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证据表明Ⅰ型肺泡上皮细胞存在钠-葡萄糖协同转运蛋白。

Evidence for Na+-glucose cotransporter in type I alveolar epithelium.

机构信息

Dipartimento di Fisiologia Umana, Università degli Studi di Milano, 20133 Milan, Italy.

出版信息

Histochem Cell Biol. 2010 Aug;134(2):129-36. doi: 10.1007/s00418-010-0725-7. Epub 2010 Jul 13.

Abstract

Functional evidence of Na(+)-glucose cotransport in rat lung has been provided by Basset et al. (J. Physiol. 384:325-345, 1987). By autoradiography [(3)H]phloridzin binding has been found confined to alveolar epithelial type II cells in mouse and rabbit lungs (Boyd, J. Physiol. 422: 44P, 1990). In this research we checked by immunofluorescence whether Na(+)-glucose cotransporter (SGLT1) is also expressed in alveolar type I cells. Lungs of anesthetized rats and lambs were fixed by paraformaldehyde, perfused in pulmonary artery, or instilled into a bronchus, respectively. Tissue blocks embedded in paraffin or frozen were sectioned. Two specific anti-SGLT1 antibodies for rat recognizing aminoacid sequence 402-420, and 546-596 were used in both species. Bound primary antibody was detected by secondary antibody conjugated to fluorescein isothiocianate or Texas red, respectively. In some sections cellular nuclei were also stained. In rats alveolar type I cells were identified by fluorescent Erythrina cristagalli lectin. Sections were examined by confocal laser-scanning microscope. Both in rats and lambs alveolar epithelium was stained by either antibody; no labeling occurred in negative controls. Hence, SGLT1 appears to be also expressed in alveolar type I cells. This is functionally relevant because type I cells provide 95-97% of alveolar surface, and SGLT1, besides contributing to removal of lung liquid under some circumstances, keeps low glucose concentration in lining liquid, which is useful to prevent lung infection.

摘要

巴塞特等人提供了大鼠肺中钠-葡萄糖协同转运的功能证据。[3H]根皮苷结合的放射自显影研究发现,小鼠和兔肺中的 [(3)H]根皮苷结合仅限于肺泡上皮 II 型细胞(博伊德,J. Physiol. 422: 44P,1990)。在这项研究中,我们通过免疫荧光检查 Na(+)-葡萄糖协同转运蛋白(SGLT1)是否也在肺泡 I 型细胞中表达。用多聚甲醛固定麻醉大鼠和羔羊的肺,分别通过肺动脉灌注或支气管内灌注进行固定。将石蜡或冷冻包埋的组织块切成切片。两种针对大鼠 SGLT1 的特异性抗体,一种识别氨基酸序列 402-420,另一种识别 546-596,在两种物种中均有使用。通过与异硫氰酸荧光素或 Texas Red 偶联的二级抗体检测结合的初级抗体。在一些切片中还对细胞核进行了染色。在大鼠中,肺泡 I 型细胞通过荧光 Erythrina cristagalli 凝集素进行鉴定。用共聚焦激光扫描显微镜检查切片。在大鼠和羔羊中,肺泡上皮均被任一抗体染色;阴性对照无标记。因此,SGLT1 似乎也在肺泡 I 型细胞中表达。这在功能上是相关的,因为 I 型细胞提供了 95-97%的肺泡表面,而 SGLT1 除了在某些情况下有助于清除肺液外,还能保持衬液中的低葡萄糖浓度,这有助于防止肺部感染。

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