Lactic acidosis is a frequent cause of poor outcome in the intensive care settings. We set up an experimental model of lactic acid infusion in normoxic and normotensive rats to investigate the systemic effects of lactic acidemia per se without the confounding factor of an underlying organic cause of acidosis.
Sprague Dawley rats underwent a primed endovenous infusion of L(+) lactic acid during general anesthesia. Normoxic and normotensive animals were then randomized to the following study groups (n = 8 per group): S) sustained infusion of lactic acid, S+B) sustained infusion+sodium bicarbonate, T) transient infusion, T+B transient infusion+sodium bicarbonate. Hemodynamic, respiratory and acid-base parameters were measured over time. Lactate pharmacokinetics and muscle phosphofructokinase enzyme's activity were also measured.
Following lactic acid infusion blood lactate rose (P<0.05), pH (P<0.05) and strong ion difference (P<0.05) drop. Some rats developed hemodynamic instability during the primed infusion of lactic acid. In the normoxic and normotensive animals bicarbonate treatment normalized pH during sustained infusion of lactic acid (from 7.22±0.02 to 7.36±0.04, P<0.05) while overshoot to alkalemic values when the infusion was transient (from 7.24±0.01 to 7.53±0.03, P<0.05). When acid load was interrupted bicarbonate infusion affected lactate wash-out kinetics (P<0.05) so that blood lactate was higher (2.9±1 mmol/l vs. 1.0±0.2, P<0.05, group T vs. T+B respectively). The activity of phosphofructokinase enzyme was correlated with blood pH (R2 = 0.475, P<0.05).
pH decreased with acid infusion and rose with bicarbonate administration but the effects of bicarbonate infusion on pH differed under a persistent or transient acid load. Alkalization affected the rate of lactate disposal during the transient acid load.
Citation: Valenza F, Pizzocri M, Salice V, Chevallard G, Fossali T, et al. (2012) Sodium Bicarbonate Treatment during Transient or Sustained Lactic Acidemia in Normoxic and Normotensive Rats. PLoS ONE 7(9): e46035. doi:10.1371/journal.pone.0046035
Editor: Olivier Kocher, Harvard Medical School, United States of America
Received: May 23, 2012; Accepted: August 28, 2012; Published: September 28, 2012
Copyright: © Valenza et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: The study was funded by Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico, Milan, Italy. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Whether it is worth to correct acidemia by infusion of alkaline solutions is a matter of discussion –. The evidence in favour of pH correction of organic acidemia is poor. Clinical studies are few and inconclusive, particularly with respect to clinical outcome ; . There are a number of evidences against alkalinization therapy –. International guidelines “recommend against the use of sodium bicarbonate therapy for the purpose of improving hemodynamics or reducing vasopressor requirements in [septic] patients with hypoperfusion-induced lactic acidemia with pH>7.15” . However, the correction of acidemia is common in clinical practice. An on line survey has recently shown that 67% of critical care physicians start to administer alkaline solutions to patients with lactic acidosis when pH is 7.2 .
Many experimental models of lactic acidemia do not allow to investigate the effects of a lactic acid load per se, because of confounding factors. Moving from the consideration that in clinical setting lactic acidosis may be transient, as during reperfusion of ischemic regions, or sustained, as in persistent hemodynamic instability, we decided to investigate the effects of bicarbonate infusion in normoxic and normotensive animals subjected to transient or sustained lactic acid load.
We wish to report here the results of our investigation and discuss the possible underlying mechanisms and clinical implications.
Materials and Methods
This experimental study was performed after the Ethics Committee of our institution and the Italian Ministry of Health approved the protocol (Permit Number: 6/07). All surgery was performed under anesthesia, and all efforts were made to minimize suffering.
A schematic overview of the experiment flow is shown in Figure 1.
Figure 1. Protocol overview.
A schematic overview of the experiment flow is shown in the figure. The investigation consisted of an initial lactic acid load to induce lactic acidemia followed by randomization to sustained (S) or transient (T) lactic acid infusion with or without sodium bicarbonate (B) treatment.doi:10.1371/journal.pone.0046035.g001
The investigation consisted of an initial lactic acid load to induce lactic acidemia followed by randomization to sustained or transient lactic acid infusion with or without sodium bicarbonate treatment.
