The effect of extracellular pH changes on intracellular pH and nitric oxide concentration in endothelial and smooth muscle cells from rat aorta

It has been known for more than a century that pH changes can alter vascular tone. However, there is no consensus about the effects of pH changes on vascular response. In this study, we investigated the effects of extracellular pH (pHo) changes on intracellular pH (pHi) and intracellular nitric oxid...

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Bibliographic Details
Published in:PloS one 2013-05, Vol.8 (5), p.e62887-e62887
Main Authors: Capellini, Verena K, Restini, Carolina B A, Bendhack, Lusiane M, Evora, Paulo R B, Celotto, Andréa C
Format: Article
Language:English
Subjects:
Acetic acid
Acid-base status
Acidification
Analysis of Variance
Anatomy & physiology
Animals
Aorta
Aorta - cytology
Benzopyrans - metabolism
Biology
Confocal microscopy
Coronary vessels
Cross sections
Cytometry
Endothelial cells
Endothelial Cells - chemistry
Endothelial Cells - metabolism
Endothelium
Flow Cytometry
Fluorescence
Fluorescent dyes
Fluorescent indicators
Hydrogen ions
Hydrogen-Ion Concentration
Intracellular
Laboratories
Medicine
Metabolism
Microscopy
Microscopy, Confocal
Muscles
Myocytes, Smooth Muscle - chemistry
Myocytes, Smooth Muscle - metabolism
Naphthols - metabolism
Nitric oxide
Nitric Oxide - metabolism
Ostomy
pH effects
Pharmaceutical sciences
Physiology
Potassium
Rats
Rhodamines - metabolism
Rodents
Smooth muscle
Surgery
Veins & arteries
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Summary:It has been known for more than a century that pH changes can alter vascular tone. However, there is no consensus about the effects of pH changes on vascular response. In this study, we investigated the effects of extracellular pH (pHo) changes on intracellular pH (pHi) and intracellular nitric oxide concentration ([NO]i) in freshly isolated endothelial cells and cross sections from rat aorta. The HCl was used to reduce the pHo from 7.4 to 7.0 and from 7.4 to 6.5; the NaOH was used to increase the pHo from 7.4 to 8.0 and from 7.4 to 8.5. The fluorescent dyes 5-(and-6)-carboxy SNARF-1, acetoxymethyl ester, acetate (SNARF-1) and diaminofluorescein-FM diacetate (DAF-FM DA) were employed to measure the pHi and [NO]i, respectively. The fluorescence intensity was measured in freshly isolated endothelial cells by flow cytometry and in freshly obtained aorta cross sections by confocal microscopy. The endothelial and vascular smooth muscle pHi was increased at pHo 8.5. The extracellular acidification did not change the endothelial pHi, but the smooth muscle pHi was reduced at pHo 7.0. At pHo 8.5 and pHo 6.5, the endothelial [NO]i was increased. Both extracellular alkalinization and acidification increased the vascular smooth muscle [NO]i. Not all changes in pHo did result in pHi changes, but disruption of acid-base balance in both directions induced NO synthesis in the endothelium and/or vascular smooth muscle.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0062887