D-lactate increases pulmonary apoptosis by restricting phosphorylation of Bad and eNOS in a rat model of hemorrhagic shock

Amín Jaskille, Hasan B. Alam, Peter Rhee, William Hanes, John R. Kirkpatrick, Elena Koustova, Henri Ford, Edwin A. Deitch, Tetsu Yukioka

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Resuscitation with racemic lactated Ringer's solution (containing equal amounts of D and L isomers of lactate) has been shown to induce pulmonary apoptosis. Substitution of DL-isomer lactate with ketone bodies (β-hydroxybutyrate, BHB), sodium pyruvate, or L-isomer of lactate decrease this injury without changing the energy status of the tissues or the expression of apoptotic genes. These modified solutions however alter the function of apoptotic proteins through an unknown mechanism. We postulated that DL-LR induces apoptosis by restricting the phosphorylation of key apoptotic proteins. Methods: Male Sprague Dawley rats (n = 30, 5/group) were subjected to a three stage, volume-controlled hemorrhage and randomized to the following groups. 1) No hemorrhage (Sham); 2) Hemorrhage and no resuscitation (NR); 3) Resuscitation with 3x shed blood volume of racemic LR (DL-LR); 4) Resuscitation with 3x shed blood volume of LR containing only the L-isomer of lactate (L-LR); 5) Resuscitation with 3x shed blood volume of pyruvate Ringer's (PR); 6) Resuscitation with 3x shed blood volume of ketone Ringer's (KR). The modified Ringer's solutions were identical to racemic LR except for equimolar substitution of DL-lactate for L-lactate, pyruvate and BHB respectively. Lung tissue was obtained 2 hours later and subjected to Western Blotting. The levels of Akt, Bad, and eNOS (total and phosphorylated) proteins were measured. Finally, the expression of gene coding for protein 14-3-3 was measured using RT-PCR. Results: Resuscitation with DL-LR caused a significant (p < 0.05) increase in the total Bad and a decrease in phosphorylated Bad protein expression in the lung. It also caused an increase in the phosphorylated Akt levels and a decrease in gene coding for protein 14-3-3. These changes were consistent with signaling imbalances that favor apoptosis. Modified LR solutions, on the other hand, did not cause these alterations. Phosphorylation pattern of eNOS supported the involvement of PI3K/Akt pathway in this process. Conclusions: Racemic lactate plays a role in the induction of pulmonary apoptosis by restricting phosphorylation of Bad and eNOS proteins.

Original languageEnglish (US)
Pages (from-to)262-270
Number of pages9
JournalJournal of Trauma - Injury, Infection and Critical Care
Volume57
Issue number2
DOIs
StatePublished - Aug 1 2004
Externally publishedYes

Fingerprint

Hemorrhagic Shock
Resuscitation
Lactic Acid
Phosphorylation
Apoptosis
Lung
Blood Volume
bcl-Associated Death Protein
Pyruvic Acid
14-3-3 Proteins
Hemorrhage
Proteins
Hydroxybutyrates
Ketone Bodies
Ketones
Phosphatidylinositol 3-Kinases
Sprague Dawley Rats
Western Blotting
Sodium
Gene Expression

Keywords

  • Akt
  • Apoptosis
  • Bad
  • eNOS
  • Hemorrhagic shock
  • Isomers
  • Ketone body
  • Lactated Ringer's
  • Lung
  • Phosphorylation
  • Pyruvate
  • Racemic
  • Resuscitation

ASJC Scopus subject areas

  • Surgery
  • Critical Care and Intensive Care Medicine

Cite this

D-lactate increases pulmonary apoptosis by restricting phosphorylation of Bad and eNOS in a rat model of hemorrhagic shock. / Jaskille, Amín; Alam, Hasan B.; Rhee, Peter; Hanes, William; Kirkpatrick, John R.; Koustova, Elena; Ford, Henri; Deitch, Edwin A.; Yukioka, Tetsu.

In: Journal of Trauma - Injury, Infection and Critical Care, Vol. 57, No. 2, 01.08.2004, p. 262-270.

