The Journal of Physiological Sciences

, Volume 64, Issue 6, pp 433–443 | Cite as

Insulin is involved in transcriptional regulation of NKCC and the CFTR Cl channel through PI3K activation and ERK inactivation in renal epithelial cells

Original Paper


It is is well known that insulin stimulates glucose transport and epithelial Na+ channel (ENaC)-mediated Na+ reabsorption; however, the action of insulin on Cl secretion is not fully understood. In this study, we investigated the action of insulin on Na+–K+–2Cl cotransporter (NKCC)-mediated Cl secretion in epithelial A6 cells. Interestingly, insulin treatment remarkably enhanced the forskolin-stimulated Cl secretion associated with an increase in apical Cl conductance by upregulating mRNA expression of both CFTR and NKCC, although insulin treatment alone had no effect on the basal Cl secretion or apical Cl conductance without forskolin application. We next elucidated a role of phosphoinositide 3-kinase (PI3K) in the insulin-induced enhancement of the Cl secretion, since insulin actually activated PI3K, resulting in activation of Akt, a downstream molecule of PI3K. LY294002 (a PI3K inhibitor) reduced the Cl secretion by suppressing mRNA expression of NKCC, whereas insulin still had a stimulatory action on mRNA expression of CFTR even in the presence of LY294002. On the other hand, we found that a MEK inhibitor (PD98059) further enhanced the insulin-stimulated CFTR mRNA expression and the Cl secretion in forskolin-stimulated A6 cells and that insulin induced slight, transient activation of ERK followed by significant inactivation of ERK. These observations suggest that: (1) insulin respectively upregulates mRNA expression of NKCC and CFTR through activation of PI3K and inactivation of ERK; (2) insulin signals on mRNA expression of NKCC and CFTR are not enough to stimulate transepithelial Cl secretion, but enhance the stimulatory action of cAMP on transepithelial Cl secretion.


Insulin NKCC CFTR Cl secretion 



This work was supported by Grants-in-Aid from the Japan Society of the Promotion of Science (24590283 to NN, 25670111 to YM), Salt Science (1235 to YM and NN) and Cell Research Conference to YM.


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Copyright information

© The Physiological Society of Japan and Springer Japan 2014

Authors and Affiliations

  1. 1.Department of Molecular Cell Physiology, Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyotoJapan
  2. 2.Department of Bio-Ionomics, Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyotoJapan
  3. 3.Japan Institute for Food Education and HealthSt. Agnes’ UniversityKyotoJapan
  4. 4.Saisei Mirai ClinicsMoriguchiJapan

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