Anaesthesia and animal preparation
Sprague Dawley rats (weight 250–300 grams) purchased from Charles River, housed in a warmed and humidified ambient with a 12/12 hours day/night shift, received an intraperitoneal injection of 80 mg/kg thiopental. The trachea was cannulated with a 14 gauge tube connected to a pressure transducer (Motorola MPX 2010DP, Phoenix, AZ, USA). Paralysis was obtained with vecuronium bromide 3 mg/kg i.v. Right carotid artery, femoral and subclavian vein were cannulated with 22 gauge catheters. The arterial catheter was connected to a pressure transducer (Bentley Trantec 800, Santa Ana, CA, USA). Blood pressure and airway pressure were continuously monitored and digitally stored (Elekton Colligo, Agliano, AT, Italy) for subsequent analysis.
During the surgical preparation, rats were mechanically ventilated (Harvard-Rodent 683, Harvard Apparatus South Natick, Massachusetts, USA), with a tidal volume of 6 ml/kg, PEEP of 3 cmH2O and respiratory rate set according to mixed expired CO2 (mixCO2), continuously analyzed (Ohmeda 5250 RGM, Ohmeda, Louisville, CO, USA). At the end of the procedure blood was drawn from the arterial line for blood gas analysis (1620 pH/Blood Gas Analyzer and 682 CO-Oxymeter, Instrumentation Laboratory, Lexington, MA, USA). Respiratory rate was set to obtain the desired values of PaCO2 and pH. Inclusion criteria were: pH 7.35–7.45, PaCO2 35–45 mmHg, lactate <2 mmol/l, hemoglobin >12 g/dl, rectal temperature >36°C, mean arterial pressure (MAP) >90 mmHg. Ventilator parameters remained unchanged throughout the protocol.
If no major problems occurred during surgical preparation and after stabilization time animals were included into the study.
Lactic acid load and randomization process
After confirming inclusion criteria, 14.45 mmol/kg of a 0.55 M solution of L(+) lactic acid (30% in H2O by weight - CH3CH(OH)CO2H- Sigma Aldrich) was infused over 75 minutes through a catheter positioned in a central vein. Animals that met inclusion criteria (pH<7.3, lactate >3 mmol/l and mean arterial pressure >70 mmHg) after the acid load, were randomized by sealed envelopes to one of the following treatments (n = 8 animals per group): S) sustained infusion of lactic acid, S+B) sustained infusion+sodium bicarbonate, T) transient infusion of lactic acid, T+B) transient infusion+sodium bicarbonate. In the sustained groups (group S and S+B), lactic acid was infused throughout the protocol at a rate of 0.20 mmol/kg/min. In the transient groups (groups T and T+B), an equal amount of normal saline was infused. In animals randomized to bicarbonate infusion (groups S+B and T+B) a 1 M solution of sodium bicarbonate was infused at a rate of 0.137 mmol/kg/min; bicarbonate infusion rate was chosen according to pilot studies that suggested a lactate to bicarbonate infusion ratio of 1 M : 0.7 M. If bicarbonate was not infused (groups S and T) an equal volume of normal saline was given to the animals. Experiments were interrupted 60 minutes after randomization (135 minutes after the beginning of the infusion of acid lactic) or if animals developed severe and fatal hypotension.
Acid base parameters included measured (pH, pCO2), and calculated (HCO3, BE) variables (1620 pH/Blood Gas Analyzer, Instrumentation Laboratory, Lexington, MA, USA). Lactate (Lac), sodium (Na), potassium (K), chloride (Cl) and ionized calcium (iCa) ions were measured (ABL555, Radiometer Danmark) and apparent Strong Ion Difference (SIDa) was calculated as
Glucose and hemoglobin concentration were also measured (ABL555, Radiometer Danmark).
Hemodynamic and respiratory parameters.
Hemodynamic parameters (arterial blood pressure and heart rate) and ventilator settings (respiratory rate, tidal volume, positive end-expiratory pressure, mean airway pressure) were recorded throughout the protocol. Oxygenation was studied by arterial blood gas analysis (1620 pH/Blood Gas Analyzer and 682 CO-Oxymeter, Instrumentation Laboratory, Lexington, MA, USA).
