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在可渗透支持物上培养的T84细胞形成的单层的电学参数和水渗透特性。

Electrical parameters and water permeability properties of monolayers formed by T84 cells cultured on permeable supports.

作者信息

Ozu M, Toriano R, Capurro C, Parisi M

机构信息

Laboratorio de Biomembranas, Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina.

出版信息

Braz J Med Biol Res. 2005 Jan;38(1):133-40. doi: 10.1590/s0100-879x2005000100020. Epub 2005 Jan 18.

Abstract

T84 is an established cell line expressing an enterocyte phenotype whose permeability properties have been widely explored. Osmotic permeability (POSM), hydraulic permeability (PHYDR) and transport-associated net water fluxes (JW-transp), as well as short-circuit current (ISC), transepithelial resistance (RT), and potential difference (deltaVT) were measured in T84 monolayers with the following results: POSM 1.3 +/- 0.1 cm.s-1 x 10-3; PHYDR 0.27 +/- 0.02 cm.s-1; RT 2426 +/- 109 omega.cm2, and deltaVT 1.31 +/- 0.38 mV. The effect of 50 microM 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one (DCEBIO), a "net Cl- secretory agent", on T84 cells was also studied. We confirm the reported important increase in ISC induced by DCEBIO which was associated here with a modest secretory deltaJW-transp. The present results were compared with those reported using the same experimental approach applied to established cell lines originating from intestinal and renal epithelial cells (Caco-2, LLC-PK1 and RCCD-1). No clear association between PHYDR and RT could be demonstrated and high PHYDR values were observed in an electrically tight epithelium, supporting the view that a "water leaky" barrier is not necessarily an "electrically leaky" one. Furthermore, the modest secretory deltaJW-transp was not consistent with previous results obtained with RCCD-1 cells stimulated with vasopressin (absorptive fluxes) or with T84 cells secreting water under the action of Escherichia coli heat stable enterotoxin. We conclude that, while the presence of aquaporins is necessary to dissipate an external osmotic gradient, coupling between water and ion transport cannot be explained by a simple and common underlying mechanism.

摘要

T84是一种已确立的表达肠上皮细胞表型的细胞系,其通透性特性已得到广泛研究。在T84单层细胞中测量了渗透通透性(POSM)、水力通透性(PHYDR)和与转运相关的净水通量(JW-transp),以及短路电流(ISC)、跨上皮电阻(RT)和电位差(deltaVT),结果如下:POSM为1.3±0.1 cm·s-1×10-3;PHYDR为0.27±0.02 cm·s-1;RT为2426±109 Ω·cm2,deltaVT为1.31±0.38 mV。还研究了50 microM 5,6-二氯-1-乙基-1,3-二氢-2H-苯并咪唑-2-酮(DCEBIO),一种“净Cl-分泌剂”,对T84细胞的影响。我们证实了报道的DCEBIO诱导的ISC显著增加,在此与适度的分泌性deltaJW-transp相关。将本研究结果与使用相同实验方法应用于源自肠道和肾上皮细胞的已确立细胞系(Caco-2、LLC-PK1和RCCD-1)所报道的结果进行了比较。未发现PHYDR与RT之间存在明确关联,并且在电紧密上皮中观察到高PHYDR值,支持了“水渗漏”屏障不一定是“电渗漏”屏障的观点。此外,适度的分泌性deltaJW-transp与先前用血管加压素刺激的RCCD-1细胞(吸收通量)或在大肠杆菌热稳定肠毒素作用下分泌水的T84细胞所获得的结果不一致。我们得出结论,虽然水通道蛋白的存在对于消除外部渗透梯度是必要的,但水和离子转运之间的偶联不能用简单且共同的潜在机制来解释。

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