Triazolodiazepines are potent antagonists of platelet activating factor (PAF) in vitro and in vivo

Summary The triazolodiazepines brotizolam, triazolam and alprazolam inhibited PAF-induced human platelet aggregation in vitro (IC₅₀ = 0.54, 7.6 and 13.7 M, respectively) but showed only a weak or no effect against other aggregating agents (ADP, adrenaline, collagen, serotonin, arachidonic acid). In comparison, flunitrazepam and diazepam, two diazepines without the triazole ring, showed IC₅₀-values of 42 and 260 M, respectively. Flunitrazepam does not possess the specificity shown by the other compounds. Brotizolam and triazolam also inhibited PAF-induced human neutrophil aggregation in vitro, with IC₅₀-values 0.21 and 6.6 M, respectively.In anaesthetized guinea pigs, brotizolam (2.5 to 10 mg/kg p.o. or 0.1 to 0.5 mg/kg i.v.) or triazolam (20 to 100 mg/kg p.o.) inhibited dose-dependently the intrathoracic accumulation and aggregation of ¹¹¹Indium labelled platelets induced by an i. v. infusion of PAF (30 ng/kg × min).Brotizolam at doses of 1 to 10 mg/kg p. o. and 0.1 to 0.5 mg/kg i. v. inhibited dose-dependently the reduction in tidal volume (bronchoconstriction), the systemic hypotension and the lethal effect due to i. v. PAF in guinea pigs. Triazolam inhibited these effects of PAF at doses of 50 to 200 mg/kg p.o.PAF-induced systemic hypotension in rats can be reversed by cumulative i. v. doses (0.05 to 1.0 mg/kg) of brotizolam.In conclusion, these results show that triazolodiazepines, like brotizolam and triazolam, are potent inhibitors of PAF-induced effects in vitro and in vivo. Send offprint requests to J. Casals-Stenzel at the above address     

Endothelium-dependent vasodilator effects of platelet activating factor on rat resistance vessels.

1. To elucidate the mechanisms of the powerful and long-lasting hypotension produced by platelet activating factor (PAF), its effects on perfusion pressure in the perfused mesenteric arterial bed of the rat were examined. 2. Infusion of PAF (10(-11) to 3 x 10(-10) M; EC50 = 4.0 x 10(-11) M; 95%CL = 1.6 x 10(-11) - 9.4 x 10(-11) M) and acetylcholine (ACh) (10(-10) to 10(-6) M; EC50 = 3.0 +/- 0.1 x 10(-9) M) produced marked concentration-dependent vasodilatations which were significantly inhibited by treatment with detergents (0.1% Triton X-100 for 30 s or 0.3% CHAPS for 90 s). 3. Pretreatment with CV-6209, a PAF antagonist, inhibited PAF- but not ACh-induced vasodilation. 4. Treatment with indomethacin (10(-6) M) had no effect on PAF- or ACh-induced vasodilatation. 5. These results demonstrate that extremely low concentrations of PAF produce vasodilatation of resistance vessels through the release of endothelium-derived relaxing factor (EDRF). This may account for the strong hypotension produced by PAF in vivo.     

Electrophysiological and arrhythmogenic effects of platelet activating factor during normal perfusion, myocardial ischaemia and reperfusion in the guinea-pig.