Lactate wash-out kinetics was studied in the animals where lactate infusion was interrupted after randomization (transient groups: group T and group T+B).
Lactate kinetics was studied using a model previously described . On the base of lactate increase over the first 75 minutes of infusion, clearance of exogenous lactate and basal lactate production were calculated. Lactate clearance (ml/kg/min) was calculated as the ratio between the lactate load (mmol/kg) and the area under the lactate concentration curve over time (mmol/min/l). We considered endogenous lactate production constant over the first 75 minutes of lactate infusion and we calculated basal lactate production (µmol/kg/min) as the product of basal lactate concentration (Lacbasal - time 0) and exogenous Lac clearance.
Lactate pharmacokinetics was assessed using the model:
fitting, animal per animal, lactate concentrations (y) and time (x), where time 0 was considered time 75. The fitting was performed by means of the least squares method using Sigma Stat software (Systat Software, Inc.). From the fitting analysis coefficient b and c were derived animal by animal; using coefficient b, half time decay (T½) was calculated as 1/b. Coefficients and time decay were then compared for statistical significance.
Phosphofructokinase (PFK) activity.
After the analysis of the first 32 randomized animals, we conducted a new set of experiments to better interpret our results. Eleven animals were randomized to receive the lactic acid load as previously described. Three animals were sacrificed soon after the acid load; 4 animals received transient infusion of lactic acid+sodium bicarbonate infusion (group T+B) and 4 the acid load without sodium bicarbonate infusion (group T). Oxidative soleus muscle (MS) and glycolytic extensor digitorum longus (ME) were then collected and stored with snap freezing technique for PFK activity analysis. Enzymes' activities were also determined . Protein concentration was measured according to Lowry , and the PFK enzymatic activity was expressed as µmol/min/mg of protein.
Results are expressed as mean ± SEM. Analysis of variance was conducted and Bonferroni test was used for all pair-wise comparisons, when indicated. To compare two groups of variables T-test was used or Mann-Whitney Rank Sum Test if normality test failed. Least square linear regression analysis was used to correlate variables. Multiple linear regression was used to correlate base excess and lactate, animal by animal. Statistical significance was accepted as P<0.05. Analysis was performed with the SAS System for Windows version 9.1, unless otherwise specified.
Effects of lactic acid load
The infusion of lactic acid caused lactate to rise (1.2±0.07 mmol/l to 5.5±0.23, P<0.05 time 0 vs. 75, Figure 2) and pH to drop (7.426±0.005 vs. 7.227±0.009, P<0.05, Figure 3). SIDa decreased (31.92±0.54 mEq/l vs. 25.27±0.88, P<0.05) and hemoglobin was significantly lower (13.5±0.3 mg/dl vs. 10.9±0.3, P<0.05). Hyperchloremia (107±0.6 mEq/l vs. 112±0.7, P<0.05) and hypercapnia (40.2±0.6 mmHg vs. 51.2±1.5, P<0.05) also developed. Base excess decreased from 2.08±0.5 mmol/l to −6.3±0.8 (P<0.05). Changes of base excess correlated with changes of lactate (R2 = 0.81, P<0.05, multiple linear regression). Data are presented in Table 1.
Figure 2. Lactate over time in the four groups.
Lactic acid load caused blood lactate to rise in all groups (# P<0.05 vs. time 0). After 135 minutes in the sustained groups blood lactate remained high both in the sustained (S) and the sustained+NaHCO3 (S+B) group. In the transient (T) groups blood lactate levels after 135 minutes were different from values at time 75 minutes (° P<0.05). Animals that received NaHCO3 (T+B) had higher lactate levels (* P<0.05 vs. transient group).doi:10.1371/journal.pone.0046035.g002
Figure 3. Blood pH over time in the four groups.
After 75 minutes of infusion of lactic acid blood pH drop in all groups (# P<0.05 vs. time 0). At 135 minutes pH normalized in the transient group (T) while overshoot to alkalemic values when animals received NaHCO3 (T+B). In the sustained group (S) pH continued to drop while alkaline infusion (S+B) resulted in correction of acidosis. ° P<0.05 vs time 75; * P<0.05 vs. control.doi:10.1371/journal.pone.0046035.g003
Table 1. Acid base variables and plasma chemistry.doi:10.1371/journal.pone.0046035.t001
Hemodynamic and respiratory parameters.
