Dietary fatty acids in human thrombosis and hemostasis.

The effects of fatty acids on hemostasis are controversial. It has been difficult to show convincing effects of saturated or monounsaturated fatty acids that are clearly related to hemostatic variables in humans. Unsaturated fatty acids alter platelet aggregation and processes related to coagulation and fibrinolysis. Indirect evidence exists that n-6 polyunsaturated fatty acids may exert favorable effects on thrombotic processes in vivo, but large clinical trials have failed to show benefits of 5-6 g linoleic acid (18:2n-6) or linolenic acid (18:3n-3)/d. Only long-chain n-3 fatty acids prolong the template bleeding time, and they may exert some beneficial effect on erythrocyte flexibility. It appears unlikely that n-3 fatty acids lower fibrinogen or interact with the fibrinolytic system directly. One prospective secondary prevention trial showed benefits that may have resulted from either an improved hemostatic profile or an antiarrhythmic effect. A similar time course of clinical improvement was noted with reduced rates of cardiac mortality and postoperative thrombosis in Norway during World War II, and this was associated with a drastic dietary alteration involving increased consumption of n-3 fatty acids and reduced consumption of saturated fatty acids. Further work is needed to develop better tools to examine in vivo hemostasis so that the mechanisms and eventual clinical utility of n-3 fatty acids can be elucidated in well-designed clinical trials.

Pentoxifylline does not alter the response to inhaled grain dust.

Pentoxifylline (PTX) has been shown to reduce sepsis-induced neutrophil sequestration in the lung and inhibit endotoxin-mediated release of tumor necrosis factor-alpha (TNF-alpha). Previously, we have shown that endotoxin appears to be the principal agent in grain dust causing airway inflammation and airflow obstruction following grain dust inhalation. To determine whether PTX affects the physiologic and inflammatory events following acute grain dust inhalation, 10 healthy, nonsmoking subjects with normal airway reactivity were treated with PTX or placebo (PL) followed by corn dust extract (CDE) inhalation (0.08 mL/kg), using a single-blinded, crossover design. Subjects received PTX (1,200 mg/d) or PL for 4 days prior to CDE inhalation and 400 mg PTX or PL on the exposure day. Both respiratory symptoms and declines in FEV1 and FVC occurred following CDE exposure in both groups, but there were no significant differences in the frequency of symptoms or percent declines from baseline in the FEV1 and FVC at any of the time points measured in the study. Elevations in peripheral blood leukocyte and neutrophil concentrations and BAL total cell, neutrophil, TNF-alpha, and interleukin-8 concentrations were measured 4 h following exposure to CDE in both the PTX- and PL-treated subjects, but no significant differences were found between treatment groups. These results suggest that pretreatment with PTX prior to inhalation of CDE, in the doses used in this study, does not alter the acute physiologic or inflammatory events following exposure to inhaled CDE.

n-3 fatty acids and human hypertension.

Clinical studies of the antihypertensive effects of n-3 fatty acids in different groups of patients are summarized, and recent information of possible mechanisms of antihypertensive effects is discussed. Because of marked differences between fatty acid metabolism and blood pressure control of humans and laboratory animals, the focus is on human studies.

Omega-3 fatty acids in respiratory diseases: a review.

The health benefits of dietary omega-3 fatty acids have been investigated in a variety of conditions but there have been few studies of their effects in human respiratory diseases. Although many of the physiological changes associated with omega-3 polyunsaturate ingestion have been attributed to alterations in endogenous eicosanoid production, effects on blood rheology, host-microbial interactions and lung surfactant production have also been described. In reviewing the literature, there is little evidence that these polyunsaturates have beneficial effects in allergic disorders, but they may have potential as modulators of respiratory diseases involving chronic inflammatory and infectious processes or impaired pulmonary blood flow. Further work on the effects of omega-3 fatty acids in several chronic pulmonary syndromes, for which there are currently no effective therapies, appears to be warranted.

Asymmetric incorporation of dietary n-3 fatty acids into membrane aminophospholipids of human erythrocytes.

