Abstract
To create in vitro a model to generate acidosis by CO2 bubbling "organ chambers", which would be useful for researchers that aim to study the effects of acid-base disturbs on the endothelium-dependent vascular reactivity. Eighteen male Wistar rats (230-280 g) were housed, before the experiments, under standard laboratory conditions (12h light/dark cycle at 21°C), with free access to food and water. The protocol for promoting in vitro respiratory acidosis was carried out by bubbling increased concentrations of CO2. The target was to achieve an ideal way to decrease the pH gradually to a value of approximately 6.6.It was used, initially, a gas blender varying concentrations of the carbogenic mixture (95% O2 + 5% CO2) and pure CO2. 1) 100% CO2, pH variation very fast, pH minimum 6.0; 2) 90%CO2 pH variation bit slower, pH minimum 6.31; 3) 70%CO2, pH variation slower, pH minimum 6.32; 4) 50% CO2, pH variation slower, pH minimum 6:42; 5) 40 %CO2, Adequate record, pH minimum 6.61, and; 6) 30 %CO2 could not reach values below pH minimum 7.03. Based on these data the gas mixture (O2 60% + CO2 40%) was adopted. This gas mixture (O2 60% + CO2 40%) was effective in inducing respiratory acidosis at a speed that made, possible the recording of isometric force.
Highlights
Acid-base shifts are caused by pCO2 changes and addition of acid or base.These shifts affect the vascular reactivity, and, besides the direct effect on vascular tone, may alter vascular vasoconstrictor and/or vasodilatorsresponsiveness[1,2,3].This study proposes to develop a method in which acidosis is induced in vitro by bubbling CO2 in Krebs solution (“organ chambers” bath), in adequate interval of time to record the changes in isometric force
The present study was carried out to create an in vitro model to generate acidosis by CO2 bubbling “organ chambers”, which would be useful for researchers that aim to study the effects of acid-base disturbs on the endothelium-dependent vascular reactivity
The mechanisms by which pH influences vascular tone or their response to specific agonists are not yet fully understood, but there is some evidence to suggest the involvement of nitric oxide (NO), prostacyclin (PGI2), channels for potassium and calcium flux[3]
Summary
Acid-base shifts are caused by pCO2 changes (respiratory mechanisms) and addition of acid or base (non-respiratory mechanisms).These shifts affect the vascular reactivity, and, besides the direct effect on vascular tone, may alter vascular vasoconstrictor and/or vasodilatorsresponsiveness[1,2,3]. This study proposes to develop a method in which acidosis is induced in vitro by bubbling CO2 in Krebs solution (“organ chambers” bath), in adequate interval of time to record the changes in isometric force. The present study was carried out to create an in vitro model to generate acidosis by CO2 bubbling “organ chambers”, which would be useful for researchers that aim to study the effects of acid-base disturbs on the endothelium-dependent vascular reactivity
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