1. Platelet activating factor (PAF) is often used to study the effects of platelet activation. While direct myocardial electrophysiological effects of PAF have been described in superfused myocardial tissue, little is known about its actions on the whole heart. 2. The cellular electrophysiological and arrhythmogenic effects of PAF (10(-11)M, 10(-10)M and 10(-9)M) were studied during normal perfusion, global myocardial ischaemia and reperfusion in Langendorff-perfused guinea-pig hearts at 32 degrees C. 3. PAF (10(-9)M) increased the incidence of ventricular fibrillation during ischaemia and reduced action potential duration (APD) during normal perfusion and early myocardial ischaemia (10(-9)M and 10(-10)M). PAF also reduced refractory period (RP) during normal perfusion (10(-9)M) and early ischaemia (10(-9)M and 10(-10)M). PAF prevented recovery of APD (10(-9)M) and RP (10(-9)M and 10(-10)M) during reperfusion. PAF at a concentration of 10(-11)M had no electrophysiological effects. 4. PAF (10(-9)M) increased the QRS width of the electrocardiogram during late ischaemia while 10(-10)M PAF raised pacing threshold during late ischaemia. 5. Perfusion pressure was increased, and developed tension decreased by 10(-9)M PAF. 6. These results demonstrate that PAF has direct myocardial electrophysiological effects in the whole heart which occur during normal perfusion and are capable of augmenting the effects of myocardial ischaemia, but are independent of the presence of platelets.     

Preparation of 7-substituted ginkgolide derivatives: potent platelet activating factor (PAF) receptor antagonists

Ginkgolides are structurally unique constituents of Ginkgo biloba extracts and are known antagonists of the platelet-activating factor (PAF) receptor. Ginkgolide C is 25-fold less potent than ginkgolide B as a PAF receptor antagonist, due to the presence of the 7beta-OH. Recently, we found that 7alpha-fluoro ginkgolide B was equipotent to ginkgolide B underlining the critical importance of the 7-position of ginkgolides for PAF receptor activity. Herein we describe the synthesis of a series of ginkgolide B derivatives with modifications at the 7-position and the pharmacological evaluation of these derivatives as assayed by cloned PAF receptors. In two cases nucleophilic attack on a 7beta-O-triflate ginkgolide B did not lead to the expected products, but gave rise to two unprecedented ginkgolide derivatives, one with a novel rearranged skeleton. Furthermore, standard reduction of 7alpha-azido ginkgolide B did not give the expected primary amine, but instead yielded alkylated amines depending on the solvent employed. Pharmacological testing with cloned PAF receptors showed that ginkgolides with 7alpha-substitutents had increased affinity compared to 7beta-substituents, in particular 7alpha-chloro ginkgolide B, the most potent nonaromatic ginkgolide derivative described to date with a K(i) value of 110 nM.     

Pharmacological modulation of platelet activating factor (PAF)-induced bronchoconstriction and hypertension in anaesthetized guinea-pigs

The effect of PAF has been examined in anaesthetized guinea-pigs. Intravenous (i.v.) administration of PAF (10 ng kg-1) did not modify the respiratory response but decreased the arterial blood pressure. A high dose of PAF (200 ng kg-1) caused marked bronchoconstriction and concomitant hypertension. The cyclooxygenase inhibitors aspirin (5 mg kg-1) and indomethacin (5 mg kg-1) and the thromboxane A2 (TXA2) receptor antagonist BM-13.177 (1 mg kg-1) failed to inhibit the peak bronchoconstrictive response but significantly inhibited the prolonged response following peak response. These inhibitors also attenuated PAF-induced hypertension. On the other hand, the lipoxygenase inhibitors phenidone (10 mg kg-1) and NDGA (5 mg kg-1) and the leukotriene (LT) receptor antagonist FPL-55712 (2 mg kg-1) affected neither bronchoconstriction nor hypertension induced by PAF. However, when aspirin was given in combination with NDGA, phenidone or FPL-55712, the peak and the following prolonged bronchoconstriction were significantly inhibited. The suppression of PAF-induced hypertension by aspirin was not further inhibited by the combination of these inhibitors. These results indicate that in anaesthetized guinea-pigs PAF-induced bronchoconstriction is composed of a dual response, a direct action (peak response) and an indirect action (prolonged response). The latter may be produced by the generation of TXA2 and lipoxygenase products, while PAF-induced hypertension is indirectly mediated by the generation of TXA2.     