Dietary supplementation with different classes of polyunsaturate fatty acids is known to result in their incorporation into cell membranes, but the effects of this on eicosanoid formation and other cell functions frequently does not correspond to the degree of alteration in total membrane fatty acids. This phenomenon may be related to the compartmentalization of polyunsaturate fatty acids both within the organelles and within membranes. Aminophospholipids are asymmetrically distributed across the membrane bilayers of most human cells. These phospholipids are highly enriched in polyunsaturated fatty acids, and are known to have specific interactions with a number of membrane proteins. To determine whether dietary n-3 fatty acids are preferentially incorporated into membrane lipids in a particular spatial pattern, we have utilized the nonpermeant aminophospholipid probe, trinitrobenzenesulfonic acid, to study the transmembrane molecular species distribution of human erythrocyte ethanolamine phospholipids and phosphatidylserines before and at the end of 4 weeks of dietary supplementation with n-3 fatty acids. Selective incorporation of n-3 fatty acids occurred in the inner membrane leaflet ethanolamine phospholipids, particularly into the alkenyl-acyl species. The n-3 species in phosphatidylserines, particularly 18:0 and 22:6 n-3 (sn-1 and sn-2, respectively), replaced n-6 and n-9 species. These data may provide a basis for different cell responses to n-3 fatty acid enrichment, and for different degrees of diet-induced alteration in responses involving inner and outer membrane leaflet functions.

The effect of linoleic, arachidonic and eicosapentaenoic acid supplementation on prostacyclin production in rats.

We examined the effect of dietary supplementation of linoleic acid (LA), arachidonic acid (AA) or eicosapentaenoic acid (EPA) to rats fed a diet low in linoleic acid on in vitro and in vivo production of prostacyclin. Male Sprague Dawley rats were fed a high-fat diet (50% energy as fat, 1.5% linoleic acid) for two weeks. Three of the groups were then supplemented orally with either 90 mg/d of LA, AA or EPA, all as the ethyl esters, for a further two weeks while remaining on the high-fat diet. Forty-eight hour urine samples were collected at the end of the second and fourth weeks. In vivo prostacyclin production was determined by a stable isotope dilution, gas chromatography/mass spectrometry assay for the major urinary metabolite of prostacyclins (2,3-dinor-6-keto-PGF1 alpha or PGI2-M and delta 17-2,3-dinor-6-keto-PGF1 alpha or PGI3-M). In vitro prostacyclin production was determined by radioimmunoassay of the stable metabolite (6-keto-PGF 1 alpha) following incubation of arterial tissue. Oral supplementation with AA resulted in a rise in plasma and aorta 20:4n-6, and increased in vitro prostacyclin and urinary PGI2-M production. EPA supplementation resulted in a rise in plasma and aorta 20:5n-3 and 22:5n-3, and a decline in plasma 20:4n-6, but not in the aorta. In the EPA-supplemented group, the in vitro prostacyclin and the urinary PGI3-M increased, but urinary PGI2-M decreased. The increase in in vitro prostacyclin production in the EPA-supplemented rats was unexpected and without obvious explanation.(ABSTRACT TRUNCATED AT 250 WORDS)

Suppression of eicosanoid biosynthesis during coronary angioplasty by fish oil and aspirin.

BACKGROUND: Percutaneous transluminal coronary angioplasty (PTCA) is an acute, localized stimulus to platelet and vascular function. Periprocedural cardiovascular complications are reduced by moderate-dose aspirin (ASA), presumably due to inhibition of thromboxane (TX) A2. METHODS AND RESULTS: Excretion of TXA2 and prostacyclin (PGI2) metabolites in urine increased during PTCA. Pretreatment for 3 days with either moderate- (325 mg/day) or low-dose (80 mg/day) ASA inhibited the increase in both eicosanoids. Pretreatment for 3 weeks with fish oil (10 g/day) only partially suppressed TXA2. Formation of trienoic eicosanoids and accumulation of omega-3 fatty acids in platelet membranes confirmed fish oil ingestion. Although basal PGI2 was not inhibited, the PTCA-related increment was suppressed. CONCLUSIONS: PTCA results in an acute, transient alteration of eicosanoid biosynthesis consistent with accelerated platelet-vascular interactions. Pretreatment for 3 days with moderate or low doses of ASA suppresses TXA to a similar extent during PTCA, and their effects on acute cardiovascular complications of this procedure are likely to be comparable. It is unlikely that even prolonged pretreatment with fish oil can substitute for the platelet inhibitory action of ASA during PTCA. Suppression of PGI2 may contribute to the residual acute periprocedural complication rate in patients taking ASA.

Urinary excretion of diols derived from eicosapentaenoic acid during n-3 fatty acid ingestion by man.

