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首页> 外文期刊>Life sciences >Cocaine permeability and metabolism in colonic T-84 epithelial cell line.
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Cocaine permeability and metabolism in colonic T-84 epithelial cell line.

机译：可卡因在结肠T-84上皮细胞系中的渗透性和代谢。

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We studied the uptake, transport and metabolism of cocaine in human intestine using the colonic T-84 monolayers as a model. T-84 cells were grown in DMEM/Ham's F-12 medium containing 6% newborn calf serum at 37 degrees C on 1.0 microm collagen inserts. The cells develop into a polarized monolayer with the apical surface facing the upper chamber and the basolateral surface facing the lower. The monolayers develop a transepithelial resistance of > or = 600 ohms cm2 in 7 days. Varying concentrations of cocaine HCI was added in a serum free medium to the luminal side only, and after 30 min and 60 min samples from the luminal and serosal aspect were removed for analysis. Cocaine and its metabolites were measured by GC/MS. Cocaine transport across T-84 monolayers increased linearly with increasing concentration of cocaine, with no significant difference between 30 min and 60 min of exposure. Transepithelial resistance did not change even at 800 ng of cocaine suggesting no effect on monolayer viability. The metabolites, benzoylecgonine (BE) and ecgonine methyl ester (EME) but not norcocaine were detected in both luminal and serosal side. The concentrations of BE and EME in the luminal side were significantly higher than in the serosal. Combined recovery of cocaine, BE and EME from luminal and serosal sides were 52 - 80% of total added cocaine. While fresh medium did not metabolize cocaine, media previously exposed to the monolayer (cell-free medium) caused a significant breakdown into BE and EME, suggesting that esterases may be released into the medium. These results indicate transfer of cocaine across this model of intestinal epithelial cell line is by simple diffusion and is concentration dependent. These studies imply that cocaine in swallowed amniotic fluid can be absorbed by the fetal gastrointestinal tract.
机译：我们使用结肠T-84单层细胞作为模型研究了可卡因在人肠中的吸收，运输和代谢。 T-84细胞在含有6％新生小牛血清的DMEM / Ham's F-12培养基中于1.0微米胶原插入物上于37摄氏度生长。细胞发展成极化的单层，其顶表面面向上室，而基底外侧表面面向下室。单层在7天内产生>或= 600 ohms cm2的跨上皮电阻。仅在无血清培养基中向管腔侧添加不同浓度的可卡因HCl，并在30分钟和60分钟后从管腔和浆膜方面取出样品进行分析。可卡因及其代谢产物通过GC / MS测定。可卡因跨T-84单层的转运随着可卡因浓度的增加而线性增加，在30分钟和60分钟的暴露时间之间无显着差异。即使在800 ng可卡因下，跨上皮的耐药性也没有改变，表明对单层存活率没有影响。在管腔和浆膜侧均未检出代谢产物苯甲酰基芽子碱（BE）和芽子碱甲酯（EME），但未检测到降丁卡因。腔侧的BE和EME浓度显着高于浆膜侧。从腔和浆膜侧可卡因，BE和EME的总回收量为可卡因添加总量的52-80％。尽管新鲜培养基不会代谢可卡因，但先前暴露于单层的培养基（无细胞培养基）会导致BE和EME大量分解，这表明酯酶可能会释放到培养基中。这些结果表明可卡因在这种肠上皮细胞系模型中的转移是通过简单扩散并且是浓度依赖性的。这些研究表明，吞咽的羊水中的可卡因可以被胎儿胃肠道吸收。

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