We used patch-clamp and western blot to test whether PGF2α stimulates the basolateral 10-pS Cl channel and thiazide-sensitive Na-Cl cotransporter (NCC) in distal convoluted tubule (DCT) via FP receptor (FP-R). The single-channel and whole cell recordings demonstrated that PGF2α stimulated the 10-pS Cl channel in DCT. The stimulatory effect of PGF2α on the Cl channel was mimicked by FP-R agonist, Latanoprost, but was abrogated by blocking FP-R with AL8810. Also, the effect of PGF2α on the Cl channel in the DCT was recapitulated by stimulating protein kinaseC (PKC) but was blocked by inhibiting PKC. Furthermore, the inhibition of p38-mitogen-activated protein kinase (MAPK) but not ERK blocked the effect of PGF2α on 10-pS Cl. Inhibition of NADPH oxidase (NOX) also abrogated the stimulatory effect of PGF2α on the 10-pS Cl channel while adding 10μM H2O2 mimicked the stimulatory effect of PGF2α on the 10-pS Cl channel. Moreover, superoxide-related-species may mediate the stimulatory effect of PGF2α on the 10-pS Cl channel because the stimulatory effect of PGF2α and H2O2 was not additive. Western blot showed that infusion of PGF2α in vivo not only increased the expression of FP-R, but also increased the expression of total NCC (tNCC) and phosphorylated NCC (pNCC). We conclude that PGF2α stimulates the basolateral 10-pS Cl channel in the DCT by activating FP-R through PKC/p38 MAPK and NOX-dependent pathways. The stimulatory effects of PGF2α on the Cl channel and NCC may contribute to PGF2α induced increases in NaCl reabsorption in the DCT.

中文翻译：

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PGF2α刺激远曲小管中的10-pS Cl-通道和噻嗪类敏感的Na + -Cl-共转运蛋白。

我们使用膜片钳和蛋白质印迹来测试PGF2α是否通过FP受体（FP-R）刺激远端回旋小管（DCT）中的基底外侧10-pS Cl通道和噻嗪类敏感的Na-Cl共转运蛋白（NCC）。单通道和全细胞记录表明，PGF2α刺激了DCT中的10-pS Cl通道。FP-R激动剂Latanoprost模仿了PGF2α对Cl通道的刺激作用，但被AL8810阻断FP-R消除了PGF2α对Cl通道的刺激作用。同样，通过刺激蛋白激酶C（PKC）概括了PGF2α对DCT中Cl通道的作用，但通过抑制PKC阻止了其生长。此外，对p38促分裂原活化蛋白激酶（MAPK）的抑制而非对ERK的抑制也阻止了PGF2α对10-pS Cl的作用。NADPH氧化酶（NOX）的抑制作用也消除了PGF2α对10-pS Cl通道的刺激作用，而添加10μMH2O2模仿了PGF2α对10-pS Cl通道的刺激作用。此外，与超氧化物有关的物种可能介导PGF2α对10-pS Cl通道的刺激作用，因为PGF2α和H2O2的刺激作用不是累加的。免疫印迹表明，体内输注PGF2α不仅增加了FP-R的表达，而且还增加了总NCC（tNCC）和磷酸化NCC（pNCC）的表达。我们得出结论，PGF2α通过通过PKC / p38 MAPK和NOX依赖性途径激活FP-R刺激DCT的基底外侧10-pS Cl通道。PGF2α对Cl通道和NCC的刺激作用可能有助于DCT中PGF2α诱导的NaCl重吸收增加。