During the primed infusion of lactic acid 11 animals developed severe hemodynamic instability and were excluded. Three animals died soon after randomization (one in group S, two in group S+B) and were replaced in the randomization process so that a total of 32 normoxic and normotensive rats completed the randomization process (n = 8 per group). At baseline, excluded and randomized animals were similar in terms of weight, surgical time, respiratory, acid base and hemodynamic variables except for a trend towards higher values of lactate (1.23±0.07 mmol/l vs. 1.49±0.11, P = 0.056 randomized animals vs. animals who failed lactic acid infusion, respectively), and a significantly higher heart rate (482±17 vs. 431±8 bpm, P<0.05).
Effects of randomization
After the randomization, pH drop over time during the sustained infusion of lactic acid. There was a non significant increase of lactate levels. In the transient groups (T and T+B) 15 minutes after the end of lactic acid infusion, blood lactate concentration, pH and BE values were normal.
When NaHCO3 was infused, pH normalized in the group with sustained infusion (group S+B: from 7.22±0.02 to 7.36±0.04, P<0.05) while rose to alkalemic values in the transient group (group T+B: from 7.24±0.01 to 7.53±0.03, P<0.05). Effects on pH were mainly related to sodium dependent changes of SIDa: Na+ increased from 136.2±3.3 mEq/l to 144.3±1.1 and from 137.5±1.1 to 147.8±1.2 in the sustained group (S+B, P<0.05) and in the transient group (T, P<0.05), respectively.
Hemodynamic and respiratory parameters.
As shown in Table 2, through the experimental time, mean arterial pressure and heart rate were similar in sustained and transient lactic acidosis groups. Sodium bicarbonate infusion did not modify hemodynamic parameters.
Table 2. Hemodynamic and respiratory variables.doi:10.1371/journal.pone.0046035.t002
Ventilator parameters were unchanged and were similar among randomization groups. At the end of the 135 minutes, animals that received sodium bicarbonate infusion were more hypoxemic then their relative controls: 216±36.12 mmHg vs. 269±31.4, group S+B vs group S (P = 0.0511) and 272±16.96 mmHg vs. 332±15.87, group T+B vs group T (P<0.05).
As shown in Figure 2, at the end of the protocol lactate was significantly higher in the T+B group than in the T group.(2.9±1 mmol/l vs. 1±0.2, group T+B vs. group T respectively, P<0.05 – Table 1). Despite similar endogenous lactate production and clearance, the decay of lactate over time was different when bicarbonate was added (Table 3).
Table 3. Lactate pharmacokinetics.doi:10.1371/journal.pone.0046035.t003
When sodium bicarbonate was infused (group T+B) blood glucose concentration slightly decreases over time (112±8 mg/dl vs. 79±13, time 75 vs. 135 respectively, P = 0.095 – Table 1). Glucose and lactate decay was different in T and T+B groups (Figure 4). Glucose changes over time inversely correlated with changes of lactate (R2 = 0.582, P<0.05): the higher the changes of lactate, the lower the glucose changes.
Figure 4. Blood lactate and blood glucose decrease in transient lactic acid infusion group.
Decrease over time of blood lactate concentration and glycaemia in the animals with transient lactic acid infusion. Values are expressed as percentage decrease from time 75 to 135 (*P<0.05 vs. transient group).doi:10.1371/journal.pone.0046035.g004
The activity of PFK in the oxidative soleus muscle was similar in the studied groups (1.675±0.171 µmol/min/mg of proteins, P = 0.11). Conversely, PFK activity in the glycolytic extensor digitorum longus muscle was higher when bicarbonate was infused (P = 0.067, Figure 5). The higher the blood pH measured before muscle harvest the higher the activity of PFK, as shown by linear regression analysis (R2 = 0.475, P<0.05).
Figure 5. PFK activity.
