Diclofenac toxicity in human intestine ex vivo is not related to the formation of intestinal metabolites
Diclofenac toxicity in human intestine ex vivo is not related to the formation of intestinal metabolites
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DOI：
10.1007/s00204-014-1242-6
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发表时间：
2015-01-01
影响因子：
6.1
通讯作者：
Groothuis, Geny M. M.
中科院分区：
文献类型：
--
作者：
Niu, Xiaoyu;de Graaf, Inge A. M.;Groothuis, Geny M. M.
使用双氯芬酸（DCF），一种非甾体抗炎药，与胃肠道副作用的高发率有关。啮齿类动物的体内研究表明，肝脏或肠道产生的DCF反应性代谢物可能是造成这种毒性的原因。本研究以18例人类供体空肠制备的精确切肠片（PCIS）作为离体模型，研究DCF肠道代谢物是否与人体肠道毒性有关。DCF （a parts per thousand yen400 a mu M）浓度范围内培养PCIS至24 h。DCF （a parts per thousand yen400 a mu M）对肠道产生直接毒性，表现为ATP消耗、形态损伤、caspase 3激活和乳酸脱氢酶渗漏。HPLC法检测了PCIS产生的3种主要代谢物（4′-羟基DCF、5-羟基DCF和DCF酰基葡萄糖醛酸酯）。免疫组化染色检测蛋白质加合物，发现其与肠道代谢物相关。DCF对每个样品的毒性相似，而不考虑它们之间代谢的差异。与100 A μ M DCF相比，400 A μ M DCF培养的切片产生的代谢物较少。代谢抑制剂如酮康唑、西咪替丁或冰片的加入减少了代谢物的形成，但增加了毒性。结果表明，DCF可以在治疗相关浓度下直接诱导人PCIS的肠道毒性，而不依赖于肝脏产生或肠内形成的反应性代谢物4'-OH DCF、5-OH DCF或双氯芬酸酰基葡萄糖醛酸。
The use of diclofenac (DCF), a nonsteroidal anti-inflammatory drug, is associated with a high prevalence of gastrointestinal side effects. In vivo studies in rodents suggested that reactive metabolites of DCF produced by the liver or the intestine might be responsible for this toxicity. In the present study, precision-cut intestinal slices (PCIS) prepared from the jejunum of 18 human donors were used as an ex vivo model to investigate whether DCF intestinal metabolites are responsible for its intestinal toxicity in man. PCIS were incubated with a concentration range of DCF (0-600 A mu M) up to 24 h. DCF (a parts per thousand yen400 A mu M) caused direct toxicity to the intestine as demonstrated by ATP depletion, morphological damage, caspase 3 activation, and lactate dehydrogenase leakage. Three main metabolites produced by PCIS (4'-hydroxy DCF, 5-hydroxy DCF, and DCF acyl glucuronide) were detected by HPLC. Protein adducts were detected by immunohistochemical staining and showed correlation with the intestinal metabolites. DCF induced similar toxicity to each of the samples regardless of the variation in metabolism among them. Less metabolites were produced by slices incubated with 400 A mu M DCF than with 100 A mu M DCF. The addition of the metabolic inhibitors such as ketoconazole, cimetidine, or borneol decreased the metabolite formation but increased the toxicity. The results suggest that DCF can induce intestinal toxicity in human PCIS directly at therapeutically relevant concentrations, independent of the reactive metabolites 4'-OH DCF, 5-OH DCF, or diclofenac acylglucuronide produced by the liver or formed in the intestine.