Epoxides and fatty acid diols derived from arachidonate by the action of cytochrome P-450 appear in human urine and have biological activities. Dietary eicosapentaenoic acid gives rise to prostaglandins in vivo, but vascular effects of n-3 supplements do not all correlate with altered types or amounts of in vivo cyclooxygenase products. We investigated whether dietary eicosapentaenoic acid could also be metabolized by cytochrome P-450, by assessing the excretion of its vicinal diols. Utilizing gas chromatography/negative chemical ionization mass spectrometry, we have found that humans ingesting n-3 fatty acids excrete vicinal diols of eicosapentaenoic acid in substantial quantities.

Polyunsaturates, endogenous eicosanoids, and cardiovascular disease.

The role of dietary polyunsaturated fats in the prevention of human vascular disease has not been defined, but population and intervention studies have suggested that omega-3 fatty acids (FAs) from marine lipids may have a number of potentially beneficial effects. Eicosanoids are extremely potent autacoids made from polyunsaturated fatty acids and have effects on many vascular parameters, so that the physiological effects of dietary supplementation with polyunsaturated fats are often attributed to alterations in endogenous eicosanoid production. Few studies have attempted to correlate in vivo eicosanoid synthesis and functional effects during such dietary maneuvers, however. This article reviews the relationship between dietary polyunsaturates and endogenous eicosanoid synthesis in man, with particular emphasis on recent studies of the effects of omega-3 FAs. Data on omega-3 FAs and platelet-vascular interactions, blood pressure, and vascular reactivity in human subjects are also summarized, with interpretation of recent work addressing a number of controversial points. A discussion of the significance and future direction of such investigations concludes that further clinical trials in selected patient groups are warranted.

Polyunsaturated fatty acids and cardiovascular function in man.

Dietary supplementation with either n-6 or n-3 polyunsaturated fatty acids has been reported to lower blood pressure in man, and to confer benefits in atherosclerotic vascular disease. Such effects have been suggested to be due to altered in vivo production of vasoactive prostaglandins, but this hypothesis has not been tested previously. Epidemiologic data are often quoted as showing lower blood pressure in populations with a high consumption of fish, but such conclusions are less clear from the primary data. This review presents some of the population studies on the relationship between fish intake, hypertension and vascular disease, discusses problems in studying blood pressure in dietary intervention trials, and reviews the published literature on reducing blood pressure and vascular reactivity with dietary n-3 supplements in volunteers. Recent work addressing a number of controversial points is presented, with the conclusion that pharmacologic doses of n-3, but not n-6, fatty acids can lower blood pressure in humans, but probably do not do so through directly lowering vascular reactivity to catecholamines or increasing the synthesis of vasodilator prostaglandins.

Formation of PGI3 in the rat during dietary fish oil supplementation.

Conflicting results exist in the literature on the conversion of eicosapentaenoic acid (EPA) to trienoic prostaglandins and its influence on the formation of dienoic prostaglandins from arachidonic acid (AA). Tissues from animals fed fish oils produce little, if any, trienoic prostaglandins and reduced amounts of dienoic ones. Excretion of the major urinary metabolite of PGI2 is not reduced in humans taking fish oil, however, and substantial amounts of one derived from PGI3 have been found, by GC/MS. We have addressed this possible species difference by examining the urine of rats fed fish oil for 2.3 dinor-6-keto-PGF1 alpha and its delta 17 analog, formed from PGI2 and PGI3, respectively, and compared them with rats fed corn oil. Fatty acid differences in erythrocyte and aortic lipids were also determined. Rats fed fish oil do make PGI3 from eicosapentaenoic acid in vivo and do not suppress their production of PGI2, despite having more EPA than AA in aortic lipids.

The antihypertensive effects of fish oil. A controlled study of polyunsaturated fatty acid supplements in essential hypertension.

Both n-3 and n-6 polyunsaturated fats have been suggested to lower blood pressure, an effect ascribed to altered biosynthesis of eicosanoids. To test these hypotheses, we studied blood pressure and eicosanoid production during supplementation of dietary fat for four weeks in 32 men with mild essential hypertension. Supplementation was preceded and followed by four-week run-in and recovery periods. Groups of eight subjects received either 10 ml or 50 ml of fish oil (3 or 15 g of n-3 fatty acids) daily, 50 ml of safflower oil (39 g of n-6 fatty acids), or 50 ml of a mixture of oils that approximated the types of fat present in the American diet. The biosynthesis of eicosanoids was assessed by the measurement of urinary metabolites. Blood pressure decreased in the men who received the high dose of fish oil (systolic pressure by a mean of 6.5 mm Hg [P less than 0.03] and diastolic pressure by 4.4 mm Hg [P less than 0.015]), but not in the other groups. Although the formation of vasodilatory prostacyclins (prostaglandins I2 and I3) increased initially, this increase was not maintained as blood pressure fell. The level of thromboxane A2 metabolites fell; metabolites of thromboxane A3 were detected in the groups receiving fish oil. The formation of prostaglandin E2 increased during supplementation with safflower oil and tended to decrease with fish oil; no prostaglandin E3 metabolite was detected. Our data indicate that high doses of fish oil can reduce blood pressure in men with essential hypertension. However, the clinical usefulness and safety of fish oil in the treatment of hypertension will require further study.

