uterine physiology in mammals. Unlike in primates, the presence of the uterus is essential for cyclic regression of the corpus luteum in many nonprimate species. McCracken and colleagues (1972) investigated the role of uterine PGs in relation to cyclic regression of the corpus luteum using sheep with autotransplanted ovaries and/or uterus.

The presence of a contiguous uterine horn was found to be a requirement for luteolysis in this species. Such a requirement was subsequently explained by the local uterine production and countercurrent transfer of PGF_2α from the uterine vein to the ovarian artery, the latter vessel being highly contorted and adherent to the uter-

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ine vein (see Fig. 7). This mechanism allows PGF_2α secreted by the uterus to avoid metabolism in the pulmonary circulation and to reach the adjacent ovary directly, hence causing luteolysis. The PG transporter, recently identified in bovine reproductive blood ves- sels, most likely accounts for the countercurrent transfer process. Such a bypass mechanism from vein to artery allows the transfer of about 1% of the secreted amount of uterine PGF_2α(see Fig. 8), which occurs as a series of episodic bursts during luteolysis. It is likely that similar countercurrent transfer mechanisms for PGs and other substances may occur where the appropriate vascular anatomy exists, such as in the testicular vascular pedicle or in the kidney vasculature. Indeed, the transfer of tritiated water, krypton, and heat is reported to occur in the testicular vasculature. In addition, ste-

roids are known to be transferred within the ovarian vasculature of several species including the human.

Such a transfer mechanism may permit steroids and other substances to reach the ovary, the oviduct, and perhaps the uterus itself in a concentration greater than would be supplied via the systemic circulation.

6.8. PGs and Cancer

Evidence has accumulated in recent years that the use of NSAIDs decreases the incidence of certain cancers, especially colorectal, breast, and lung cancer. Moreover, NSAIDs suppress carcinogen-induced cancers in several animal models. A number of potential targets for NSAID action in the prevention of cancer have been proposed. In a recent study, Lui and colleagues (2001) showed that overexpression on COX-2 in transgenic Fig. 7. Anatomic relationship of ovarian artery and uteroovarian vein in sheep and experimental design used to demonstrate coun- tercurrent transfer of ³HPGF_2αfrom a uteroovarian vein to its closely adherent ovarian artery. About 1% of PGF_2αsecreted by the uterus reaches the ovary directly, thus bypassing metabolism in the pulmonary circulation. (Reproduced from McCracken et al., 1972.)

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mice is sufficient to induce tumorigenesis, suggesting that overproduction of PGs may promote cancer in certain tissues. Indeed, mammary tumors from these transgenic mice contained elevated levels of PGF_2α, PGE₂, PGD₂, and 6-keto-PGF₁_α. Moreover, studies have indicated an involvement of the EP1 receptor subtype in precancerous lesions evoked by chemical carcinogens (Watanabe et al., 1999).

The extracellular matrix (ECM) has also been implicated in the progression of tumorigenesis, potentially via changes in gene expression, basement membrane formation, cell adhesion characteristics, and cell migration (Liotta and Kohn, 2001). Recently, PGF_2α was shown in vivo to reduce acutely protein levels of tissue inhibitors of metalloproteinases (TIMPs), primary regulators of the ECM, in luteal tissue by as much as 90%

within 1 h of PGF_2αadministration (see Fig. 9) and then to increase metalloproteinase-2 activity (Towle et al., 2002). Such a rapid reduction in TIMPs in luteal tissue may be the initial step in functional luteolysis, e.g., by altering the conformation/localization of luteinizing hormone (LH) receptors on luteal cells. The rapid effect of physiologic levels of PGF_2αon regulators of the ECM in luteal tissue in vivo suggests one possible explanation for the tumorigenic effect of PGs since the unexplained beneficial effects of NSAIDS on certain cancers may be attributed to their attenuation of PG action on the ECM.

