PHYSIOLOGICAL AND CYTOTOXIC EFFECTS OF CA-2+ IONOPHORES ON CACO-2 PARACELLULAR PERMEABILITY - RELATIONSHIP OF CA-2+(45) EFFLUX TO CR-51 RELEASE

被引:14
作者
RUTTEN, MJ
COGBURN, JN
SCHASTEEN, CS
SOLOMON, T
机构
[1] UNIV KANSAS,MED CTR,DEPT PHYSIOL,KANSAS CITY,KS 66103
[2] UNIV KANSAS,MED CTR,DEPT MED,KANSAS CITY,KS 66103
[3] MONSANTO CO,MONSANTO CORP RES LABS,ST LOUIS,MO 63166
关键词
IONOMYCIN; A23187; CACO-2; PARACELLULAR PERMEABILITY; CA-2+(45) EFFLUX; CR-51; RELEASE; LACTATE DEHYDROGENASE; CYTOTOXIC;
D O I
10.1159/000138793
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
The human intestinal cell line, Caco-2, and the Ca2+ ionophores, A23187 and ionomycin, were used to determine the interrelationships of Ca-45(2+) efflux, transepithelial electrical resistance (R(t)), and [H-3]-mannitol flux to changes in Cr-51 release and lactate dehydrogenase (LDH) activity. Treatment of Caco-2 monolayers with ionomycin at concentrations of between 0.25 and 2.50-mu-mol/l showed similar Ca-45(2+) efflux rate constants and coefficients. Analysis of the control and ionomycin-induced Ca-45(2+) efflux values showed the data to best fit a three Ca2+ compartmental model. All changes in Caco-2 R(t) and [H-3]-mannitol flux were reversible with no significant increases in Cr-51 release with ionomycin concentrations of less-than-or-equal-to 2.5-mu-mol/l. Caco-2 monolayers treated with ionomycin at concentrations of between 5.0 and 50.0-mu-mol/l showed rapid non-exponential Ca-45(2+) effluxes with irreversible changes in R(t), [H-3]-mannitol flux, and significant increases in Cr-51 release. There was no change in media LDH activity using either ionomycin or A23187 at concentrations of up to 50-mu-mol/l for 60 min. The results of our study show that: (1) disruption of Ca2+ homeostasis in Caco-2 cells will occur with the addition of Ca2+ ionophores at concentrations of > 2.50-mu-mol/l; (2) high concentrations (> 2.5-mu-mol/l) of ionomycin will cause non-exponential Ca-45(2+) efflux rates with irreversible changes in transcellular R(t) and C-14-mannitol flux, and (3) early signs of Ca2+ ionophore-induced damage can be detected by Cr-51 release from Caco-2 cells into the media and not by changes in LDH media activity.
引用
收藏
页码:156 / 168
页数:13
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