Using the phenylalanine (PHE) stable isotope pulse method to measure intracellular protein breakdown and metabolic shunting in the context of sepsis in the pig
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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 primedcontinuous 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 WbRaPULSE with L[15N]PHE. Plasma PHE enrichment by LCMS, WbRaCONTI = I/TTR, compartmental analysis (SAAM II) PULSE. Data (mean (SE)); stats (Prism).WbRaCONTI was 22% higher in septic pigs {924 (23) to 1130 (46) nmol/kg BW/min} (p=0.0032), while WbRaPULSE 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}. RaPULSE relates to intracellular PB (p<0.0001) and RaCONTI to RaPULSE (P<0.001).Changes in RaCONTI 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.