It may seem paradoxical that PGs (specifically PGF_2α)

promote regression of luteal tissue, whereas there is evidence that PGs promote tumorigenesis in susceptible tissues and that NSAIDs prevent it. However, recent work has shown that, in some tissues, TIMP-2 sup- presses TKR growth factor independently of metalloproteinase inhibition (Hoegy et al., 2001). Thus, acute suppression of TIMP-2 by a PG, as occurs in luteal tissue in vivo, may increase growth factor activation in certain tissues and, hence, contribute to tumorigenesis (McCracken, 2004).

6.9. NSAIDs and Alzheimer Disease

In recent years, several epidemiologic studies have indicated that the incidence of Alzheimer disease is reduced in individuals undergoing long-term NSAID therapy. Alzheimer disease is characterized by a buildup in the brain of amyloid plaques consisting of protein fibers and amyloid-β surrounded by astrocytes and microglia. The beneficial effects of NSAIDs on Alzheimer disease were considered initially to be owing to their antiinflammatory action via COX inhibition.

However, a specific explanation for the protective effect of NSAIDs against Alzheimer disease has been lacking. A recent study by Weggen and colleagues (2001) has indicated that the beneficial action of NSAIDs in Alzheimer disease may be owing to the reduction of amyloid-β by a mechanism independent of COX inhibition. Using mice engineered to provide a Fig. 8. Time course of countercurrent transfer of ³HPGF_2αfrom a uteroovarian vein to its closely adherent ovarian artery. (Repro- duced from McCracken et al., 1972.)

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model for Alzheimer disease, as well as cultured cells, these investigators showed that ibuprofen lowered the level of amyloid-β42, which has been implicated in the pathogenesis of this disease, by as much as 80%. It seems that the beneficial effect may be limited to certain NSAIDs, since aspirin therapy, as well as selective COX-2 inhibitors, has a much less convincing effect on the incidence of Alzheimer disease. Moreover, in Weggen et al.’s (2001) study, aspirin did not block the formation of amyloid-β42 in mice. Because the action of certain NSAIDs in lowering amyloidogenic proteins is independent of anti-COX activity, the development of similar compounds lacking COX inhibition could give rise to a new generation of drugs that selectively reduce brain amyloid-β42 formation without the side effects of long-term COX inhibition, such as gastric and renal side effects. On the other hand, the anti-inflammatory action of NSAIDs may help to reduce the amyloid plaque-induced inflammatory response, that occurs in Alzheimer disease.

7. A MODEL FOR ENDOCRINE CONTROL OF PULSATILE PGF

_2α

SECRETION

DURING LUTEOLYSIS IN SHEEP

Hormonal regulation of uterine PGF_2α secretion, which initiates luteolysis in nonprimate species, has been studied extensively in the sheep. This species offers a number of advantages as a model, such as the access to

the uterine and ovarian circulation in sheep bearing a transplanted ovary and/or uterus. Early studies indicated control of uterine PGF_2α secretion by estrogen and progesterone. However, McCracken and colleagues (1995; 1999) subsequently showed that these hormones act indirectly by regulating the formation of oxytocin receptors (OTRs) in the endometrium, the site of uterine PGF_2αsynthesis in sheep. A model for the cellular control of PGF₂_αsynthesis in the endometrium of the sheep is shown in Fig. 10. Estradiol-17β (E2) promotes the formation of endometrial OTRs in 6–9 h and may also increase PG synthetase and phospholipase levels. When OTRs are present on the endometrial cell membrane, oxytocin (OT) induces the immediate secretion of PGF_2α. Signal transduction via the phosphatidylinositol-mediated increase in intracellular Ca⁺⁺is thought to activate PLA₂, resulting in liberation of AA from phospholipid stores. Some AA may also be generated from DAG via DAG lipase. Released AA is then rapidly converted into PGF_2αby PG synthetase (COX). Progesterone secreted during the luteal phase initially inhibits estrogen action and prevents the formation of endometrial OTRs. Dur- ing this period of inhibition, progesterone appears to increase lipid stores in the endometrial cells (the so-called priming effect of progesterone). Finally, progesterone action diminishes toward the end of the luteal phase via catalysis of its own receptor. Such a loss of progesterone action will facilitate the return of E₂action with the con- Fig. 9. In vivo temporal changes in peripheral plasma progesterone and luteal protein levels of TIMP-1, TIMP-2, and MMP-2 following administration of a 1 h luteolytic pulse of PGF_2αto sheep during midcycle; n = three sheep per time point. (Adapted from Towle et al., 2002.)