Endothelium-dependent relaxant action of platelet activating factor in the rat mesenteric artery

Summary Vasorelaxant action of platelet activating factor (PAF) was examined in perfused mesenteric vascular beds and mesenteric artery strips isolated from rats. PAF caused a dose-dependent vasodilation of norepinephrine-contracted mesenteric vascular bed, which was sensitive to CV-3988, a PAF antagonist, but insensitive to tetrodotoxin, atropine, propranolol and indomethacin. PAF also caused a relaxation of phenylephrine-contracted mesenteric artery strips at above 3 × 10^–12 M. Much higher concentrations of PAF were required to relax the aorta, carotid and pulmonary arteries. The PAF- and acetylcholine (ACh)-induced relaxations of mesenteric artery were dependent on the presence of endothelium and were inhibited by either hydroquinone and methylene blue, which inhibit the action of endothelium-derived relaxing factor (EDRF), or L-canavanine, which inhibits the formation of nitric oxide from l-arginine. Phospholipase AZ inhibitors such as quinacrine and ONORS-082 abolished the relaxation induced by ACh but did not affect that by PAF. Thus, PAF induces a vasorelaxation by releasing EDRF from endothelial cells as ACh does, although the pathway to produce the substances by PAF may be different from that by ACh. Send offprint requests to K. Ito at the above address     

Platelet hyperaggregability to platelet activating factor (PAF) in non-ST elevation acute coronary syndromes

It is known that myocardial ischaemia increases platelet aggregatory response to various agonists, ex vivo. We investigated the platelet aggregatory response to platelet activating factor (PAF), ex vivo, in patients with non-ST elevation acute coronary syndromes and determined the specificity and sensitivity of this response. Thirty-two consecutive patients with non-ST elevation acute coronary syndromes and 20 healthy volunteers were studied. Platelet aggregation in platelet-rich plasma was studied on the day of admission. The maximal aggregation achieved within 2 min after the addition of PAF (100 nM) was expressed as a percentage of 100% light transmission. PAF EC50 values were defined as the concentration that induces 50% of maximal aggregation. The PAF EC50 values of the non-ST elevation acute coronary syndromes patients were significantly lower compared to those of the controls (p < 0.0001). The maximal percentage of aggregation was also significantly higher (p < 0.0005). Ninety-one per cent of the patients were correctly classified using PAF EC50 values (specificity 90.0% and sensitivity 91.2%); the corresponding results using the maximal percentage of aggregation were 80% (specificity 70.0% and sensitivity 87.5%). The estimated values used as thresholds were 22.47 nM and 17.97 for the PAF EC50 and the maximal percentage of aggregation, respectively. The results of the present study suggest that platelet hyperaggregability to PAF, ex vivo, in non-ST elevation acute coronary syndromes is characterised by a high specificity and sensitivity, and thus it may represent a mechanism contributing to the pathophysiology of acute coronary syndromes.     

Generation of platelet activating factor (PAF) by a human lung epithelial cell line

A human lung epithelial cell line (ATC-CCL-185) was cultured in nutrient Ham-F12 medium. Cells in monolayers were stimulated with either ionophore A23187 (1 microM) or phorbol myristate acetate (PMA, 0.2 microM) for various periods of time. Samples were analysed by HPLC and the presence of platelet activating factor (PAF) was detected by bioassay of the release of [3H]serotonin from rabbit platelets undergoing aggregation. The ATC-CCL 185 cells were found to synthesize PAF following activation with either PMA or ionophore. Ionophore at 1 microM was found to be more potent than PMA at 0.2 microM in the induction of PAF synthesis (congruent to 80 ng/mg protein). The synthesis of PAF through ionophore stimulation reached a maximum at 5 min, whereas PMA stimulation peaked at 15-20 min. PMA induced approximately one third the level of PAF synthesis by the ionophore. The PAF synthesized by these CCL185 cells was found to be mainly associated with the cell membrane with less than 10% released into the medium. Release of PAF into cell supernatant was dependent on the presence of bovine serum albumin (BSA). In the absence of BSA, a large portion (approximately 90%) of PAF was found to be cell associated, and only 60% when BSA concentration reached greater than or equal to 0.2%. These results demonstrate the ability of this lung epithelial cell line to synthesis PAF thus, suggesting that epithelial cells might participate in the process of inflammatory lung diseases, through the generation of this important mediator.     

