epidermal-growth-factor and benzamil

Various stimuli, including hormones and growth factors, modulate epithelial sodium channels (ENaCs), which fine-tune Na(+) absorption in the kidney. Members of the EGF family are important for maintaining transepithelial Na(+) transport, but whether EGF influences ENaC, perhaps mediating salt-sensitive hypertension, is not well understood. Here, the ENaC inhibitor benzamil attenuated the development of hypertension in Dahl salt-sensitive rats. Feeding these salt-sensitive rats a high-salt diet led to lower levels of EGF in the kidney cortex and enhanced the expression and activity of ENaC compared with feeding a low-salt diet. To directly evaluate the role of EGF in the development of hypertension and its effect on ENaC activity, we infused EGF intravenously while continuously monitoring BP of the salt-sensitive rats. Infusion of EGF decreased ENaC activity, prevented the development of hypertension, and attenuated glomerular and renal tubular damage. Taken together, these findings indicate that cortical EGF levels decrease with a high-salt diet in salt-sensitive rats, promoting ENaC-mediated Na(+) reabsorption in the collecting duct and the development of hypertension.

Topics: Amiloride; Animals; Blood Pressure; Disease Models, Animal; Epidermal Growth Factor; Epithelial Sodium Channels; Hypertension; Kidney; Rats; Rats, Inbred Dahl; Sodium Chloride, Dietary

The objective of this study was to investigate acute and long-term effects of epidermal growth factor (EGF) and transforming growth factor alpha (TGFalpha) on basal ion transport activity of glandular endometrial epithelial cells in primary culture. The effects of EGF on insulin-dependent regulation of Na+ transport across this epithelium was also investigated. Addition of 1.6 nM EGF or 2 nM TGFalpha to the basolateral, but not the apical, solution inhibited both basal and insulin-stimulated Na+ transport with a maximum response within 45-60 min. This effect was mimicked by the calcium ionophore ionomycin. Incubation with EGF for 4 days inhibited insulin-stimulated Na absorption in a concentration-dependent fashion with an IC(50) value of 0.3 nM. Experiments using amphotericin B-permeabilized monolayers demonstrated that EGF inhibited Na transport by decreasing apical membrane Na conductance without affecting insulin-dependent stimulation of the Na+-K+ ATPase. Addition of EGF or TGFalpha for 24 h resulted in increased basal Cl- secretion in addition to inhibition of Na absorption. The EGF-induced increase in Cl- secretion was inhibited in part by indomethacin, suggesting that long-term regulation by EGF involves stimulation of arachidonic acid synthesis and prostaglandin release. The EGF-induced increase in indomethacin-insensitive Cl- secretion was prevented by the protein synthesis inhibitor cyclohexamide, and by the DNA transcription inhibitor actinomycin D indicating that EGF-stimulated anion secretion required DNA transcription and protein synthesis. The results of these studies demonstrated that the basal transport properties of endometrial epithelial cells are differentially regulated by EGF, TGFalpha, and insulin.

Topics: Amiloride; Animals; Biological Transport; Cell Membrane; Cell Membrane Permeability; Cells, Cultured; Chlorides; Cyclooxygenase Inhibitors; Dactinomycin; Endometrium; Epidermal Growth Factor; Epithelial Cells; Female; Indomethacin; Insulin; Ionomycin; Kinetics; Nitrobenzoates; Prostaglandins; Sodium; Sodium-Potassium-Exchanging ATPase; Swine; Transforming Growth Factor alpha

The effect of Lys-bradykinin on net Na+ influx in serum-deprived cultured human fibroblasts (HSWP cells) was measured. It was found that Lys-bradykinin stimulates net Na+ influx with a K1/2 of 1 nM. When Lys-bradykinin was combined with epidermal growth factor, vasopressin and insulin, the net Na+ influx stimulated was comparable with that measured in response to 10% serum. The above combination of growth factors also was found to stimulate DNA synthesis to 92% of the serum-stimulated levels in HSWP cells and to support cell growth over a period of 6 days. In addition, it was observed that the Na+ influx stimulated by Lys-bradykinin or by the combination of four growth factors was completely inhibited by the amiloride analog benzamil. Thus Lys-bradykinin presumably stimulates the same Na+ transport system as is stimulated by serum. Finally, the present results suggest that an increase in Na+ influx always accompanies DNA synthesis in HSWP cells. On the basis of these observations, it can be hypothesized that Na+ influx is a necessary event to stimulate DNA synthesis.

Topics: Amiloride; Cell Line; DNA Replication; Epidermal Growth Factor; Fibroblasts; Humans; Insulin; Kallidin; Sodium; Vasopressins