Bile acid-induced depolarization of mitochondrial membrane potential preceding cell injury in cultured gastric mucosal cells.
Changes in energy metabolism elicited by sodium taurocholate and their relation to cell viability were determined in gastric mucosal cells. Cultured mucosal cells were labelled with rhodamine-123, a mitochondrial energization-sensitive fluorescence probe, or by propidium iodide, a fluorochrome which labels the nuclei of non-viable cells. The cells were observed under a fluorescence microscope with a laser scanning confocal imaging system. After the addition of sodium taurocholate at concentrations > 5 mol/L, mucosal cells showed a rapid and significant decrease in rhodamine-123 fluorescence. A decrease to 40% of the pretreated values at 30 min was seen with a concentration of sodium taurocholate of 7.5mmol/L. A marked increase in the percentage of propidium iodide-positive cells was noted when the concentration of sodium taurocholate exceeded 5mmol/L. However, the extent of the decrease in rhodamine-123 fluorescence was always greater than the increase in the percentage of propidium iodide-positive cells, suggesting that most of these gastric mucosal cells remained viable. It is therefore suggested that the decrease in rhodamine-123 fluorescence is largely due to the disturbed oxidative phosphorylation of mitochondria. Pretreatment of gastric mucosal cells with low concentrations of ethanol resulted in a significant cytoprotective effect against sodium taurocholate injury with significant prevention of a decrease in rhodamine-123 fluorescence. It is concluded that sodium taurocholate induces a depolarization of the mitochondrial membrane potential preceding cell injury and that the cytoprotective effect of ethanol relates to its attenuation of the uncoupling effect.
Miura
S
S
Department of Internal Medicine, School of Medicine, Keio University, Japan.
Fukumura
D
D
Shiozaki
H
H
Suzuki
M
M
Kurose
I
I
Suematsu
M
M
Tsuchiya
M
M
Ishii
H
H
D000818:Animals; D000963:Antimetabolites; D002452:Cell Count; D002470:Cell Survival; D002478:Cells, Cultured; D002756:Cholagogues and Choleretics / Q000494:pharmacology*; D005753:Gastric Mucosa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D011817:Rabbits; D020112:Rhodamine 123; D012235:Rhodamines / Q000494:pharmacology; D013656:Taurocholic Acid / Q000494:pharmacology*
D000963:Antimetabolites; D002756:Cholagogues and Choleretics; D012235:Rhodamines; D020112:Rhodamine 123; D013656:Taurocholic Acid