Verotoxin induces hemorrhagic lesions in rat small intestine. Temporal alteration of vasoactive substances.
E. coli O157:H7 produces a cytotoxin active against Vero cells that has been termed verotoxin. In this study, we demonstrated that local intraarterial injection of verotoxin induced a decrease in blood flow and an increase in hemorrhagic lesions in rat small intestine. Significant increases in the area of hemorrhagic lesions were observed at 120 min after verotoxin injection. These lesions were produced by either verotoxin 1 or 2, but verotoxin 2 produced more extensive lesions. The temporal alteration of vasoactive substances in microcirculatory beds was determined after the administration of culture filtrate of E. coli O157:H7. Tissue-type plasminogen activator activity in regional plasma was significantly elevated as early as 30 min, suggesting that local fibrinolytic activation mediated by microvascular endothelium occurred. There was also early elevation of platelet-activating factor content in the ileal mucosa and its level remained significantly elevated thereafter. Intestinal blood flow, as determined by a laser Doppler flowmeter, started to decrease at about 45 min. The platelet-activating factor antagonist CV6209 was shown to attenuate the decrease in blood flow as well as the development of hemorrhagic lesions, demonstrating that platelet-activating factor is an important mediator for the microcirculatory damage. Accumulation of neutrophils demonstrated by myeloperoxidase activity in the intestinal mucosa and overproduction of oxygen-radicals from neutrophils of the mesenteric veins determined by the luminol-dependent chemiluminescence assay were observed at 60 min, corresponding with the decreased blood flow. Platelet-activating factor may be closely involved in the process of leukocyte accumulation and increased oxygen radical generation, because CV6209 also significantly attenuated these changes.(ABSTRACT TRUNCATED AT 250 WORDS)
Tashiro
H
H
Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
Miura
S
S
Kurose
I
I
Fukumura
D
D
Suzuki
H
H
Suematsu
M
M
Yoshioka
M
M
Tsuchiya
M
M
Kai
A
A
Kudoh
Y
Y
D000818:Animals; D001427:Bacterial Toxins / Q000008:administration & dosage / Q000633:toxicity*; D004768:Enterotoxins / Q000008:administration & dosage / Q000633:toxicity*; D004926:Escherichia coli; D005609:Free Radicals / Q000378:metabolism; D006471:Gastrointestinal Hemorrhage / Q000139:chemically induced*; D007269:Injections, Intra-Arterial; D007421:Intestine, Small / Q000098:blood supply*; D017078:Laser-Doppler Flowmetry; D008297:Male; D008833:Microcirculation / Q000187:drug effects; D009504:Neutrophils / Q000378:metabolism; D009195:Peroxidase / Q000032:analysis; D010972:Platelet Activating Factor / Q000032:analysis* / Q000037:antagonists & inhibitors; D011726:Pyridinium Compounds / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D022622:Shiga Toxin 1; D010959:Tissue Plasminogen Activator / Q000097:blood*
D001427:Bacterial Toxins; D004768:Enterotoxins; D005609:Free Radicals; D010972:Platelet Activating Factor; D011726:Pyridinium Compounds; D022622:Shiga Toxin 1; C053321:CV 6209; D009195:Peroxidase; D010959:Tissue Plasminogen Activator