Water extracts of Helicobacter pylori delay healing of chronic gastric ulcers in rats: role of cytokines and gastrin-somatostatin link.
BACKGROUND
Helicobacter pylori (Hp) is considered as a major risk factor of peptic ulcer, but the pathogenic mechanism of its action has not been fully explained.


OBJECTIVE
This study was designed: (1) to compare the ulcer healing effects of water extract (WE) obtained from type-I cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) expressing Hp and from type-II CagA- and VacA-negative Hp strain with those of vehicle (saline), and (2) to determine the alterations in gastric secretion, gastric blood flow (GBF) and expression of Hp-related cytokines during the ulcer healing in rats treated with toxigenic (type-I) and non-toxigenic (type-II) Hp-derived WE.


METHODS
Gastric ulcers were produced by serosal application of acetic acid in rats with or without gastric fistula treated with vehicle (saline) or WE originating from type-I or type-II Hp administered intragastrically on days 1, 3, 5 and 7 upon ulcer induction. On days 3, 9 and 15, animals were lightly anesthetized with ether, the abdomen was opened and the GBF was measured by the H2-gas clearance technique in the ulcer area and non-ulcerated mucosa. Venous blood was withdrawn for the measurement of plasma cytokine (IL-1beta and TNFalpha) levels and plasma and gastric contents were also collected for gastrin and somatostatin determination by specific radioimmunoassay.


RESULTS
Gastric ulcers healed gradually in vehicle-treated controls and the ulcer area on days 3, 9 and 15 was reduced by 12, 43 and 92%, respectively. In rats treated with WE of type-I Hp, ulcer healing was significantly delayed, and gastritis and infiltration of ulcerated gastric mucosa with inflammatory cells were observed histologically. The prolongation of ulcer healing by WE of both Hp strains was accompanied by a marked fall in the GBF at the ulcer margin and transient hyposecretion especially in rats given WE of type-I Hp strain. On day 15 of ulcer healing, the plasma concentration of IL-1beta and TNFalpha was negligible in vehicle control rats, but it was significantly elevated particularly in rats treated with WE of type-I Hp. RT-PCR analysis revealed that mucosal expression of IL-1beta and TNFalpha mRNA was significantly upregulated in the gastric mucosa of rats treated with either toxigenic or non-toxigenic Hp WE. The plasma gastrin level was significantly higher and the luminal concentration of somatostatin was significantly lower in rats treated with Hp-WE than in vehicle-treated controls and these alterations were more pronounced in rats treated with WE type-I than type-II Hp.


CONCLUSIONS
WE of toxigenic Hp strain delays ulcer healing due to the reduction in the gastric microcirculation at the ulcer margin, the overexpression of inflammatory cytokines and the impairment of the gastrin-somatostatin link.
Brzozowski
T
T
Department of Physiology and Pathomorphology, Jagiellonian University School of Medicine, Cracow, Poland.
Konturek
P C
PC
Konturek
S J
SJ
Kwiecien
S
S
Pajdo
R
R
Karczewska
E
E
Stachura
J
J
Hahn
E G
EG
D000818:Animals; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000633:toxicity; D005753:Gastric Mucosa / Q000098:blood supply / Q000378:metabolism; D005755:Gastrins / Q000096:biosynthesis / Q000502:physiology*; D016481:Helicobacter Infections / Q000503:physiopathology*; D016480:Helicobacter pylori / Q000472:pathogenicity*; D007375:Interleukin-1 / Q000096:biosynthesis / Q000502:physiology*; D008297:Male; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013004:Somatostatin / Q000096:biosynthesis / Q000502:physiology*; D013276:Stomach Ulcer / Q000382:microbiology / Q000503:physiopathology*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000502:physiology*; D014945:Wound Healing
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D005755:Gastrins; D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C074689:VacA protein, Helicobacter pylori; C080866:cagA protein, Helicobacter pylori; D013004:Somatostatin