Study of platelet aggregation induced by platelet activating factor (PAF) after administration of ticlopidine or aspirin

Platelet aggregation induced by platelet activating factor (PAF) was studied in 95 subjects: 39 controls, 23 patients receiving aspirin and 33 receiving ticlopidine. Potentiation of aggregation by concentrations of adrenaline unable to induce aggregation when used alone was also assessed. The 33 patients treated with ticlopidine showed a highly significant fall of platelet aggregation (p less than 0.001) at the three concentrations of PAF used. The 23 subjects receiving aspirin showed a diminution of platelet aggregation induced by PAF due to inhibition of ADP release. In these last two groups, adrenaline often potentiated platelet aggregation. However, this phenomenon was absent in subjects having taken aspirin in the hours before blood was drawn. This study demonstrates ticlopidine's inhibitory action on PAF-induced aggregation and confirms ticlopidine's role in reducing platelet aggregation by ADP, which has previously been demonstrated.     

Contribution of ADP to the amplification of primary platelet aggregation by platelet activating factor (PAF): modulatory role of aspirin

Platelet Activating Factor (PAF)-induced human platelet aggregation in citrated plasma is accompanied by activation of the cyclo-oxygenase pathway and release of intracellular constituents including Adenosine-5'-diphosphate (ADP). Inhibition of the cyclo-oxygenase pathway by aspirin prevented the amplification of primary platelet aggregation induced by threshold concentrations of PAF. Removal of ADP by enzymatic systems had little or no effect on PAF-induced full aggregation, but reversed the aggregating effect of PAF (at 10 times threshold concentrations) on 'aspirinated' platelets. Aspirin also prevented the synergism between PAF and ADP when subthreshold concentrations of both compounds were combined. Similar results were obtained in heparinized platelet-rich plasma. Thus, ADP may amplify the primary response to PAF but its role is modulated by the availability of the cyclo-oxygenase pathway products.     

The role of cycloxygenase-2 in the rodent kidney following ischaemia/reperfusion injury in vivo

The role of cyclooxygenase-2 (COX-2) in the pathophysiology of renal ischaemia/reperfusion injury is still not fully understood. In order to elucidate the role of COX-2 in ischaemia/reperfusion injury of the kidney, we have evaluated the effects of ischaemia/reperfusion on renal dysfunction and injury in (i) rats treated with either vehicle or the selective COX-2 inhibitor parecoxib, and (ii) wild-type mice or mice in which the gene for COX-2 has been deleted (COX-2 knock-out mice or COX-2(-/-)). Rats were subjected to bilateral renal ischaemia (45 min) and reperfusion (6 h), and received parecoxib (20 mg/kg, i.v.) 30 min prior to ischaemia and 3 h after the commencement of reperfusion. Serum urea, serum creatinine, serum aspartate aminotransferase, creatinine clearance and fractional excretion of sodium were all used as indicators of renal dysfunction and injury. Mice (wild-type and COX-2(-/-)) were subjected to bilateral renal ischaemia (30 min) and reperfusion (24 h) after which renal dysfunction (serum urea and creatinine) and renal injury was assessed by histological analysis. Parecoxib significantly augmented the degree of renal dysfunction and injury caused by ischaemia/reperfusion in the rat. In addition, the degree of renal injury and dysfunction caused by ischaemia/reperfusion was also significantly augmented in COX-2(-/-) mice when compared to their wild-type littermates. These findings support the view that metabolites of COX-2 protect the kidney against ischaemia/reperfusion injury, and (ii) that selective inhibitors of COX-2 may worsen renal dysfunction and injury in conditions associated with renal ischaemia.     

