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18日龄胎鼠远端肺组织培养上皮单层的生物电特性

Bioelectric properties of cultured epithelial monolayers from distal lung of 18-day fetal rat.

作者信息

Barker P M, Stiles A D, Boucher R C, Gatzy J T

机构信息

Department of Medicine, University of North Carolina, Chapel Hill 27599-7020.

出版信息

Am J Physiol. 1992 May;262(5 Pt 1):L628-36. doi: 10.1152/ajplung.1992.262.5.L628.

Abstract

In vivo studies of fetal sheep suggest that the liquid present in the lumen of the lung throughout fetal life is derived from Cl- secretion by the pulmonary epithelium. Monolayer preparations of enriched epithelial cells from distal fetal rat (18-day gestation) lung, grown in serum-free media, were histologically similar to acinar (prealveolar) structures of fresh tissue. In Ussing chambers, basal transepithelial potential difference (PD), calculated equivalent short-circuit current (Ieq), and transepithelial resistance (R) were 4.4 +/- 0.3 mV (matrix positive), 35.6 +/- 1.6 microA/cm2, and 120.0 +/- 4.0 omega cm2, respectively. Ouabain (10(-3) M) eliminated 57% of basal Ieq within 30 min, amiloride (10(-4) M) induced a 13% fall in Ieq, and phlorizin (10(-4) M) had no effect on bioelectric properties. Diphenylamine-2-carboxylate (DPC, 3 x 10(-3) M) inhibited Ieq by 50%. Bumetanide had no effect on baseline bioelectric parameters. The hyperpolarization that accompanied apical or bilateral replacement of Cl- and was enhanced by terbutaline suggested an apical Cl- permselectivity. Effects of Na+ replacement on amiloride-pretreated monolayers were consistent with Na(+)-dependent Cl- secretion or amiloride-insensitive pathways. Under these growth conditions, this preparation exhibits bioelectric characteristics that are compatible with Cl- secretion and Na+ absorption. The mechanism of Cl- secretion may be similar to that of airways but is uniquely bumetanide insensitive.

摘要

对胎羊的体内研究表明,在整个胎儿期肺腔中存在的液体源自肺上皮细胞的氯离子分泌。在无血清培养基中培养的来自妊娠18天的胎鼠肺远端的富集上皮细胞单层制剂,在组织学上与新鲜组织的腺泡(肺泡前)结构相似。在尤斯灌流小室中,基础跨上皮电位差(PD)、计算得出的等效短路电流(Ieq)和跨上皮电阻(R)分别为4.4±0.3 mV(基质为正)、35.6±1.6 μA/cm²和120.0±4.0 Ω·cm²。哇巴因(10⁻³ M)在30分钟内使基础Ieq消除57%,氨氯吡脒(10⁻⁴ M)使Ieq下降13%,而根皮苷(10⁻⁴ M)对生物电特性无影响。二苯胺-2-羧酸盐(DPC,3×10⁻³ M)使Ieq抑制50%。布美他尼对基线生物电参数无影响。顶端或双侧置换氯离子时伴随的超极化并被特布他林增强,提示顶端氯离子的选择通透性。钠离子置换对经氨氯吡脒预处理的单层制剂的影响与钠离子依赖性氯离子分泌或对氨氯吡脒不敏感的途径一致。在这些生长条件下,该制剂表现出与氯离子分泌和钠离子吸收相符的生物电特性。氯离子分泌的机制可能与气道相似,但独特之处在于对布美他尼不敏感。

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