leukotriene-b4 and Ulcer

Small intestinal ulcers are frequent complications of therapy with nonsteroidal anti-inflammatory drugs (NSAIDs). We present here a genetic deficiency of eicosanoid biosynthesis that illuminates the mechanism of NSAID-induced ulcers of the small intestine.. Eicosanoids and metabolites were measured by isotope dilution with mass spectrometry. cDNA was obtained by reverse transcription and sequenced following amplification with RT-PCR.. We investigated the cause of chronic recurrent small intestinal ulcers, small bowel perforations, and gastrointestinal blood loss in a 45-year-old man who was not taking any cyclooxygenase inhibitor. Prostaglandin metabolites in urine were significantly depressed. Serum thromboxane B2 (TxB2) production was 4.6% of normal controls (P<0.006), and serum 12-HETE was 1.3% of controls (P<0.005). Optical platelet aggregation with simultaneous monitoring of ATP release demonstrated absent granule secretion in response to ADP and a blunted aggregation response to ADP and collagen, but normal response to arachidonic acid (AA). LTB4 biosynthesis by ionophore-activated leukocytes was only 3% of controls, and urinary LTE4 was undetectable. These findings suggested deficient AA release from membrane phospholipids by cytosolic phospholipase A2-alpha (cPLA2-alpha), which regulates cyclooxygenase- and lipoxygenase-mediated eicosanoid production by catalyzing the release of their substrate, AA. Sequencing of cPLA2-alpha cDNA demonstrated two heterozygous nonsynonymous single-base-pair mutations: Ser111Pro (S111P) and Arg485His (R485H), as well as a known single nucleotide polymorphism (SNP), Lys651Arg (K651R).. Characterization of this cPLA2-alpha deficiency provides support for the importance of prostaglandins in protecting small intestinal integrity and indicates that loss of prostaglandin biosynthesis is sufficient to produce small intestinal ulcers.

Topics: Arachidonic Acid; Base Pair Mismatch; Base Sequence; DNA, Complementary; Eicosanoids; Group IV Phospholipases A2; Humans; Intestinal Diseases; Intestine, Small; Leukotriene B4; Leukotriene E4; Male; Middle Aged; Polymorphism, Single Nucleotide; Ulcer

The effects of dietary pretreatment on longitudinal ulcers of the intestine induced by indomethacin given intracolonically were investigated in rats. The rats were pretreated with either standard diet or liquid meals. Intracolonic indomethacin (24 mg/kg/day) given for two days produced longitudinal ulcers and small scattered ulcers in the small intestine in the control rats that were receiving standard pelleted formula. Three days pretreatment with one of two types of liquid meals, low residual diet (LRD) or elemental diet (ED), significantly reduced the incidence (3% in ED group and 0% in the LRD group) and the length of the longitudinal ulcers in the small intestine. The caecum was affected in each dietary pretreatment group (67% in controls, 80% in LRD group, and 69% in ED group). Colonic ulcers that were located in a longitudinal fashion were found in 42% of LRD group, while these ulcers were less frequently found in the ED group (13%) and controls (0%). Development of ulcers in the caecum and in the colon of rats in ED and LRD groups was more delayed than that of small intestinal ulcers of control rats. In another experiment, pretreatment by ED significantly increased colonic tissue leukotriene B4 concentration when compared with that of controls. These findings suggest that the site of experimental enteropathy induced by indomethacin given intracolonically can be modified by dietary pretreatment. This animal model can be available for investigating differences in the pathophysiology of enteropathy according to the site of involvement.

Topics: Animals; Diet; Food, Formulated; Indomethacin; Intestinal Diseases; Intestinal Mucosa; Leukotriene B4; Leukotriene C4; Male; Rats; Rats, Wistar; Ulcer

The effect of highly purified eicosapentaenoic acid ethyl ester (EPA-E) on colitis was investigated using a guinea pig model. The technique for preparing a degraded carrageenan with a molecular weight of about 30,000 from commercial iota-carrageenan was first refined. When this degraded carrageenan was fed to guinea pigs, localized ulcerations occurred in the cecum with infiltration of numerous mononuclear phagocytes. Oral administration of 300 mg.kg-1.day-1 of EPA-E for 3 weeks significantly prevented the development of colitis. The amounts of prostaglandin E2, thromboxane B2, and leukotriene B4 released from the cecal mucosa were also measured. The release of prostaglandin E2 and thromboxane B2 was significantly decreased in the animals fed EPA-E compared with those given olive oil or a vehicle alone. In addition, there was a positive correlation between the amounts of these eicosanoids and the degree of ulcer formation. However, there was no difference in the amount of leukotriene B4 among various experimental groups of animals. Furthermore, EPA-E feeding induced a significant decrease in the level of arachidonic acid and a significant increase in that of EPA in peritoneal macrophages. These results suggest that EPA has a prophylactic effect on the development of carrageenan-induced colitis, which may be ascribed in part to reduced eicosanoid production.

Topics: Animals; Body Weight; Carrageenan; Cecal Diseases; Colitis; Dinoprostone; Dose-Response Relationship, Drug; Eicosapentaenoic Acid; Fatty Acids; Guinea Pigs; Leukotriene B4; Macrophages; Male; Occult Blood; Thromboxane B2; Ulcer

The ability of various leukotrienes, platelet-activating factor and N-formyl-methyl-leucyl-phenylalanine to augment colonic damage induced by 30% ethanol was investigated in the rat. Each of the mediators was tested at a dose of 2 nmol, administered intracolonically in the ethanol vehicle. When colonic damage was assessed 72 hr later, only leukotriene B4 significantly augmented damage compared to the controls. The incidence of ulcers increased from 35% in the control group to 90% in the group receiving leukotriene B4. Leukotriene B4 administration also resulted in significant increases in colonic myeloperoxidase activity and colonic leukotriene B4 synthesis. To assess the possible contribution of infiltrating neutrophils to the increase in colonic leukotriene B4 synthesis that accompanies colonic inflammation, colitis was induced in normal and neutropenic rats by intracolonic administration of trinitrobenzene sulfonic acid. Neutropenia was achieved by treatment with an antineutrophil serum. In the neutropenic animals killed 4 hr after induction of colitis significant changes in leukotriene B4 synthesis were not observed, whereas a fourfold increase was observed in the controls. From these studies we conclude the following: (1) leukotriene B4, at a dose of 2 nmol, can significantly potentiate the colonic ulceration induced by 30% ethanol; (2) this action of leukotriene B4 is not shared by the same dose of the other inflammatory mediators tested; and (3) infiltrating neutrophils are the major source of colonic leukotriene B4 synthesis in a rat model of colitis.

Topics: Administration, Rectal; Animals; Colitis; Drug Synergism; Ethanol; Leukotriene B4; Leukotrienes; Male; N-Formylmethionine Leucyl-Phenylalanine; Neutropenia; Platelet Activating Factor; Rats; Rats, Inbred Strains; Ulcer