Glomerular effects of cholera toxin in isolated perfused rat kidney: a potential role for platelet activating factor.

Cholera toxin (MW 84 kDa) is now considered a pharmacological tool to study the adenylyl cyclase system and a stimulus to generate platelet activating factor in the intestinal tract. We used this toxin to evaluate the renal haemodynamics, glomerular filtration function, tubular sodium transport and toxicity in isolated perfused rat kidney. Kidneys from adult male Wistar rats were isolated for perfusion. The perfusion fluid was modified Krebs-Henseleit solution and the samples were analyzed for sodium, potassium, inulin and osmolality. Clearance techniques were used to calculate physiological parameters. Cholera toxin (1.0 microg/ml) caused a significant time-dependent reduction of glomerular filtration rate and urinary flow. This toxin also caused a small, but consistent reduction in fractional proximal sodium reabsortion (toxin = 67.43+/-2.42% versus control = 79.26+/-5.80%; P<0.025). WEB 2086, a platelet activating factor receptor antagonist at 100 microg/ml completely blocked the effects induced by cholera toxin on glomerular filtration rate, fractional proximal sodium reabsortion and urinary flow. In contrast to cholera toxin, dibutyryl-cyclic AMP (10(-5) M) significantly increased glomerular filtration rate (Db-cyclic AMP = 0.651+/-0.035 versus control = 0.514+/-0.043 ml x g(-1) x min(-1); P<0.025) in isolated perfused kidneys. Db-cyclic AMP caused a similar, but more severe reduction in fractional proximal sodium reabsortion (Db-cyclic AMP = 54.21+/-2.35% versus control = 70.10+/-3.24%; P<0.025). In addition Db-cyclic AMP increased significantly the urinary flow (Db-Cyclic AMP = 0.290+/-0.018 versus control = 0.179+/-0.026 ml x g(-1) x min.(-1); P<0.025). WEB 2086+ Db-cyclic AMP also caused a significant increase in the urinary flow with maximal effect at 90 min. (WEB+Db-cyclic AMP = 0.26+/-0.01 versus control = 0.15+/-0.01 ml x g(-1) x min.(-1); n = 8, P<0.025). Cholera toxin caused a decrease of urinary flow (toxin = 0.034+/-0.004 versus control = 0.145+/-0.02 ml x g(-1) x min.(-1); P<0.025), this effect was also completely abolished by WEB 2086 when it was injected previously to toxin. When only WEB 2086 was injected, the functional parameters remained stable throughout the perfusion time. Cholera toxin had no effect on renal vascular resistance, renal perfusate flow or tissue potassium, suggesting renal integrity in kidneys treated with this toxin. The results suggest that cholera toxin effects in the perfused rat kidney are primarily mediated by platelet activating factor.     

Protective effects of a platelet activating factor (PAF) antagonist and its combined treatment with prostaglandin (PG) E1 in traumatic shock

We have investigated the role of platelet activating factor (PAF) in the pathogenesis of a murine model of traumatic shock using CV-6209, a specific antagonist of PAF. CV-6209, at a dose of 1 mg/kg (i.v.) given after trauma, significantly improved survival rate at 150 min and overall survival time. Furthermore, the plasma accumulation of the lysosomal hydrolase, cathepsin D, and a cardiotoxic peptide, myocardial depressant factor (MDF), were also attenuated by CV-6209 in traumatic shock. Combined treatment employing low doses of CV-6209 [0.2 mg/kg, i.v. and prostaglandin (PG) E1, 0.8 microgram/kg/min] in this shock model was also examined. CV-6209 (0.2 mg/kg) or PGE1 (0.8 microgram/kg/min) alone at these doses showed only minimal effects on survival, or plasma cathepsin D or MDF activities. However, combined treatment with CV-6209 (0.2 mg/kg, i.v.) and PGE1 (0.8 microgram/kg/min) significantly improved survival rate at 150 min, overall survival time, and decreased the accumulation of plasma MDF. These results suggest that PAF may play an important pathophysiologic role in traumatic shock in rats. Moreover, combination therapy using a PAF antagonist and PGE1 may be useful for the treatment of traumatic shock.     

