Swelling-Induced, Cftr-Independent Atp Release from a Human Epithelial Cell Line
Swelling-Induced, Cftr-Independent Atp Release from a Human Epithelial Cell Line
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人上皮细胞系肿胀诱导的、不依赖 Cftr 的 Atp 释放
DOI：
--
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Y. Okada
中科院分区：
文献类型：
--
作者：
A. Hazama;Takahiro Shimizu;Y. Ando‐Akatsuka;S. Hayashi;Shoko Tanaka;E. Maeno;Y. Okada
为了研究肿胀诱导的三磷酸腺苷释放途径与体积敏感的氯−通道之间的可能关系，我们测量了在渗透膨胀时释放的三磷酸腺苷的细胞外浓度和缺乏囊性纤维化跨膜电导调节因子表达的人上皮细胞株--肠上皮细胞系的全细胞体积敏感的氯-−电流。通过荧光素/荧光素酶分析，在低渗刺激(56-80%渗透压)后的几分钟内观察到ATP的显著释放。羧酸盐类似物氯-−通道阻断剂5-硝基-2-(3-苯丙氨基)-苯甲酸酯以浓度依赖的方式抑制三磷酸腺苷的释放，半数抑制浓度为6.3μM。然而，二苯乙烯类氯−通道阻断剂4-乙酰氨基-4‘-异硫氰基二苯乙烯(100μM)和花生四烯酸(100μM)对肿胀诱导的三磷酸腺苷释放无影响，格列本脲(500μM)和花生四烯酸(100μM)也不能有效地阻断体积敏感的外向整流性氯通道。Gd3+是一种牵张激活的通道阻断剂，它以浓度依赖的方式抑制肿胀引起的−释放，而三价稀土不能抑制VSORClATP电流。在渗透肿胀的情况下，利用表达在∼12细胞中的P2X2受体的生物传感器技术，发现细胞表面附近的局部三磷酸腺苷浓度达到了μ13 Mp。我们已经提出了抑制肿胀诱导的肠道407细胞释放ATP的抗体。早期用抗体处理几乎完全抑制肿胀诱导的−释放，而VSORClATP通道的活性不受抗体预处理的影响。综合以上结果，我们得出以下结论：第一，在缺乏cftr的人上皮细胞系中，渗透肿胀诱导细胞释放三磷酸腺苷，并使细胞表面的三磷酸腺苷浓度增加到10μM以上，足以刺激嘌呤能受体；第二，三磷酸腺苷的释放途径不同于容量敏感的外向整流氯−通道的孔道；第三，三磷酸腺苷的释放不是激活氯离子−通道的先决条件。
To examine a possible relation between the swelling-induced ATP release pathway and the volume-sensitive Cl− channel, we measured the extracellular concentration of ATP released upon osmotic swelling and whole-cell volume-sensitive Cl− currents in a human epithelial cell line, Intestine 407, which lacks expression of cystic fibrosis transmembrane conductance regulator (CFTR). Significant release of ATP was observed within several minutes after a hypotonic challenge (56–80% osmolality) by the luciferin/luciferase assay. A carboxylate analogue Cl− channel blocker, 5-nitro-2-(3-phenylpropylamino)-benzoate, suppressed ATP release in a concentration-dependent manner with a half-maximal inhibition concentration of 6.3 μM. However, swelling-induced ATP release was not affected by a stilbene-derivative Cl− channel blocker, 4-acetamido-4′-isothiocyanostilbene at 100 μM. Glibenclamide (500 μM) and arachidonic acid (100 μM), which are known to block volume-sensitive outwardly rectifying (VSOR) Cl− channels, were also ineffective in inhibiting the swelling-induced ATP release. Gd3+, a putative blocker of stretch-activated channels, inhibited swelling-induced ATP release in a concentration-dependent manner, whereas the trivalent lanthanide failed to inhibit VSOR Cl− currents. Upon osmotic swelling, the local ATP concentration in the immediate vicinity of the cell surface was found to reach ∼13 μM by a biosensor technique using P2X2 receptors expressed in PC12 cells. We have raised antibodies that inhibit swelling-induced ATP release from Intestine 407 cells. Earlier treatment with the antibodies almost completely suppressed swelling-induced ATP release, whereas the activity of VSOR Cl− channel was not affected by pretreatment with the antibodies. Taking the above results together, the following conclusions were reached: first, in a CFTR-lacking human epithelial cell line, osmotic swelling induces ATP release and increases the cell surface ATP concentration over 10 μM, which is high enough to stimulate purinergic receptors; second, the pathway of ATP release is distinct from the pore of the volume-sensitive outwardly rectifying Cl− channel; and third, the ATP release is not a prerequisite to activation of the Cl− channel.
cAMP对分泌Cl-的上皮细胞和3T3成纤维细胞胞内和胞外ATP含量的影响。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Takahashi,T;Matsushita,K;Welsh,MJ;Stokes,JB
通讯作者：
Stokes,JB
DOI：
10.1152/ajpheart.1998.275.5.h1726
发表时间：
1998-11-01
影响因子：
4.8
作者：
Sprague, RS;Ellsworth, ML;Lonigro, AJ
通讯作者：
Lonigro, AJ
DOI：
10.1073/pnas.90.1.312
发表时间：
1993-01-01
影响因子：
11.1
作者：
ABRAHAM, EH;PRAT, AG;CANTIELLO, HF
通讯作者：
CANTIELLO, HF