Activity of the glycolytic enzyme phosphofructokinase (PFK) in the glycolitic muscle (ME- extensor digitorum longus) was slightly higher when bicarbonate was infused (P = 0.067). Ctrl = end of lactic acid infusion (i.e 75′ after the start of acid load).doi:10.1371/journal.pone.0046035.g005
We set up an experimental model of transient versus sustained lactic acid infusion in normoxic and normotensive rats to investigate the systemic effects of lactic acidemia per se without the confounding factor of an underlying organic acidosis. The load of lactic acid caused acidemia of both metabolic and respiratory origin. These effects quickly reversed during the transient infusion. Bicarbonate treatment allowed to normalize acid-base parameters during the sustained infusion of lactic acid, but led to overshoot alkalization during the transient load of acid that affected lactate washout kinetics and glucose metabolism.
A number of animal experiments have been conducted to investigate lactic acidosis. As opposed to other more clinically relevant models such as hypoxia –; , sepsis ; , hemorrhage , phenformin intoxication ;  or hepatectomy , in this study we used an infusion of lactic acid in normoxic and normotensive animals to titrate systemic blood lactate concentrations and pH. While alkali treatment during hypoxia or hemodynamic instability may interfere with the cause and the systemic effects of the underlying acidemia, our model allowed to investigate changes of pH and lactate kinetics without possible confounding variables such as oxygen delivery impairment, mitochondrial defects or abnormalities of lactate clearance.
The effects of lactic acid load in our experiments were straightforward: pH consistently drop down to less than 7.2, while lactate rose to clinically significant levels. The acidemia that developed was of mixed origin: despite hemodiluition and hyperchloremia, pH changes were mainly due to lactate, as shown by the correlation between base excess drop and lactate rise. However a respiratory contribution to the drop of pH was also evident, whereas minute ventilation was unchanged throughout the protocol. In line with previous experiences , the induction of lactic acidemia resulted in a degree of mortality rate. However, at the beginning of randomization animals included in the randomization process were normoxic and normotensive.
After the randomization, pH drop over time during the sustained infusion of lactic acid. Even if not significant, there was a rise in lactate levels. Since PaCO2 and electrolyte concentrations did not change through the randomization, lactate increase may be interpreted as an extra load of organic acid.
Sodium bicarbonate treatment caused pH to rise. Alkalinization mainly occurred because of a sodium dependent change of SIDa, with some possible contribution of the reduction of weak acids due to hemodilution. The role of respiratory alkalosis was negligible, consistent with the fact that the release of CO2 is known to occur early after the infusion of sodium bicarbonate and depends on both the infusion rate and the concentration of non-bicarbonate buffers – that was relatively low during the randomization time.
Although the starting concentration of bicarbonate was similar in S+B and T+B groups, pH normalized during sustained infusions of lactic acid (from 7.22±0.02 to 7.36±0.04) while increased up to alkalemic values (from 7.24±0.01 to 7.53±0.03) when acid latic infusion was transient. Re-perfusion of hypoperfused or ischemic territories is characterized by an organic acid washout and a transient acid load, that cause a transient pH decrease. In this case sodium bicarbonate infusion may exceed the desired effect of reversing acidemia. On the contrary, the normalization of pH during sustained lactic acid infusion seems to be relevant, even if we did not find hemodynamic instability at low pH that many physicians advocate to start alkalinization therapy .
As expected, sodium significantly increased in both groups treated with bicarbonate. We also observed in these groups an oxygenation decrease probably due to a fluid load.
Bicarbonate infusion affected blood lactate levels differently during sustained or transient acidemia. In fact, lactate slightly rose in the S+B and S groups, but in a similar fashion. On the contrary, when the acid load was transient, at the end of the experiment lactate was significantly higher in the group of animals that received bicarbonate treatment (T+B). The abrupt and wide change of pH that followed bicarbonate infusion in this group possibly affected lactate metabolism, given the modulatory role of pH on blood levels of lactate –; ; . Pharmacokinetic results suggest a reduction of the oxidation of lactate after bicarbonate infusion, according to Chiolerò et al. . Lactate kinetics are also in line with those from Druml et al. who found that respiratory alkalosis decreases the clearance of infused lactic acid , and Abu Romeh et al. who found in a rat model of hypoxic lactic acidemia that systemic acidosis inhibits net lactic acid production.  Because it is known that pH modulates both glycolitic flow – and lactate cellular uptake –, and because lactate undergoes preferential oxydation when in excess , we speculate that lactate was preferentially oxidated at low pH. On the contrary, when bicarbonate was infused, alkalosis favored glucose metabolism so that glucose levels decreased more than lactate and lactate half-life increased. The data on PFK activity seem to confirm this hypothesis.
