What are prostaglandins / What does prostaglandin do in females? Prostaglandins, Leukotrienes (Eicosanoids) and Platelet Activating Factor /

 Prostaglandins, Leukotrienes (Eicosanoids) and Platelet Activating Factor :


Prostaglandins and Leukotrienes (Eicosanoids):

  Prostaglandins (PGs) and Leukotrienes (LTs) are biologically active derivatives of 20 carbon atom polyunsaturated essential fatty acids that are released from cell membrane phospholipids. They are the major lipid derived autacoids.

     In the 1930s human semen was found to contract isolated uterine and other smooth muscle strips and to cause fall in BP in animals. The active principle was termed ‘prostaglandin’, thinking that it was derived from prostate. Only in the 1960s it was shown to be a mixture of closely related compounds, the chemical structures were elucidated and widespread distribution in the body was revealed. In 1970s it became clear that aspirin like drugs act by inhibiting PG synthesis, and that in addition to the classical PGs (PGEs and PGFs), thromboxane (TX), prostacyclin (PGI) and leukotrienes (LTs) were of great biological importance. Bergstrom, Samuelsson and Vane got the Nobel prize in 1982 for their work on PGs and LTs. Over the past 40 years PGs and LTs have been among the most intensely investigated substances.

CHEMISTRY, BIOSYNTHESIS AND DEGRADATION :

   Chemically, PGs may be considered to be derivatives of prostanoic acid, though prostanoic acid does not naturally occur in the body. It has a five membered ring and two side chains projecting in opposite directions at right angle to the plane of the ring. There are many series of PGs and thromboxanes (TXs) designated A, B, C....I, depending on the ring structure and the substituents on it. Each series has members with subscript 1, 2, 3 indicating the number of double bonds in the side chains.

  Leukotrienes are so named because they were first obtained from leukocytes (leuko) and have 3 conjugated double bonds (triene). They have also been similarly designated A, B, C.....F and given subscripts 1, 2, 3, 4.


     In the body PGs, TXs and LTs are all derived from eicosa (referring to 20 C atoms) tri/tetra/penta enoic acids. Therefore, they can be collectively called eicosanoids. In human tissues, the fatty acid released from membrane lipids in largest quantity is 5,8,11,14 eicosa tetraenoic acid (arachidonic acid). During PG, TX and prostacyclin synthesis, 2 of the 4 double bonds of arachidonic acid get saturated in the process of cyclization, leaving 2 double bonds in the side chain. Thus, subscript 2 PGs are the most important in man, e.g. PGE2 , PGF2α, PGI2 , TXA2 . No cyclization or reduction of double bonds occurs during LT synthesis—the LTs of biological importance are LTB4 , LTC4 , LTD4 ..

 Eicosanoids are the most universally distributed autacoids in the body. Practically every cell and tissue is capable of synthesizing one or more types of PGs or LTs. As such, they have potent and a very wide range of biological activity. The pathways of biosynthesis of eicosanoids are summarized in Fig. 13.1.

  There are no preformed stores of PGs and LTs. They are synthesized locally and the rate of synthesis is governed by the rate of release of arachidonic acid from membrane lipids in  response to appropriate stimuli. These stimuli activate hydrolases, including phospholipase A, probably through increased intracellular Ca2+.


Cyclooxygenase (COX) pathway :

It generates eicosanoids with a ring structure (PGs, TXs, prostacyclin) while lipoxygenase (LOX) produces open chain compounds (LTs). All tissues have COX—can form cyclic endoperoxides PGG2 and PGH2 which are unstable compounds. Further course in a particular tissue depends on the type of isomerases or other enzymes present in it. PGE2 and PGF2α are the primary prostaglandins (name based on the separation procedure: PGE partitioned into Ether while PGF into phosphate [Fosfat in Swedish] buffer; α in PGF2α refers to orientation of OH group on the ring). PGs A, B and C are not found in the body: they are artifacts formed during extraction procedures. Lung and spleen can synthesize the whole range of COX products. Platelets primarily synthesize TXA2 which is—chemically unstable, spontaneously changes to TXB2 . Endothelium mainly generates prostacyclin (PGI2 ) which is also chemically unstable and rapidly converts to 6-keto PGF1α.

