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Using the phenylalanine (PHE) stable isotope pulse method to measure intracellular protein breakdown and metabolic shunting in the context of sepsis in the pig
Author(s) -
Ten Have Gabriella A,
Engelen Marielle P,
Wolfe Robert R,
Deutz Nicolaas E
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.42.1
Subject(s) - intracellular , sepsis , extracellular , pulse (music) , chemistry , shunting , metabolism , medicine , endocrinology , biochemistry , physics , detector , optics
To develop an easy applicable clinical method to measure protein metabolism, we tested the PHE stable isotope pulse method (PULSE) and compared the results with a simultaneously performed primed‐continuous infusion protocol (CONTI) during sepsis in pigs with stimulated whole body protein breakdown (WbRa). 12 pigs with Pseudomonas aeroginosa induced hyperdynamic sepsis and 8 controls were studied 17h after start. WbRa CONTI with L‐[13C6]PHE and WbRa‐PULSE with L‐[15N]PHE. Plasma PHE enrichment by LC‐MS, WbRa‐CONTI = I/TTR, compartmental analysis (SAAM II) PULSE. Data (mean (SE)); stats (Prism). WbRa‐CONTI was 22% higher in septic pigs {924 (23) to 1130 (46) nmol/kg BW/min} (p=0.0032), while WbRa‐PULSE 36% {1722 (66) to 2342 (191); p=0.0164}. Sepsis show a substantial expansion of pool 1 and 2 and metabolic shunting (pool 2><1), while intracellular PB increased 31% {2620 (82) to 3425 (305); p=0.0426}. Ra‐PULSE relates to intracellular PB (p<0.0001) and Ra‐CONTI to Ra‐PULSE (P<0.001). Changes in Ra‐CONTI reflect intracellular PB with high sensitivity. PULSE in addition shows that sepsis in the pig is related to a substantial increase in metabolic shunting in relation to expansion of the pools (probably intra‐ and extracellular). It is suggested that the CONTI method can be replaced with the easy applicable PULSE method that also provides additional information about intracellular fluxes and breakdown.