Provided the effects of bicarbonate infusion on pH differed under a persistent or transient acid load and alkalization affected the rate of lactate disposal during the transient acid load, when deciding to infuse sodium bicarbonate one should take into consideration the metabolic effects of pH on the cell and the possible consequences on adaptation to energy failure ; –.
Conceived and designed the experiments: FV LG. Performed the experiments: MP VS G. Chevallard TF SC SF SG. Analyzed the data: FP FF G. Comi. Contributed reagents/materials/analysis tools: FF G. Comi. Wrote the paper: FV MP VS SC SF LG.
- 1. Broder G, Weil M (1964) Excess lactate: an index of reversibility in human patients. Science 143(3613):1457–1459. doi: 10.1126/science.143.3613.1457
- 2. Weil M, Afifi A (1970) Experimental and clinical studies on lactate and pyruvate as indicators of the severity of acute circulatory failure (shock). Circulation 41(6):989–1001. doi: 10.1161/01.cir.41.6.989
- 3. Vitek V, Cowley RA (1971) Blood lactate in the prognosis of various forms of shock. Ann Surg 173(2):308–313. doi: 10.1097/00000658-197102000-00021
- 4. Cady LD, Weil MH, Afifi A, Michaels SF, Liu VY, et al. (1973) Quantitation of severity of critical illness with special reference to blood lactate. Crit Care Med 1(2):75–80. doi: 10.1097/00003246-197303000-00003
- 5. Bakker J, Coffernils M, Leon M, Gris P, Vincent JL (1991) Blood lactate levels are superior to oxygen-derived variables in predicting outcome in human septic shock. Chest 99(4):956–962. doi: 10.1378/chest.99.4.956
- 6. Gunnerson KJ, Saul M, He S, Kellum JA (2006) Lactate versus non-lactate metabolic acidosis: a retrospective outcome evaluation of critically ill patients. Crit Care 10(1):R22.
- 7. Jansen TC, van Bommel J, Mulder PG, Rommes JH, Schieveld SJM, et al. (2008) The prognostic value of blood lactate levels relative to that of vital signs in the pre-hospital setting: a pilot study. Crit Care 12(6):R160. doi: 10.1186/cc7159
- 8. Khosravani H, Shahpori R, Stelfox HT, Kirkpatrick AW, Laupland KB (2009) Occurrence and adverse effect on outcome of hyperlactatemia in the critically ill. Crit Care 13(3):R90. doi: 10.1186/cc7918
- 9. Nichol AD, Egi M, Pettila V, Bellomo R, French C, et al. (2010) Relative hyperlactatemia and hospital mortality in critically ill patients: a retrospective multi-centre study. Crit Care 14(1):R25. doi: 10.1186/cc8888
- 10. Forsythe SM, Schmidt GA (2000) Sodium bicarbonate for the treatment of lactic acidosis. Chest 117(1):260–267. doi: 10.1378/chest.117.1.260
- 11. Rachoin JS, Weisberg , McFadden CB (2010) Treatment of lactic acidosis: appropriate confusion. J Hosp Med 5(4):E1–E7. doi: 10.1002/jhm.600
- 12. Arieff AI (1996) Current concepts in acid-base balance: use of bicarbonate in patients with metabolic acidosis. Current Anaesthesia & Critical Care 7: 182–186. doi: 10.1016/s0953-7112(96)80091-3
- 13. Mathieu D, Neviere R, Billard V, Fleyfel M, Wattel F (1991) Effects of bicarbonate therapy on hemodynamics and tissue oxygenation in patients with lactic-acidosis: a prospective, controlled clinical study. Crit Care Med 19(11):1352–1356. doi: 10.1097/00003246-199111000-00008
- 14. Cooper DJ, Walley KR, Wiggs BR, Russell JA (1990) Bicarbonate does not improve hemodynamics in critically ill patients who have lactic acidosis. A prospective, controlled clinical study. Ann Intern Med 112(7):492–498. doi: 10.1097/00132586-199012000-00067
- 15. Arieff AI, Leach W, Park R, Lazarowitz VC (1982) Systemic effects of NaHCO3 in experimental lactic acidosis in dogs. Am J Physiol 242(6):F586–F591.
