Effects of genotypically different strains of Helicobacter pylori on human microvascular endothelial cells in vitro.
Helicobacter pylori induces a number of disturbances in rodent gastric microcirculation in vivo. These events may result from direct necrotic or apoptotic damage to endothelial cells. This study therefore aimed to investigate the effects of genotypically different H. pylori strains on microvascular endothelial cell (MVEC) viability in vitro. Four H. pylori extracts were prepared from strains with different cagA or vacA status. MVECs were plated into 96-well plates and coincubated with 50 microl of extract or vehicle for 24, 48, 72, or 96 hr. An MPP assay quantified overall MVEC viability. The dual labeling of MVECs with propidium iodide and Hoechst 33342 distinguished between necrotic and apoptotic cell death, respectively, and allowed total number of viable cells to be determined. All strains of H. pylori decreased cell viability after 72 and 96 hr. Neither necrosis or apoptosis was observed. Counting total number of viable cells revealed decreased cell proliferation with all strains when compared to controls, again reaching significance at 72 and 96 hr. In conclusion, both the MTT assay and the diret cell counting technique demonstrated that all H. pylori strains induced cytostatic but not cytotoxic effects on MVECs. This suggests that microcirculatory disturbances observed in vivo may not be the result of direct endothelial cell damage. However, inhibition of angiogenesis may explain why ulcer healing is delayed in H. pylori-infected patients.
Department of Surgical and Anaesthetic Sciences, Royal Hallamshire Hospital, University of Sheffield, UK.
N
N
Kalia
C
C
Jones
D K
DK
Bardhan
M W
MW
Reed
J C
JC
Atherton
N J
NJ
Brown
D017209:Apoptosis; D001562:Benzimidazoles; D002470:Cell Survival; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000382:microbiology / Q000473:pathology*; D005838:Genotype; D016480:Helicobacter pylori / Q000235:genetics*; D006801:Humans; D008833:Microcirculation / Q000473:pathology; D009336:Necrosis; D011419:Propidium; D013270:Stomach / Q000098:blood supply*
D001562:Benzimidazoles; D011419:Propidium; C017807:bisbenzimide ethoxide trihydrochloride