PGF2α stimulates 10-pS Cl channel and thiazide-sensitive Na-Cl cotransporter (NCC) in distal convoluted tubule.
PGF2α stimulates 10-pS Cl channel and thiazide-sensitive Na-Cl cotransporter (NCC) in distal convoluted tubule.
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DOI：
10.1152/ajprenal.00287.2020
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发表时间：
2020-07
期刊：
影响因子：
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通讯作者：
Li-jun Wang;Yu Xiao;Jing Fang;Jun-Lin Wang;Hao Zhang;Xinxin Meng;R. Gong;R. Gu
中科院分区：
文献类型：
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作者：
Li-jun Wang;Yu Xiao;Jing Fang;Jun-Lin Wang;Hao Zhang;Xinxin Meng;R. Gong;R. Gu
我们用膜片钳和蛋白质印迹法检测前列腺素F_2α是否通过FP受体(FP-R)兴奋远端曲管的10-PS氯通道和硫氮化物敏感的钠-氯协同转运体。单通道和全细胞记录表明，PGF_2α刺激DCT的10-pS-氯通道。前列环素α对氯离子通道的兴奋作用可被FP-R激动剂拉坦前列素模拟，但可被AL8810阻断。PgF_2α对DCT中氯通道的作用可通过刺激蛋白激酶C来实现，但可被抑制蛋白激酶C所阻断。此外，抑制p38丝裂原活化蛋白激酶而不是ERK可阻断PGF_2α对10-PS_1的作用。抑制NADPH氧化酶(NOx)也可减弱PgF_2α对10-pS氯离子通道的刺激作用，而加入10μM H_2O_2可模拟PgF_2α对10-pS氯离子通道的刺激作用。此外，由于PgF_2α和H_2O_2的刺激作用不是相加的，因此PgF_2α和H_2O_2的刺激作用不是相加的，因此，超氧阴离子相关物质可能介导了PgF_2对10-pS的氯离子通道的刺激作用。Western印迹结果显示，体内注射前列腺素F2α不仅能增加FP-R的表达，还能增加总NCC(TnCC)和磷酸化NCC(PNCC)的表达。结论：前列腺素F2MAPK/p38MAPK和α依赖的通路激活FP-R，从而刺激双侧丘脑基底外侧10-PSCl通道。PgF_2α对氯通道和NCC的刺激作用可能是PgF_2α引起DCT重吸收增加的原因之一。
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.