- 16. Graf H, Leach W, Arieff AI (1985) Metabolic effects of sodium bicarbonate in hypoxic lactic acidosis in dogs. Am J Physiol 249(5Pt2):F630–F635.
- 17. Graf H, Leach W, Arieff AI (1985) Evidence for a detrimental effect of bicarbonate therapy in hypoxic lactic acidosis. Science 227(4688):754–756. doi: 10.1126/science.3969564
- 18. Rhee KH, Toro LO, McDonald GG, Nunnally RL, Levin DL (1993) Carbicarb, sodium bicarbonate, and sodium chloride in hypoxic lactic acidosis. Effect on arterial blood gases, lactate concentrations, hemodynamic variables, and myocardial intracellular pH. Chest 104(3):913–918. doi: 10.1378/chest.104.3.913
- 19. Cooper DJ, Herbertson MJ, Werner HA, Walley KR (1993) Bicarbonate does not increase left ventricular contractility during L-lactic acidemia in pigs. Am Rev Respir Dis 148(2):317–322. doi: 10.1164/ajrccm/148.2.317
- 20. Benjamin E, Oropello JM, Abalos AM, Hannon EM, Wang JK, et al. (1994) Effects of acid-base correction on hemodynamics, oxygen dynamics, and resuscitability in severe canine hemorrhagic shock. Crit Care Med 22(10):1616–1623. doi: 10.1097/00003246-199422100-00018
- 21. Tanaka M, Nishikawa T, Mizutani T (1996) Normovolaemic haemodilution attenuates cardiac depression induced by sodium bicarbonate in canine metabolic acidosis. Br J Anaesth 77(3):408–412. doi: 10.1093/bja/77.3.408
- 22. Tanaka M, Nishikawa T (1997) Acute haemodynamic effects of sodium bicarbonate administration in respiratory and metabolic acidosis in anaesthetized dogs. Anaesth Intensive Care 25(6):615–620.
- 23. Boyd JH, Walley KR (2008) Is there a role for sodium bicarbonate in treating lactic acidosis from shock? Curr Opin Crit Care 14(4):379–383. doi: 10.1097/mcc.0b013e3283069d5c
- 24. Dellinger RP, Levy MM, Carlet JM, Bion J, Parker MM, et al. (2008) Surviving Sepsis Campaign: international guidelines for management of severe sepsis and septic shock: 2008. Crit Care Med 36(1):296–327. doi: 10.1007/s00134-008-1090-z
- 25. Kraut JA, Kurtz I (2006) Use of base in the treatment of acute severe organic acidosis by nephrologists and critical care physicians: results of an online survey. Clin Exp Nephrol 10(2):111–117. doi: 10.1007/s10157-006-0408-9
- 26. Chiolero R, Tappy L, Gillet M, Revelly JP, Roth H, et al. (1999) Effect of major hepatectomy on glucose and lactate metabolism. Ann Surg 229(4):505–513. doi: 10.1097/00000658-199904000-00009
- 27. Ling K H, Byrne W, Lardy H (2010) Phoshoexokinase. In: S.P.Colowick and N.0.Kaplan, editors. Methods in enzymology. Academic Press Inc, New York. pp. 306–310.
- 28. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275.
- 29. Abu Romeh S, Tannen RL (1986) Amelioration of hypoxia-induced lactic acidosis by superimposed hypercapnea or hydrochloric acid infusion. Am J Physiol 250(4Pt 2):F702–F709.
- 30. Chrusch C, Bands C, Bose D, Li X, Jacobs H, et al. (2000) Impaired hepatic extraction and increased splanchnic production contribute to lactic acidosis in canine sepsis. Am J Respir Crit Care Med 161(2Pt 1):517–526. doi: 10.1164/ajrccm.161.2.9902403
- 31. Chrusch C, Bautista E, Jacobs HK, Light RB, Bose D, et al. (2002) Blood pH level modulates organ metabolism of lactate in septic shock in dogs. J Crit Care 17(3):188–202. doi: 10.1053/jcrc.2002.35816
- 32. Park R, Arieff AI (1982) Treatment of lactic acidosis with dichloroacetate in dogs. J Clin Invest 70(4):853–862. doi: 10.1172/jci110682
- 33. Gattinoni L, Taccone P, Carlesso E (2006) Respiratory acidosis: is the correction with bicarbonate worth? Minerva Anestesiol 72(6):551–557.
