Downregulation of epithelial sodium channel (ENaC) activity in human airway epithelia after low temperature incubation.
Downregulation of epithelial sodium channel (ENaC) activity in human airway epithelia after low temperature incubation.
复制标题
低温孵育后人气道上皮细胞中上皮钠通道（ENaC）活性的下调。
DOI：
10.1136/bmjresp-2020-000861
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发表时间：
2021-03
影响因子：
4.1
通讯作者：
Bratcher PE
中科院分区：
文献类型：
--
作者：
Yadav S;Shaughnessy CA;Zeitlin PL;Bratcher PE
在低温下孵育气道上皮细胞是囊性纤维化（CF）研究领域中常用的体外实验方法，以热稳定F508 del-CFTR并增加其功能表达。考虑到气道上皮细胞包括CFTR以外的许多离子转运蛋白，我们假设低温孵育对来自有和没有CF的个体的气道上皮细胞中CFTR独立的离子调节机制有影响。在空气-液体界面处分化后，将鼻上皮细胞在37°C或29°C（低温）下孵育48小时，然后在Ussing室中分析。虽然低温孵育后F508 del-CFTR活性增加，但非CF上皮中CFTR的活性不变。重要的是，在29°C下孵育的培养物在基线时表现出降低的跨上皮电位差（TEPD）和短路电流（Isc）。导致29°C培养物中基线TEPD和Isc降低的主要因素是上皮钠通道（ENaC）活性降低，这由对阿米洛利的反应性降低证明。在来源于非CF和CF供体的细胞中观察到这种效应。ENaC亚基β和γ的转录水平显著下调，这可能是ENaC活性降低的部分原因。我们推测，低温孵育可能是一个有用的实验范例，以减少ENaC活性在体外上皮细胞培养。
The incubation of airway epithelia cells at low temperatures is a common in vitro experimental approach used in the field of cystic fibrosis (CF) research to thermo-stabilise F508del-CFTR and increase its functional expression. Given that the airway epithelium includes numerous ion transporters other than CFTR, we hypothesised that there was an impact of low temperature incubation on CFTR-independent ionoregulatory mechanisms in airway epithelia derived from individuals with and without CF. After differentiation at the air–liquid interface, nasal epithelia were incubated at either 37°C or 29°C (low temperature) for 48 hours prior to analysis in an Ussing chamber. While F508del-CFTR activity was increased after low temperature incubation, activity of CFTR in non-CF epithelia was unchanged. Importantly, cultures incubated at 29°C demonstrated decreased transepithelial potential difference (TEPD) and short-circuit currents (Isc) at baseline. The predominant factor contributing to the reduced baseline TEPD and Isc in 29°C cultures was the reduced activity of the epithelial sodium channel (ENaC), evidenced by a reduced responsiveness to amiloride. This effect was observed in cells derived from both non-CF and CF donors. Significant transcriptional downregulation of ENaC subunits β and γ were observed, which may partially explain the decreased ENaC activity. We speculate that low temperature incubation may be a useful experimental paradigm to reduce ENaC activity in in vitro epithelial cultures.
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影响因子：
7.7
作者：
König, J;Schreiber, R;Kunzelmann, K
通讯作者：
Kunzelmann, K
DOI：
10.1073/pnas.111155398
发表时间：
2001-05-22
影响因子：
11.1
作者：
Askwith, CC;Benson, CJ;Snyder, PM
通讯作者：
Snyder, PM
DOI：
10.1152/ajpcell.1995.268.1.c243
发表时间：
1995-01-01
影响因子：
5.5
作者：
EGAN, ME;SCHWIEBERT, EM;GUGGINO, WB
通讯作者：
GUGGINO, WB
DOI：
10.1152/ajprenal.00195.2017
发表时间：
2018-03-01
影响因子：
4.2
作者：
Kashlan, Ossama B.;Kinlough, Carol L.;Kleyman, Thomas R.
通讯作者：
Kleyman, Thomas R.
DOI：
10.1152/ajpgi.1999.277.3.g709
发表时间：
1999-09-01
影响因子：
4.5
作者：
Mall, M;Bleich, M;Kunzelmann, K
通讯作者：
Kunzelmann, K