  Cyclooxygenase is known to exist in two isoforms COX-1 and COX-2. While both isoforms catalyse the same reactions, COX-1 is a constitutive enzyme in most cells—it is synthesized and is active in the basal state; the level of COX-1 activity is not much changed once the cell is fully grown. On the other hand, COX-2 normally present in insignificant amounts, is inducible by cytokines, growth factors and other stimuli during the inflammatory response. It is believed that eicosanoids produced by COX-1 participate in physiological (house keeping) functions such as secretion of mucus for protection of gastric mucosa, haemostasis and maintenance of renal function, while those produced by COX-2 lead to inflammatory and other pathological changes. However, certain sites in kidney, brain and the foetus constitutively express COX-2 which may play physiological role.

     A splice variant of COX-1 (designated COX-3) has been found in the dog brain. This isoenzyme is inhibited by paracetamol and is implicated in the genesis of fever, but the exact role in humans is not known.

Lipoxygenase pathway  :

     This pathway appears to operate mainly in the lung, WBC and platelets. Its most important products are the LTs, (generated by 5-LOX) particularly LTB4 (potent chemotactic) and LTC4 , LTD4 which together constitute the ‘slow reacting substance of anaphylaxis’ (SRS-A) described in 1938 to be released during anaphylaxis. A membrane associated transfer protein called FLAP (five lipoxygenase activating protein) carrys arachidonic acid to 5-LOX, and is essential for the synthesis of LTs. Platelets have only 12-LOX.
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  Apart from PGs and LTs, a number of other active products can be generated from arachidonic acid at certain sites. HPETEs produced by LOX can also be converted to hepoxilins, trioxilins and lipoxins. A third enzymatic pathway involving cytochrome P450 can metabolize arachidonic acid into 19- and 20-HETEs and epoxyeicosatrienoic acids. Free radicals can attack arachidonic acid to produce isoprostanes nonenzymatically. Brain cells couple arachidonic acid with ethanolamine to produce anandamide and a few other related eicosanoids which are now recognized to be the endogenous cannabinoid receptor ligands. Accordingly, they produce cannabis like effects. Like the other eicosanoids, they are synthesized only when needed at the site of action.

Inhibition of synthesis :

 Synthesis of COX products can be inhibited by nonsteroidal antiinflammatory drugs (NSAIDs). Aspirin acetylates COX at a serine residue and causes irreversible inhibition, while other NSAIDs are competitive and reversible inhibitors. Most NSAIDs are nonselective COX-1 and COX-2 inhibitors, but some later ones like celecoxib, etoricoxib are selective for COX-2.

    The sensitivity of COX in different tissues to inhibition by these drugs varies; selective inhibition of formation of certain products may be possible at lower doses. NSAIDs do not inhibit the production of LTs. Rather, LT production may be increased since all the arachidonic acid becomes available to the LOX pathway.
  Zileuton inhibits LOX and decreases the production of LTs. It was used briefly in asthma, but has been withdrawn.

 Glucocorticosteroids inhibit the release of arachidonic acid from membrane lipids (by enhancing production of proteins called annexins which inhibit phospholipase A2 ). Production of all eicosanoids—PGs, TXs and LTs is indirectly curtaited. Moreover, steroids inhibit the induction of COX-2 by cytokines at the site of inflammation.

Degradation :

Biotransformation of arachidonates occurs rapidly in most tissues, but fastest in the lungs. Most PGs, TXA2 and prostacyclin have plasma t½ of a few seconds to a few minutes. First a specific carrier mediated uptake into cells occurs, the side chains are then oxidized and double bonds are reduced in a stepwise manner to yield inactive metabolites. Metabolites are excreted in urine. PGI2 is catabolized mainly in the kidney.


ACTIONS AND PATHOPHYSIOLOGICAL ROLES :


Prostaglandins, thromboxanes and prostacyclin :

 The cyclic eicosanoids produce a wide variety of actions depending upon the particular PG (or TX or PGI), species on which tested, tissue, hormonal status and other factors. PGs differ in their potency to produce a given action and different PGs sometimes have opposite effects. Even the same PG may have opposite effects under different circumstances. The important actions of PGs and TXA2 are summarized in Table 13.1. Since virtually all cells and tissues are capable of forming one or more PGs, these autacoids have been implicated as mediators or modulators of a number of physiological processes and pathological states.

1. CVS :

 PGE2 causes vasodilatation in most, but not all, vascular beds. PGF2α constricts many larger veins including pulmonary vein and artery. Fall in BP occurs when PGE2 is injected i.v., but PGF2α has little effect on BP.

  • PGI2 is uniformly vasodilatory and is more potent hypotensive than PGE2 . 
  • TXA2 consistently produces vasoconstriction. 
  • PGE2 and F2α stimulate heart by weak direct but more prominent reflex action due to fall in BP. The cardiac output increases.