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sequent formation of endometrial OTRs and, hence, the ability of OT to stimulate endometrial PGF_2αsecretion.

However, because of the effects of progesterone priming, the amount of PGF_2αsecreted in response to OT is 50- to 100-fold greater than the response to OT before priming.

The model proposed in Fig. 10 explains the cellular regulation of PGF₂_αsynthesis via the induction of endometrial OTRs, but it does not explain the role of circulating levels of OT. In the sheep and other ruminants, the regulation of blood levels of OT is complicated by the fact that, in addition to the neurohypophysis, the corpus luteum is an ectopic site of OT synthesis. Recent work from our laboratory has confirmed that a finite store of OT exists in the ovine corpus luteum that can be released by subluteolytic levels of PGF_2α. This supplemental source of OT appears to reinforce periodic signals from the central OT pulse generator to amplify the magnitude of each luteolytic pulse of PGF_2αfrom the uterus. A model of the endocrine control of pulsatile secretion of PGF_2αfrom the uterus during luteolysis in the sheep is depicted in Fig. 11 The numbers in the Fig.11 are explained as follows: (1) At the end of the cycle, loss of progesterone (P) action by catalysis of its receptor, in both the hypothalamus and the uterus, results in return-

ing E₂action. (2) E₂(E) increases endometrial OTRs and causes intermittent increases in the frequency of the central OT pulse generator. (3) Low-level PGF_2αis generated by the action of central OT on uterine OTR. (4) This low-level PGF_2αacts on the high-affinity PGF_2α receptor (HFPR) present on OT-containing luteal cells to stimulate OT secretion. (5) A high level of uterine PGF₂_αsecretion is now generated by luteal OT. (6) This high level of PGF_2αnow has two effects via the low- affinity PGF_2αreceptor (LFPR) in luteal cells: first, it stimulates additional luteal OT; and, second, it depresses luteal P secretion (luteolysis). The closed-loop positive feedback between the uterus (source of PGF_2α) and the corpus luteum (source of OT) will continue until PGF_2α receptors in the corpus luteum are desensitized, thus terminating the release of luteal OT. Additional luteolytic pulses will follow when luteal PGF₂_αrecep- tors and/or uterine OTR recover after 6–9 h, in time to respond to the next high-frequency signal from the central OT pulse generator. However, further work will be required to determine whether a luteal contribution of OT is an absolute requirement for the luteolytic process in sheep and other ruminants. A minimum of five luteolytic pulses of PGF_2αare necessary to complete functional luteolysis.

Fig. 10. Model for the endocrine control of PGF_2αsynthesis in endometrial cell of sheep during luteolysis (see text for explanation).

(Reproduced from McCracken et al., 1995.)

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In the past few years, receptors for LH have been identified in the endometrium and other nongonadal components of the reproductive tract of several species.

In the bovine species, LH receptors have been located in both the endometrium and the uterine veins, and LH stimulates the expression of COX-2 and the production of PGF_2αby these tissues (Shemesh et al., 1997). How- ever, a potential role for uterine LH receptors in the process of luteolysis remains to be investigated.

