The effect of transient receptor potential vanilloid 4 on the intestinal epithelial barrier and human colonic cells was affected by tyrosine-phosphorylated claudin-7
The effect of transient receptor potential vanilloid 4 on the intestinal epithelial barrier and human colonic cells was affected by tyrosine-phosphorylated claudin-7
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DOI：
10.1016/j.biopha.2019.109697
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发表时间：
2020-02-01
影响因子：
7.5
通讯作者：
Xia, Xian-Ming
中科院分区：
文献类型：
--
作者：
Huang, Yuan-Yuan;Li, Jing;Xia, Xian-Ming
TRPV4是一种非选择性阳离子通道，在胃肠道中激活TRPV4可引起小鼠实验性结肠炎。先前的一项研究发现，酪氨酸磷酸化的cladin -7在实验性结肠炎中增加。酪氨酸磷酸化的claudin-7与TRPV4之间的关系尚不清楚。在本研究中，我们通过用谷氨酸取代酪氨酸在210位置开发了一个cld7- 7突变体，命名为cld7-Y210E结肠细胞。我们发现GSK1016790A激活TRPV4增加了对照结肠细胞单层的通透性，而TRPV4拮抗剂HC067047则降低了通透性。在cld7-Y210E结肠细胞单层中，GSK1016790A和HC067047处理的通透性没有差异。GSK1016790A在对照结肠细胞中增加了cludin -7在细胞膜上的聚集，而HC067047则减弱了这种作用。在cld7-Y210E结肠细胞中，GSK1016790A和HC067047均未明显改变cld7的聚集。GSK1016790A和HC067047均未改变载体结肠细胞中的TRPV4蛋白水平。在cld7-野生结肠细胞中，GSK1016790A不改变TRPV4蛋白水平，而HC067047增加了TRPV4蛋白水平。cld7-Y210E结肠细胞中TRPV4蛋白水平升高，GSK1016790A降低，HC067047进一步降低。在GSK1016790A处理的对照结肠细胞中，钙内流没有明显改变。然而，GSK1016790A显著增加了cld7-Y210E结肠细胞的钙内流。我们得出结论，酪氨酸磷酸化的cludin -7影响TRPV4调节的肠上皮屏障、TRPV4介导的钙内流以及TRPV4在人结肠细胞中的蛋白表达。我们认为酪氨酸磷酸化的cladin -7影响TRPV4调节的肠上皮屏障，这可能与TRPV4表达和TRPV4介导的钙内流有关。
TRPV4 is a type of nonselective cation channel, and activation of TRPV4 in the gastrointestinal tract causes experimental colitis in mice. A previous study found that tyrosine-phosphorylated claudin-7 is increased in experimental colitis. The relationship between tyrosine-phosphorylated claudin-7 and TRPV4 remains undefined. In the present study, we developed a claudin-7 mutant by replacing tyrosine with glutamic acid at position 210, named cld7-Y210E colonic cells. We found that activation of TRPV4 by GSK1016790A increased the permeability of control colonic cell monolayers, which was decreased by the TRPV4 antagonist HC067047. In monolayers of cld7-Y210E colonic cells, no differences in permeability were found between GSK1016790A and HC067047 treatments. GSK1016790A increased the aggregation of claudin-7 at the cell membrane in control colonic cells, and the effect was diminished by HC067047. In cld7-Y210E colonic cells, neither GSK1016790A nor HC067047 apparently changed the aggregation of claudin-7. Neither GSK1016790A nor HC067047 altered the TRPV4 protein level in vector colonic cells. In cld7-wild colonic cells, GSK1016790A did not alter the TRPV4 protein level, while HC067047 increased the TRPV4 protein level. The TRPV4 protein level was increased in cld7-Y210E colonic cells, decreased by GSK1016790A and further decreased by HC067047. Calcium influx was not significantly changed in the control colonic cells treated with GSK1016790A. However, GSK1016790A significantly increased calcium influx in cld7-Y210E colonic cells. We concluded that tyrosine-phosphorylated claudin-7 affects the TRPV4-modulated intestinal epithelial barrier, TRPV4-mediated calcium influx, and the protein expression of TRPV4 in human colonic cells. We suggest that tyrosine-phosphorylated claudin-7 affects the TRPV4-modulated intestinal epithelial barrier, which might be related to TRPV4 expression and TRPV4-mediated calcium influx.