 Role :

  • PGs do not circulate in blood and have no role in regulating systemic vascular resistance. However, PGI2 generated in the vascular endothelium, mainly by COX-2, appears to be involved in the regulation of local vascular tone as a dilator. 
  • PGE2 is continuously produced locally in the ductus arteriosus by COX-2 during foetal life and keeps it patent. At birth its synthesis stops and closure occurs. Aspirin and indomethacin induce closure when it fails to occur spontaneously. 
  • PGs, generated mainly by COX-2, along with LTs and other autacoids may mediate vasodilatation and exudation at the site of inflammation.
   

2. Platelets :

  TXA2 , which can be produced locally by platelets, is a potent inducer of aggregation and release reaction. On the other hand PGI2 (generated by vascular endothelium) is a potent inhibitor of platelet aggregation. PGE2 has dose dependent pro- and anti-aggregatory effects.

Role  :

 TXA2 produced by platelets and PGI2 produced by vascular endothelium probably constitute a mutually antagonistic system: preventing aggregation of platelets while in circulation and inducing aggregation on injury, when plugging and thrombosis are needed.

 Aspirin interferes with haemostasis by inhibiting platelet aggregation. TXA2 produced by platelet COX-1 plays an important role in amplifying aggregation. Before it is deacetylated in liver, aspirin acetylates COX-1 in platelets while they are in portal circulation. Further, platelets are unable to regenerate fresh COX-1 (lack nucleus: do not synthesize protein), while vessel wall is able to do so (fresh enzyme is synthesized within hours). Thus, at low doses, aspirin selectively inhibits TXA2 production and has antithrombotic effect lasting > 3 days.

3. Uterus :

  PGE2 and PGF2α consistently contract human uterus in vivo, both pregnant as well as nonpregnant. The sensitivity is higher during pregnancy and there is progressive modest increase with the advance of pregnancy. However, even during early stages, uterus is quite sensitive to PGs though not to oxytocin. PGs increase basal tone as well as amplitude of uterine contractions.

                          At term, PGs soften the cervix at low doses and make it more compliant.

Role :

  • Foetal tissues produce PGs. At term PGF2α has been detected in maternal blood. It is postulated that PGs mediate initiation and progression of labour. Aspirin has been found to delay the initiation of labour and also prolong its duration. 
  • Dysmenorrhoea in many women is associated with increased PG synthesis by the endometrium. This apparently induces uncoordinated uterine contractions which compress blood vessels causing uterine ischaemia and pain. Aspirin group of drugs are highly effective in relieving dysmenorrhoea in most women.

4. Bronchial muscle :

                                           PGF2α, PGD2 and TXA2 are potent bronchoconstrictors (more potent than histamine) while PGE2 is a powerful bronchodilator. PGI2 produces mild dilatation. Asthmatics are more sensitive to constrictor as well as dilator effects of PGs. PGE2 and PGI2 also inhibit histamine release and are effective by aerosol. However, these antiasthmatic effects of PGE2 and PGI2 cannot be exploited clinically because they produce irritation of the respiratory tract and have brief action.

Role :

 Asthma may be due to an imbalance between constrictor PGs (F2α, PGD2 , TXA2 ) and cysteinyl LTs on one hand and dilator ones (PGE2 , PGI2 ) on the other. In few individuals aspirin-like drugs consistently induce asthma, possibly by diverting arachidonic acid to produce excess LTC4 and D4 . This sensitivity is not shared by selective COX-2 inhibitors, indicating that suppression of COX-1 at the pulmonary site is responsible for the reaction. In allergic human asthma, LTs play a more important role, and COX inhibitors are without any effect in most patients.



 

5. GIT :

                 (i) In isolated preparations, the longitudinal muscle of gut is contracted by PGE2 and PGF2α while the circular muscle is either contracted (usually by PGF2α) or relaxed (usually by PGE2 ). Propulsive activity is enhanced in man, especially by PGE2 . This can cause colic and watery diarrhoea, which are important side effects. PGE2 acts directly on the intestinal mucosa and increases water, electrolyte and mucus secretion.

Role :

 PGs may be involved in mediating toxin induced increased fluid movement in secretory diarrhoeas. In certain diarrhoeas, aspirin can reduce stool volume, but it is not uniformly effective. PGs appear to play a role in the growth of colonic polyps and cancer. Association of lower incidence of colon cancer with regular intake of aspirin is now established. NSAIDs afford relief in familial colonic polyposis by reducing polyp formation.

