mouse jejunum. Sections of mouse jejunum were mounted in Ussing chambers and matched for transepithelial electrical resistance 1 set was taken care of with 1 nM AII to the serosal side and mucosal to serosal and s to m fluxes measured over the subsequent 30 min. Values are signifies SEM for six separate experiments. p 0. 05 pared with untreated tissues by paired Students T test. All of these agents except PD98059 inhibited the AII stim ulation of NHE3 activity soon after 1 hour results that had been paralleled by their results on AII stimulated api cal surface NHE3 To find out if the long-term adjustments in NHE3 expres sion had been also mediated by form I receptor stimulation, cells were pretreated with losartan or PD123319 and stim ulated with AII for 24 hours and Na fluxes measured. Inhibition with the form I receptor blocked the AII stimu lated Na flux boost even though PD123319 had no result six. 59 0. 68, losartan AII four.
29 0. 54, and PD123319 AII 6. 36 0. 79 Consequently the long-term effects of AII on Caco2BBE NHE3 can also be mediated by type I receptor stimulation. Discussion The role of AII in regulation of blood stress is properly established, nevertheless, its actions are likely to take place via several mechanisms such as results on vascu lar smooth muscle and endothelium selleck chemicals GSK2118436 AII can also influence salt and water homeostasis by its actions on renal Na reabsorption Furthermore, AII stimulates aldosterone production from the adrenal gland which is a major regulator of renal and intestinal Na transport The current scientific studies demonstrate that AII has direct effects on intestinal epithelial Na transport that are consistent with its wanted result to increase fluid absorption. Angiotensin II increases NHE3 gene transcription Angiotensin II increases NHE3 gene transcription.
Monolayers had been taken care of with AII for various times and RNA harvested and analyzed for NHE3 mRNA by actual time PCR. GAPDH was applied as being a constitutive mRNA manage and NHE3 mRNA selleck chemical increases calculated by the Ct strategy Values are implies SEM for 4 sepa charge experiments. P 0. 05 P 0. 01 P 0. 001 pared with zero time untreated management by examination of variance implementing a Bonferroni correction. Monolayers were transfected with plasmids containing a 2200 bp segment on the rat NHE3 gene promoter linked to firefly luciferase cDNA and a further together with the thymidine kinase promoter linked to Renilla luciferase as a constitutive handle. Cells were taken care of with AII 24 hours soon after transfection and monolayers have been harvested and luciferase activities measured following 24 hours. Values are suggests SEM for 4 separate experiments. p 0. 05 pared with untreated zero time handle by evaluation of variance working with a Bonferroni correction.