- 34. Levraut J, Garcia P, Giunti C, Ichai C, Bouregba M, et al. (2000) The increase in CO2 production induced by NaHCO3 depends on blood albumin and hemoglobin concentrations. Intensive Care Med 26(5):558–564. doi: 10.1007/s001340051204
- 35. Okamoto H, Hoka S, Kawasaki T, Okuyama T, Takahashi S (1995) Changes in end-tidal carbon dioxide tension following sodium bicarbonate administration: correlation with cardiac output and haemoglobin concentration. Acta Anaesthesiol Scand 39(1):79–84. doi: 10.1111/j.1399-6576.1995.tb05596.x
- 36. Druml W, Grimm G, Laggner AN, Lenz K, Schneeweiss B (1991) Lactic acid kinetics in respiratory alkalosis. Crit Care Med 19(9):1120–1124. doi: 10.1097/00003246-199109000-00005
- 37. Hood VL, Tannen RL (1998) Protection of acid-base balance by pH regulation of acid production. N Engl J Med 339(12):819–826. doi: 10.1056/nejm199809173391207
- 38. Pagliassotti MJ, Donovan CM (1990) Glycogenesis from lactate in rabbit skeletal muscle fiber types. Am J Physiol 258(4Pt 2):R903–R911.
- 39. Miller BF, Fattor JA, Jacobs KA, Horning MA, Navazio F, et al. (2002) Lactate and glucose interactions during rest and exercise in men: effect of exogenous lactate infusion. J Physiol 544(Pt3):963–975. doi: 10.1113/jphysiol.2002.027128
- 40. Hood VL, Tannen RL (1983) Ph control of lactic acid and keto acid production a mechanism of acid-base regulation. Miner Electrolyte Metab 9(4–6):317–325.
- 41. Sestoft L, Marshall MO (1986) Hepatic lactate uptake is enhanced by low pH at low lactate concentrations in perfused rat liver. Clin Sci Lond 70(1):19–22.
- 42. Baron PG, Iles RA, Cohen RD (1978) Effect of varying PCO2 on intracellular pH and lactate consumption in the isolated perfused rat liver. Clin Sci Mol Med 55(2):175–181.
- 43. Roth DA, Brooks GA (1990) Lactate and pyruvate transport is dominated by a pH gradient-sensitive carrier in rat skeletal muscle sarcolemmal vesicles. Arch Biochem Biophys 279(2):386–394. doi: 10.1016/0003-9861(90)90506-t
- 44. Halestrap AP, Price NT (1999) The proton-linked monocarboxylate transporter (MCT) family: structure, function and regulation. Biochem J 343(Pt2):281–299. doi: 10.1042/0264-6021:3430281
- 45. Hollidge-Horvat MG, Parolin ML, Wong D, Jones NL, Heigenhauser GJ (1999) Effect of induced metabolic acidosis on human skeletal muscle metabolism during exercise. Am J Physiol 277(4Pt 1):E647–E658.
- 46. Valenza F, Aletti G, Fossali T, Chevallard G, Sacconi F, et al. (2005) Lactate as a marker of energy failure in critically ill patients: hypothesis. Crit Care 9(6):588–593. doi: 10.1186/cc3818
- 47. Kitano T, Nisimaru N, Shibata E, Iwasaka H, Noguchi T, et al. (2002) Lactate utilization as an energy substrate in ischemic preconditioned rat brain slices. Life Sci 72(4–5):557–564. doi: 10.1016/s0024-3205(02)02251-8
- 48. Pellerin L (2003) Lactate as a pivotal element in neuron-glia metabolic cooperation. Neurochem Int 43(4–5):331–338. doi: 10.1016/s0197-0186(03)00020-2
- 49. Leverve XM (1999) Energy metabolism in critically ill patients: lactate is a major oxidizable substrate. Curr Opin Clin Nutr Metab Care 2(2):165–169. doi: 10.1097/00075197-199903000-00013
- 50. Leverve XM (2001) Inter-organ substrate exchanges in the critically ill. Curr Opin Clin Nutr Metab Care 4(2):137–142. doi: 10.1097/00075197-200103000-00010