Fish oils in cardiovascular disease.

Although several studies have reported an inverse relationship between the dietary history of fish intake and the prospective incidence of death from coronary heart disease, it is unclear whether these results represent the effects of n-3 fatty acids themselves or whether they merely reflect a more fundamental alteration in diet, such as a reduction in saturated fatty acids. n-3 fatty acids alter platelet eicosanoid formation, replacing arachidonate derived thromboxane (Tx)A2 with the biologically inert TxA3. However, they are a relatively inefficient approach to platelet inhibition. This is evident from results obtained in a model of coronary thrombosis followed by thrombolysis with tissue plasminogen activator (tPA). By contrast, high doses of n-3 fatty acids (15 g day-1) significantly reduced blood pressure in mild hypertensives in a double-blind, controlled study characterized by a prolonged run-in period and adequate follow-up. Fish oils inhibit the vascular proliferative response to injury in a variety of animal models, apparently independently of their effects on lipoprotein metabolism. Recent results imply that they may selectively interfere with the vascular expression of mitogenic peptides.

Bactericidal effects of polyunsaturated fatty acids.

Bactericidal effects of polyunsaturated fatty acids were investigated by using an in vitro killing assay. All gram-positive species tested were extremely susceptible to 10(-5) M arachidonic acid as were Neisseria, Branhamella, and Haemophilus spp. Pseudomonas aeruginosa and and members of the Enterobacteriaceae were resistant. The toxicity of polyunsaturated fatty acids for Staphylococcus aureus was dependent upon time, concentration, and fatty acid unsaturation. Arachidonic acid underwent peroxidation when incubated with S. aureus, but arachidonic acid peroxidation products had low bactericidal activity. Catalase protected S. aureus, whereas superoxide dismutase was ineffective. Scavengers of hydroxyl radicals or singlet oxygen or removal of halide ions had little effect on arachidonic acid-induced killing of bacteria, whereas transition metal chelators and some thiols were highly protective. S. aureus grown in iron-supplemented broth had increased iron content and arachidonic acid susceptibility. Ascorbate also potentiated arachidonic acid-induced killing of S. aureus. These observations indicate that bactericidal effects of polyunsaturated fatty acids are mediated by a peroxidative process involving H2O2 and bacterial iron.

In vivo indexes of platelet and vascular function during fish-oil administration in patients with atherosclerosis.

Populations that consume a diet rich in marine lipids may have a lower risk of atherosclerotic disease. Fish oil contains the N-3 polyunsaturated fatty acid eicosapentaenoate, and the biosynthesis of thromboxanes and prostacyclins from eicosapentaenoate (thromboxane A3 and prostaglandin I3), rather than from the usual precursor arachidonate (thromboxane A2 and prostaglandin I2), may help to reduce the risk. To examine this hypothesis, we studied the effect of eicosapentaenoate supplementation (10 g per day) for one month on the synthesis of thromboxanes and prostacyclins, as assessed by urinary metabolite excretion, in six patients with peripheral vascular disease and seven normal controls. Supplementation markedly increased the eicosapentaenoate content of phospholipids from red cells and platelets. Synthesis of the platelet agonist thromboxane A2, which was elevated in the patients at base line, declined by 58 percent during supplementation but did not reach normal values. The decline in thromboxane A2, which is synthesized from arachidonate, coincided with the formation of the inactive thromboxane A3, which is synthesized from eicosapentaenoate. A lower dose of eicosapentaenoate (1 g per day) was not sufficient to maintain the changes in thromboxane A2 synthesis. Platelet function was only moderately inhibited during eicosapentaenoate supplementation, consistent with incomplete suppression of thromboxane A2 synthesis. These studies show that a high dose of eicosapentaenoate alters the pattern of synthesis of thromboxanes and prostacyclins. However, effects comparable to those of aspirin require long-term administration in high doses. Whether other properties of fish oil might render it a more attractive antithrombotic therapy remains to be determined.