Research output: Contribution to journalArticle

Jaskille, Amín ; Alam, Hasan B. ; Rhee, Peter ; Hanes, William ; Kirkpatrick, John R. ; Koustova, Elena ; Ford, Henri ; Deitch, Edwin A. ; Yukioka, Tetsu. / D-lactate increases pulmonary apoptosis by restricting phosphorylation of Bad and eNOS in a rat model of hemorrhagic shock. In: Journal of Trauma - Injury, Infection and Critical Care. 2004 ; Vol. 57, No. 2. pp. 262-270.
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abstract = "Resuscitation with racemic lactated Ringer's solution (containing equal amounts of D and L isomers of lactate) has been shown to induce pulmonary apoptosis. Substitution of DL-isomer lactate with ketone bodies (β-hydroxybutyrate, BHB), sodium pyruvate, or L-isomer of lactate decrease this injury without changing the energy status of the tissues or the expression of apoptotic genes. These modified solutions however alter the function of apoptotic proteins through an unknown mechanism. We postulated that DL-LR induces apoptosis by restricting the phosphorylation of key apoptotic proteins. Methods: Male Sprague Dawley rats (n = 30, 5/group) were subjected to a three stage, volume-controlled hemorrhage and randomized to the following groups. 1) No hemorrhage (Sham); 2) Hemorrhage and no resuscitation (NR); 3) Resuscitation with 3x shed blood volume of racemic LR (DL-LR); 4) Resuscitation with 3x shed blood volume of LR containing only the L-isomer of lactate (L-LR); 5) Resuscitation with 3x shed blood volume of pyruvate Ringer's (PR); 6) Resuscitation with 3x shed blood volume of ketone Ringer's (KR). The modified Ringer's solutions were identical to racemic LR except for equimolar substitution of DL-lactate for L-lactate, pyruvate and BHB respectively. Lung tissue was obtained 2 hours later and subjected to Western Blotting. The levels of Akt, Bad, and eNOS (total and phosphorylated) proteins were measured. Finally, the expression of gene coding for protein 14-3-3 was measured using RT-PCR. Results: Resuscitation with DL-LR caused a significant (p < 0.05) increase in the total Bad and a decrease in phosphorylated Bad protein expression in the lung. It also caused an increase in the phosphorylated Akt levels and a decrease in gene coding for protein 14-3-3. These changes were consistent with signaling imbalances that favor apoptosis. Modified LR solutions, on the other hand, did not cause these alterations. Phosphorylation pattern of eNOS supported the involvement of PI3K/Akt pathway in this process. Conclusions: Racemic lactate plays a role in the induction of pulmonary apoptosis by restricting phosphorylation of Bad and eNOS proteins.",
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AU - Jaskille, Amín

AU - Alam, Hasan B.

AU - Rhee, Peter

AU - Hanes, William

AU - Kirkpatrick, John R.

AU - Koustova, Elena

AU - Ford, Henri

AU - Deitch, Edwin A.

AU - Yukioka, Tetsu

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Y1 - 2004/8/1

N2 - Resuscitation with racemic lactated Ringer's solution (containing equal amounts of D and L isomers of lactate) has been shown to induce pulmonary apoptosis. Substitution of DL-isomer lactate with ketone bodies (β-hydroxybutyrate, BHB), sodium pyruvate, or L-isomer of lactate decrease this injury without changing the energy status of the tissues or the expression of apoptotic genes. These modified solutions however alter the function of apoptotic proteins through an unknown mechanism. We postulated that DL-LR induces apoptosis by restricting the phosphorylation of key apoptotic proteins. Methods: Male Sprague Dawley rats (n = 30, 5/group) were subjected to a three stage, volume-controlled hemorrhage and randomized to the following groups. 1) No hemorrhage (Sham); 2) Hemorrhage and no resuscitation (NR); 3) Resuscitation with 3x shed blood volume of racemic LR (DL-LR); 4) Resuscitation with 3x shed blood volume of LR containing only the L-isomer of lactate (L-LR); 5) Resuscitation with 3x shed blood volume of pyruvate Ringer's (PR); 6) Resuscitation with 3x shed blood volume of ketone Ringer's (KR). The modified Ringer's solutions were identical to racemic LR except for equimolar substitution of DL-lactate for L-lactate, pyruvate and BHB respectively. Lung tissue was obtained 2 hours later and subjected to Western Blotting. The levels of Akt, Bad, and eNOS (total and phosphorylated) proteins were measured. Finally, the expression of gene coding for protein 14-3-3 was measured using RT-PCR. Results: Resuscitation with DL-LR caused a significant (p < 0.05) increase in the total Bad and a decrease in phosphorylated Bad protein expression in the lung. It also caused an increase in the phosphorylated Akt levels and a decrease in gene coding for protein 14-3-3. These changes were consistent with signaling imbalances that favor apoptosis. Modified LR solutions, on the other hand, did not cause these alterations. Phosphorylation pattern of eNOS supported the involvement of PI3K/Akt pathway in this process. Conclusions: Racemic lactate plays a role in the induction of pulmonary apoptosis by restricting phosphorylation of Bad and eNOS proteins.

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KW - Phosphorylation

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KW - Racemic

KW - Resuscitation

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