Tomita Mikio, Ohkubo Rie, Hayashi Masahiro
Department of Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Japan.
Drug Metab Pharmacokinet. 2004 Feb;19(1):33-40. doi: 10.2133/dmpk.19.33.
To clarify whether lipopolysaccharide (LPS) is transported in rat intestinal epithelial cells, the transport of FITC-LPS across colonic epithelial cells in normal and LPS-exposured rats using a diffusion chamber was examined. The expression of CD14 and Toll-like receptor 4 (TLR4) was also examined. Rats were given 10 mg/kg LPS i.p. injection at 4 hr prior to the isolation of colonic epithelial tissues. The permeation rate across colonic mucosa by FITC-LPS was several times greater in the mucosal to serosal (M to S) direction than in the opposite direction in both normal and LPS-exposured rats. Increased M to S permeation by FITC-LPS was evident at 37 degrees C, but not at 4 degrees C. The permeability of FITC-LPS in both the M to S and S to M directions was inhibited by unlabeled LPS, anti-CD14 antibody or anti-TRL4 antibody in normal rat. In LPS-exposured rat, the inhibition in the M to S direction was observed by anti-TLR4 antibody, but not by unlabeled LPS and anti- CD14 antibody. In contrast, the permeability in the S to M direction was decreased only by unlabeled LPS in LPS-exposured rat. In normal rat, the expression of CD14 and TLR4 was found in the mucosal and serosal sides. In LPS-exposured rat, the expression of CD14 was not observed in the mucosal side. The electrophysiological parameters by LPS exposure remain unchanged. These findings suggest the possibility that colonic epithelial cells contain specific transport systems for LPS, one of which shows some degree of substrate specificity with the interaction of CD14 and/or that of TLR4.
为了阐明脂多糖(LPS)是否在大鼠肠道上皮细胞中运输,使用扩散室检测了正常大鼠和暴露于LPS的大鼠中FITC-LPS跨结肠上皮细胞的运输情况。还检测了CD14和Toll样受体4(TLR4)的表达。在分离结肠上皮组织前4小时,给大鼠腹腔注射10 mg/kg LPS。在正常大鼠和暴露于LPS的大鼠中,FITC-LPS跨结肠黏膜的渗透速率在黏膜到浆膜(M到S)方向比在相反方向大几倍。FITC-LPS在37℃时M到S的渗透增加明显,但在4℃时不明显。在正常大鼠中,未标记的LPS、抗CD14抗体或抗TRL4抗体可抑制FITC-LPS在M到S和S到M方向的通透性。在暴露于LPS的大鼠中,抗TLR4抗体可观察到M到S方向的抑制作用,但未标记的LPS和抗CD14抗体则无此作用。相反,在暴露于LPS的大鼠中,仅未标记的LPS可降低S到M方向的通透性。在正常大鼠中,在黏膜和浆膜侧均发现了CD14和TLR4的表达。在暴露于LPS的大鼠中,在黏膜侧未观察到CD14的表达。LPS暴露后的电生理参数保持不变。这些发现提示结肠上皮细胞可能含有LPS的特异性转运系统,其中之一在与CD14和/或TLR4相互作用时表现出一定程度的底物特异性。