PHARMACOLOGICAL PROFILE OF INHIBITION OF 2',7'-BIS(2-CARBOXYETHYL)-5(6)-CARBOXYFLUORESCEIN EFFLUX IN HUMAN HCT-8 INTESTINAL EPITHELIAL-CELLS
PHARMACOLOGICAL PROFILE OF INHIBITION OF 2',7'-BIS(2-CARBOXYETHYL)-5(6)-CARBOXYFLUORESCEIN EFFLUX IN HUMAN HCT-8 INTESTINAL EPITHELIAL-CELLS
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DOI：
10.1016/0006-2952(91)90389-m
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发表时间：
1991-12-11
影响因子：
5.8
通讯作者：
HIRST, BH
中科院分区：
文献类型：
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作者：
COLLINGTON, GK;ALLEN, CN;HIRST, BH
2',7'-二（2-羧基乙基）-5(6)-羧基荧光素（BCECF）从人HCT-8肠上皮细胞外排具有时间依赖性，5小时后76%的荧光色素在细胞外。抑制这种外排的药理学特征已经被研究，重点是调节阴离子运输的药物。BCECF外排对0.5 mM吲哚美辛的抑制敏感（20 μ m时抑制50%），使外排减少到叠氮和2-脱氧-d -葡萄糖耗尽ATP后观察到的值。前列腺素不能逆转吲哚美辛对BCECF外排的抑制作用。二苯乙烯衍生物4-乙酰氨基-4'-异硫氰基-2-2'-二硫苯基二苯乙烯和4,4'-二异硫氰基-2,2'-二硫苯基二苯乙烯即使在1mm时也只能部分抑制BCECF流出。probenecid和5-硝基-2-（3-苯基丙基-氨基）苯甲酸酯仅在1 mM时降低BCECF外排。阳离子剂vinblastine作为BCECF外排抑制剂的活性与吲哚美辛一样（10 μ m抑制50%），放线菌素D也是一个很好的抑制剂（100 μ m抑制50%）。其他几种阳离子药物，包括硝苯地平、阿米洛利和利血平，在浓度高达1mm时，作为BCECF外排抑制剂无效。因此，抑制BCECF外排的药理学特征并不完全等同于任何公认的运输系统。细胞松弛素B和氯喹等药物不影响BCECF的外排，这表明内体的积聚和随后的排出不是分泌途径。BCECF可能是上皮细胞分泌解毒系统的底物。
The efflux of 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) from human HCT-8 intestinal epithelial cultured cells was time-dependent, and after 5 hr 76% of the fluorochrome was extracellular. The pharmacological profile for inhibition of this efflux has been investigated, focusing on agents which modulate anion transport. BCECF efflux was sensitive to inhibition by 0.5 mM indomethacin (50% inhibition at 20-mu-M) which reduced efflux to values observed after depletion of ATP with azide and 2-deoxy-D-glucose. Indomethacin inhibition of BCECF efflux was not reversed with prostaglandin. The stilbene derivatives 4-acetamido-4'-isothiocyano-2-2'-disulphonic stilbene and 4,4'-diisothiocyano-2,2'-disulphonic stilbene only resulted in partial inhibition of BCECF efflux, even at 1 mM. Furosemide, bumetamide, probenecid and 5-nitro-2-(3-phenylpropyl-amino)-benzoate only reduced BCECF efflux at 1 mM. The cationic agent vinblastine was as active as indomethacin as an inhibitor of BCECF efflux (50% inhibition with 10-mu-M) while actinomycin D was also a good inhibitor (50% inhibition with 100-mu-M). Several other cationic agents, including nifedipine, amiloride and reserpine, were ineffective as inhibitors of BCECF efflux in concentrations up to 1 mM. Thus, the pharmacological profile for inhibition of BCECF efflux does not fully equate with any recognised transport system. Agents such as cytochalasin B and chloroquine did not effect BCECF efflux suggesting accumulation and subsequent discharge from endosomes is not a pathway for secretion. BCECF may be a substrate for a cellular secretory detoxifying system in epithelial cells.