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N-脱乙酰酶/磺基转移酶和3-O-磺基转移酶在大鼠II型肺泡细胞中的表达。

Expression of N-deacetylase/sulfotransferase and 3-O-sulfotransferase in rat alveolar type II cells.

作者信息

Li Z Y, Hirayoshi K, Suzuki Y

机构信息

Department of Ultrastructural Research, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto 606, Japan.

出版信息

Am J Physiol Lung Cell Mol Physiol. 2000 Aug;279(2):L292-301. doi: 10.1152/ajplung.2000.279.2.L292.

Abstract

Basal laminae beneath alveolar type I cells are suggested to contain highly sulfated heparan sulfate-containing proteoglycans (PGs), and cultured type II cells accumulate highly sulfated matrices. To characterize the regulation of PG synthesis during the transition from type II cells to type I cells, we examined mRNA expression of N-deacetylase/sulfotransferase (NST) and 3-O-sulfotransferase (3-OST), two enzymes specific for heparan sulfate synthesis. We found that both freshly isolated and cultured type II cells expressed NST and 3-OST as shown by in situ hybridization. Expression of surfactant-associated protein A, B, and C mRNAs, determined by semiquantitative PCR, decreased during culture. Expression of type I cell marker T1alpha mRNA increased except in cells cultured on an Engelbrecht-Holm-Swarm gel. Expression of NST was dependent on cell density and matrix and was intense in conditions where cells spread fully, whereas 3-OST expression was unchanged in the conditions examined. The PG sulfation inhibitor sodium chlorate significantly inhibited cultured type II cell spreading, and this inhibition was reversed by sodium sulfate. These results suggest that highly sulfated PGs modified by NST are necessary for the spreading of cells during transdifferentiation of type II cells to mature type I cells.

摘要

肺泡I型细胞下方的基底层被认为含有高度硫酸化的含硫酸乙酰肝素蛋白聚糖(PGs),并且培养的II型细胞会积累高度硫酸化的基质。为了表征从II型细胞向I型细胞转变过程中PG合成的调控,我们检测了N-脱乙酰酶/磺基转移酶(NST)和3-O-磺基转移酶(3-OST)的mRNA表达,这两种酶是硫酸乙酰肝素合成所特有的。我们发现,通过原位杂交显示,新鲜分离的和培养的II型细胞均表达NST和3-OST。通过半定量PCR测定,表面活性剂相关蛋白A、B和C的mRNA表达在培养过程中下降。I型细胞标志物T1α mRNA的表达增加,但在Engelbrecht-Holm-Swarm凝胶上培养的细胞除外。NST的表达取决于细胞密度和基质,在细胞充分铺展的条件下表达强烈,而3-OST的表达在所检测的条件下没有变化。PG硫酸化抑制剂氯酸钠显著抑制培养的II型细胞铺展,而这种抑制作用可被硫酸钠逆转。这些结果表明,由NST修饰的高度硫酸化的PGs对于II型细胞向成熟I型细胞转分化过程中的细胞铺展是必需的。

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