Effect of vasopressin on Na(+)-K(+)-2Cl(-) cotransporter (NKCC) and the signaling mechanisms on the murine late distal colon.

摘要: It has been demonstrated that the antidiuretic hormone vasopressin is able to regulate expression of Na-K-Cl cotransporters (NKCC1 and NKCC2) in kidney. The present study investigated effects long- short-term administration on NKCC possible signaling mechanism mouse distal colon using siRNA, real-time PCR, western blotting Ussing chambers method. results showed presence NKCC2 colon, which was verified with a siRNA technique. mRNA protein level significantly increased by about 40% 90% respectively response restricting water intake 1ml/day/20g for 7 days. In contrast, NKCC1 unchanged colon. To determine activation vitro, we found caused 3-fold increase phosphorylation, detected phosphospecific antibody R5. addition, chamber NKCC2, cAMP Ca(2+) pathway may be involved vasopressin-induced response. Further, adenylate cyclase inhibitor MDL-12330A PKA H89 chelator BAPTA-AM reversed induced phosphorylation 35%, 28% 42% suggesting stimulate mediated cAMP-PKA Collectively, these data suggest are stimulation, association enhanced activity vasopressin/cAMP pathways.