In the sheep and other nonprimate species, in addition to the well-established interplay between ovarian steroids and the hypothalamic/anterior pituitary axis for the initiation of the ovarian cycle (via the gonadotropins), there is now good evidence for an interplay between ovarian steroids and the hypothalamic/posterior pituitary axis for the termination of the reproductive cycle (via OT). In species that do not synthesize OT in the ovary, it seems likely that OT secreted via the central OT pulse generator may alone be a sufficient stimulus for the generation of pulsatile luteolytic releases of uterine PGF₂_αas in the mare and sow. Thus, in sheep and several other nonprimate species, the uterus can be regarded as a transducer that, under appropriate hormonal conditions, converts neural signals into local hormonal signals from the uterus to effect luteolysis. Although the uterus is an abundant source of PGs in primates, the presence of the uterus is not required for luteolysis because ovarian cyclicity is normal in hys- terectomized subjects. However, many of the components present in the sheep uteroovarian system are present within the primate ovary itself, namely small amounts of

OT, OTRs, PGF_2α, PGF_2αreceptors, and different luteal cell types. Thus, a local production and action of PGF₂_α, or another metabolite of AA within the primate ovary itself, could account for luteolysis, but, this possibility is still under investigation.

The endocrine control for the local production of PGF_2αin the endometrium at the time of luteolysis may also occur at the end of pregnancy during the induction of labor in a number of species. Work by Fuchs and colleagues (1992) indicates an upregulation of OTRs in the endometrium and in the placental membranes in the bovine fetoplacental unit prior to the onset of labor.

This upregulation of OTR is associated with the local formation of PGF_2αin these tissues. It is proposed that PGF_2α, so formed, acts synergistically with low levels of OT on the myometrium to promote uterine contrac- tions and the initiation of first-stage labor. The subsequent increase in maternal OT levels during the second or expulsive phase of labor is most likely owing to release of pituitary OT via the Ferguson Reflex (see Fuchs et al., 1992).

Another important local function of PGs during pregnancy is the maintenance of ductus arteriosus patency in the fetus. Momma (2004) investigated the role of PGs in ductal patency and concluded that PGE₂is the prime agent in this respect. PGE₂was found to be more than 1000 times more potent than PGI₂in maintaining ductal patency. The levels of PGE₂in the fetal circulation are 300–400 pg/mL of plasma vs 5–10pg/mL in the adult sheep. One likely explanation for the high levels of PGE₂ Fig. 11. Model for neuroendocrine control of pulsatile PGF_2αsecretion from uterus during luteolysis in sheep (see text for explanation of numbers in the model). (Reproduced from McCracken et al., 1995.)

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in the fetal circulation is that the low blood flow through fetal lungs may result in diminished pulmonary metabolism of PGE₂. Thus, it appears that ductal patency is maintained by high levels of PGE₂in the fetal circulation rather than by local mural production. Because NSAIDs and corticosteroids block PG synthesis, they can be used to effect ductal closure in the newborn and in cases of fetal prematurity. These drugs are contraindicated in women during late pregnancy because of potential harmful effects on fetal hemodynamics. It now appears that both PGE₂and nitric oxide are involved in maintaining ductal patency, but recent work indicates that endothelin and its type A receptor are involved in the actual contraction of the ductus arteriosus at birth.

Thus, corticosteroids may promote ductal closure, not only by inhibiting PGE₂synthesis, but also by their ability to stimulate endothelin-1 synthesis.

8. CONCLUSION

The foregoing selected examples of local action of eicosanoids were described to emphasize the diversity of function of these potent biologic agents. Further examples of the diversity of action of eicosanoids may be obtained by consulting the references provided.

Judging by the intense research efforts currently being extended in the eicosanoid field, it is clear that new information on the regulation and action of these biologically potent substances will continue to evolve.

Moreover, their important pathophysiologic role in certain diseases gives cause for optimism that the pharmacologic control of their synthesis and action will be of considerable therapeutic value.

ACKNOWLEDGMENTS

Thanks are due to my colleagues Drs. Robert C.

Skarnes and David Kupfer for critical evaluation of the text. The investigations of the author and his group that are included in this chapter were supported by National Institutes of Health grants HD-08129 and HD-20290, and US Department of Agriculture grants 98-35203- 6635 and 2004-35203-14176.

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