Modulation of in vitro immune reactions by platelet activating factor and a platelet activating factor antagonist

Platelet activating factor (PAF) was found to suppress primary and secondary mixed lymphocyte reactions and BN52021, a naturally occurring PAF antagonist, blocked PAF-mediated suppression and enhanced the mixed lymphocyte reactions. The effect of delayed addition of PAF or BN52021 24 h or later after the initiation of cultures reduced the suppressive and enhancing effects, respectively. The removal of the antagonist BN52021 from mixed lymphocyte cultures up to 72 h after their initiation also was found to eliminate the potentiating effect of this antagonist. The continuous presence of PAF in mixed lymphocyte cultures used to generate cytotoxic lymphocytes suppressed the generation of effector cells while the addition of BN52021 elicited an enhanced level of cell-mediated cytotoxicity in such cultures. BN52021 enhanced the cytotoxic activity in such cultures irrespective of the presence of exogenous interleukin-2, suggesting that the antagonist-mediated enhancement is not due to the enhanced production of interleukin-2 by cells in the mixed cultures. The experiments reported here provide evidence for a role of PAF in modulating the complex interactions that take place in the initiation of cellular immune reactions. Furthermore, the results of these experiments indicate that the immunoregulatory action of PAF can be modulated by an antagonist that exerts its action independently of the production of the lymphocyte growth factor, interleukin-2.     

Role of platelet-activating factor (PAF) in platelet accumulation in rabbit skin: effect of the novel long-acting PAF antagonist, UK-74,505.

1. The contribution of platelet-activating factor (PAF) to platelet deposition and oedema formation induced by exogenous soluble mediators, zymosan particles and associated with a reversed passive Arthus (RPA) reaction in rabbit skin was investigated by use of a novel long-acting PAF receptor antagonist, UK-74,505. 2. Oedema formation and platelet accumulation were simultaneously measured by i.v. injection of [125I]-albumin and 111In-labelled rabbit platelets. UK-74,505 was either administered i.v. or used to pretreat radiolabelled platelets in vitro before their injection into recipient animals. Platelets pretreated with UK-74,505 were also labelled with the fluorescent calcium indicator, Fura-2, to assess their ex vivo reactivity to PAF at the end of the in vivo experiment. 3. UK-74,505 (0.5 mg kg-1), administered i.v., inhibited PAF-induced oedema formation, but did not affect oedema induced by zymosan particles, bradykinin (BK), histamine, formyl-methionyl-leucylphenylalanine (FMLP), zymosan-activated plasma (ZAP, as a source of C5a des Arg), leukotriene B4 (LTB4) or interleukin-8 (IL-8). 4. UK-74,505, administered i.v. also suppressed the small platelet accumulation induced by exogenous PAF, but had no effect on accumulation induced by IL-8 or ZAP. Although oedema induced by zymosan was not affected by i.v. UK-74,505, zymosan-induced platelet accumulation was significantly attenuated by the antagonist. 5. The RPA reaction in rabbit skin was associated with marked oedema formation and platelet accumulation which were both inhibited by i.v. UK-74,505.(ABSTRACT TRUNCATED AT 250 WORDS)     

An antagonistic activity of etizolam on platelet-activating factor (PAF). In vitro effects on platelet aggregation and PAF receptor binding

