Involvement of Rho-kinase and tyrosine kinase in hypotonic stress-induced ATP release in bovine aortic endothelial cells
Involvement of Rho-kinase and tyrosine kinase in hypotonic stress-induced ATP release in bovine aortic endothelial cells
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DOI：
10.1111/j.1469-7793.2001.0759e.x
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发表时间：
2001-05-01
影响因子：
5.5
通讯作者：
Ito, Y
中科院分区：
文献类型：
--
作者：
Koyama, T;Oike, M;Ito, Y
1.低渗应激诱导牛主动脉内皮细胞（BAECs）释放ATP，随后出现Ca 2+振荡。研究了低渗应激诱导ATP释放的细胞机制.低渗应激诱导至少两种蛋白质的酪氨酸磷酸化，110和150 kDa。酪氨酸激酶抑制剂除莠霉素A和tyrphostin 46抑制酪氨酸激酶可抑制低渗应激引起的ATP释放和ATP介导的Ca 2+振荡. ATP的释放也被肉毒杆菌毒素C3预处理的细胞抑制，并增加溶血磷脂酸。此外，Rho激酶选择性抑制剂Y-27632预处理也能抑制低渗应激诱导的ATP释放和Ca 2+振荡，提示Rho激酶的激活可能参与了低渗应激诱导的ATP释放.低渗应激还诱导了肌动蛋白细胞骨架的瞬时重排，酪氨酸激酶抑制剂Y-27632和细胞松弛素B抑制了这种重排。然而，细胞松弛素B预处理既不抑制低渗应激诱导的ATP释放，也不抑制Ca 2+振荡.这些结果表明，酪氨酸激酶和Rho-Rho-激酶途径参与低渗应激诱导的ATP释放和肌动蛋白重排，但肌动蛋白聚合是不需要的BAECs中的ATP释放。
1. Hypotonic stress induces ATP release followed by Ca2+ oscillations in bovine aortic endothelial cells (BAECs). We have investigated the cellular mechanism of the hypotonic stress-induced ATP release.2. Hypotonic stress induced tyrosine phosphorylation of at least two proteins, of 110 and 150 kDa. Inhibition of tyrosine kinase by the tyrosine kinase inhibitors herbimycin A and tyrphostin 46 prevented ATP release and ATP-mediated Ca2+ oscillations induced by hypotonic stress.3. ATP release was also inhibited by the pretreatment of the cells with botulinum toxin C3, and augmented by lysophosphatidic acid. Furthermore, pre-treating the cells with Y-27632, a selective inhibitor of Rho-kinase, also suppressed the hypotonic stress-induced ATP release and Ca2+ oscillations, indicating that Rho-mediated activation of Rho-kinase may be involved in the hypotonic: ATP release.4. Hypotonic stress also induced a transient rearrangement of the actin cytoskeleton, which was suppressed by the tyrosine kinase inhibitors Y-27632 and cytochalasin B. However, pretreatment of the cell with cytochalasin B inhibited neither the hypotonic stress-induced ATP release nor the Ca2+ oscillations.5. These results indicate that tyrosine kinase and the Rho-Rho-kinase pathways are involved in hypotonic stress-induced ATP release and actin rearrangement, but actin polymerization is not required for ATP release in BAECs.