(ii) PGE2 markedly reduces acid secretion in the stomach. Volume of juice and pepsin content are also decreased. It inhibits fasting as well as stimulated secretion. Release of gastrin is suppressed (see Fig. 47.1). The gastric pH may rise upto 7.0. PGI2 also reduces gastric secretion, but is less potent. Secretion of mucus and HCO3 ¯ by gastric mucosal epithelial cells is increased, as is mucosal blood flow. Thus, PGs are antiulcerogenic. 

Role

    PGs (especially PGI2 ) appear to be involved in the regulation of gastric mucosal blood flow. They may be functioning as natural ulcer protectives by enhancing gastric mucus and HCO3 ¯ production, as well as by improving mucosal circulation and health. The ulcerogenic action of NSAIDs appears to be due to loss of this protective influence.

Normally, gastric mucosal PGs are produced by COX­1. Selective COX-2 inhibitors are less ulcerogenic. However, COX-2 gets induced during ulcer healing, and COX-2 inhibitors have the potential to delay healing.

6. Kidney :

        PGE2 and PGI2 increase water, Na+ and K+ excretion and have a diuretic effect. PGE2 has a furosemide-like inhibitory effect on Cl¯ reabsorption as well. They cause renal vasodilatation and inhibit tubular reabsorption. PGE2 attenuates ADH action, and this adds to the diuretic effect. In contrast, TXA2 causes renal vasoconstriction. PGI2 , PGE2 and PGD2 evoke release of renin.

Role :

  •         PGE2 and PGI2 produced mainly by COX­2 in the kidney appear to function as intrarenal regulators of blood flow as well as tubular reabsorption in kidney. Accordingly, the NSAIDs, including selective COX-2 inhibitors, tend to retain salt and water. The diuretic action of furosemide is blunted by indomethacin—indicating a facilitatory role of PGs by increasing renal blood flow and/or augmenting inhibition of tubular reabsorption. 
  • Renin release in response to sympathetic stimulation, low salt intake and other influences may be facilitated by PGs.

7. CNS :

     PGs injected i.v. penetrate brain poorly, so that central actions are not prominent. However, injected intracerebroventricularly PGE2 produces a variety of effects—sedation, rigidity, behavioural changes and marked rise in body temperature. PGI2 also induces fever, but TXA2 is not pyrogenic.

Role :

  • PGE2 may mediate pyrogen induced fever and malaise. Aspirin and other inhibitors of PG synthesis are antipyretic. Pyrogens, including cytokines, released during bacterial infection trigger synthesis of PGE2 in the hypothalamus which resets the thermostat to cause fever. COX-2 is the major isoenzyme involved; selective COX-2 inhibitors are equally efficacious antipyretics. A role of COX-3 has also been proposed.
  • PGs may be functioning as neuromodulators in the brain by regulating neuronal excitability. A role in pain perception, sleep and some other functions has been suggested.

8. Sympathetic nerves :

   Depending on the PG, species and tissue, both inhibition as well as augmentation of NA release from sympathetic nerve endings has been observed. However, PGE2 mostly inhibits NA release.

Role  : 

           PGs may modulate sympathetic neurotransmission in the periphery.


9. Peripheral nerves :

    PGs (especially E2 and I2 ) sensitize afferent nerve endings to pain inducing chemical and mechanical stimuli (Fig. 13.2). They irritate mucous membranes and produce long lasting dull pain on intradermal injection.

Role :

          PGs serve as algesic agents during inflammation. They cause tenderness and amplify the action of other algesics. Inhibition of PG synthesis is a major antiinflammatory mechanism. Aspirin injected locally decreases pain produced by injection of bradykinin at the same site. Moreover, PGs released in the dorsal horn of spinal cord by painful stimuli sensitise the neurones to pain perception.




 10. Eye :
                       PGF2α induces ocular inflammation and lowers i.o.t by enhancing uveoscleral and trabecular outflow. Nonirritating congeners like latanoprost are now first line drugs in wide angle glaucoma (see p. 169). 

Role :                                                                                                                                            
               Locally produced PGs appear to facilitate aqueous humor drainage. The finding that COX-2 expression in the ciliary body is deficient in wide angle glaucoma patients supports this contention.

11. Endocrine system :


                                       PGE2 facilitates the release of anterior pituitary hormones (growth hormone, prolactin, ACTH, FSH and LH) as well as that of insulin and adrenal steroids. It has a TSH-like effect on the thyroid.

12. Metabolism :

                                 PGEs are antilipolytic, exert an insulin like effect on carbohydrate metabolism and mobilize Ca2+ from bone. They may facilitate hypercalcaemia due to bony metastasis.

 




    
    












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