The antagonistic effect of etizolam, an anti-anxiety drug, on platelet-activating factor (PAF) was investigated in rabbit platelets in vitro. Etizolam inhibited PAF-induced aggregation in a dose-dependent manner, with an IC50 of 3.8 microM, about one tenth that of triazolam (IC50 = 30 microM). At 300 microM, it inhibited both ADP and arachidonic acid-induced aggregation only slightly, while the other anti-anxiety drugs tested had no effect on PAF-induced aggregation even at this concentration. Etizolam and triazolam inhibited the specific binding of 3H-PAF to PAF receptor sites on washed rabbit platelets with IC50 values of 22 nM and 320 nM, respectively. Diazepam and estazolam were inactive even at 1 microM. These results indicate that etizolam is a specific antagonist of PAF.     

Analogues of platelet activating factor (PAF). 2. Some modifications of the glycerine backbone

Racemic analogues of platelet activating factor (PAF) that contain a methylene group between the C2 and C3 carbon atoms (39) or between the C1 and C2 carbon atoms (40) have been synthesized. These compounds show reduced platelet aggregation and hypotensive activity as measured against racemic C16 PAF. Compounds in which the C1 carbon atom of PAF is substituted with one or two methyl groups (41 and 42, respectively) or the C3 carbon is substituted with a single methyl group (43) have been synthesized. Platelet aggregation and hypotensive responses produced by these compounds are significantly less than those obtained with racemic C16 PAF. None of the above compounds exhibit a separation of the platelet aggregation and hypotensive activities.     

Analogues of platelet activating factor (PAF). 1. Some modifications of the alkoxy chain

Analogues of platelet activating factor (PAF) in which the ether oxygen has been removed (6) and in which the alkoxy chain at C1 has been replaced with a o-, p-, or m-alkylphenoxy group (30, 31, and 35, respectively) have been prepared. Compound 6 shows reduced platelet aggregation and hypotensive activity in comparison with C16 and C18 PAF. The results obtained for compounds 30, 31, and 35 indicate that the hypotensive and platelet aggregating responses are sensitive to structural modification of the ether chain. The ortho analogue 30 shows no platelet aggregating activity and only a weak hypotensive response. The para analogue 31 exhibits a moderate decrease in activity in both assays. The meta analogue 35 is the most active of the three.     

粉防己碱对兔血小板聚集和血小板活化因子生成的影响(英文)

目的:探讨粉防己碱(Tet)对兔血小板聚集和PAF生成的影响.方法:卡西霉素(Cal)和PAF诱导血小板聚集的聚集率和Tet对血小板聚集的抑制率被测定;给予或未给予Tet处理之血小板用Cal刺激释放PAF的量也被测定.结果:在4—64 μmol·L~(-1)浓度范围,Tet明显抑制Cal和PAF诱导的血小板聚集.IC_(50)值分别为8.6μmol·L~(-1)和14.0μmol·L~(-1).Tet也浓度依赖性的抑制Cal诱导血小板释放PAF,IC_(50)值为21.0μmol·L~(-1).结论:Tet抑制血小板聚集作用与抑制内源性PAF生成有关.     

Platelet activating factor (PAF) is a potent stimulator of porcine tracheal fluid secretion in vitro

Mucus hypersecretion is a major clinical feature of chronic obstructive lung diseases such as asthma. The possible role of the inflammatory and bronchoconstrictor ether lipid PAF (platelet activating factor) has been studied in isolated porcine trachea with the tantalum 'hillock' technique used to visualize fluid production from tracheal submucosal glands. PAF caused a rapid, dose-dependent (0.001-1 nM) stimulation of fluid secretion which could be detected after 5 min and which increased with time up to at least 15 min. The PAF-induced fluid secretion was unaffected by both antagonists of histamine, acetylcholine and leukotriene D4 and inhibitors of prostaglandin and leukotrienes synthesis. A purported PAF receptor antagonist (CV 3988) inhibited the PAF responses in a dose-dependent manner implying a receptor-mediated event. These results may be of relevance to the mucus hypersecretion seen in chronic airway diseases.