Document Detail


Histidine inhibits the degradation of cells suspended in Ringer's lactate.
MedLine Citation:
PMID:  18073602     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: We previously demonstrated that the degradation of a suspension of Jurkat cells in Ringer's lactate (RL) was inhibited by the addition of a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid/Tris buffer. Given the ability of histidine to buffer protons in the physiologic range (pKa = 6.0), we hypothesized that this amino acid would have the same effect. METHODS: RL was made in our laboratory using sodium l-lactate. Jurkat cells were suspended in RL alone or RL with various concentrations of histidine or other test reagents at 37 degrees C for 4 hours or 24 hours in an atmosphere of 95% air and 5% CO2. Using flow cytometry, we measured cell shrinkage, phosphatidylserine translocation, propidium iodide uptake, and intracellular oxygen free radical production. RESULTS: Cell shrinkage was induced by suspension in RL after 4 hours incubation. At 4 hours, cell shrinkage was inhibited by all concentrations of histidine tested, 7.8 mumol/L to 10 mmol/L. There was no statistical difference between cells suspended in medium and cells suspended in 1 mmol/L or 10 mmol/L histidine. After 24 hours incubation, 100% of the cells in RL had undergone cell shrinkage whereas in 10 mmol/L histidine only a mean of 20% of the cells had undergone cell shrinkage. The inhibitory effect of 1 mmol/L histidine at pH 7.4 was compared with that at pH 6.8. After 4 hours incubation, there was no difference. After 24 hours incubation, the inhibitory effect at pH 7.4 was significantly greater that that at pH 6.8. Histidine at 1 mmol/L to 10 mmol/L significantly reduced the percentage of cells that underwent phosphatidylserine translocation and propidium iodide uptake. The effect of the dipeptide buffer, glycylglycine, and the two other positively charged amino acids, arginine and lysine, after 4 hours incubation was compared with histidine at 1 mmol/L. At 1 mmol/L, histidine was superior to arginine and lysine and indistinguishable from glycylglycine. Intracellular free radical production was measured at 0.5 mmol/L, 1.0 mmol/L, and 10 mmol/L histidine concentrations. There was significant inhibition only at 10 mmol/L. CONCLUSIONS: Characteristics of apoptotic cell death that occur in cells suspended in RL are inhibited by the addition of histidine, arginine, and lysine as well as the dipeptide glycylglycine, which, with a pKa of 8.25, also buffers in the physiologic range. Histidine is superior to lysine and arginine at 1 mmol/L. The salutary effect of histidine at 0.5 mmol/L and 1 mmol/L is caused by a mechanism other than the inhibition of oxygen free radicals. Moreover, the buffering of protons may play a role at 24 hours but made no difference at 4 hours.
Authors:
Cuthbert O Simpkins; Viktoriya Ekshyyan; Brad Snyder
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of trauma     Volume:  63     ISSN:  1529-8809     ISO Abbreviation:  J Trauma     Publication Date:  2007 Sep 
Date Detail:
Created Date:  2007-12-12     Completed Date:  2008-01-24     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0376373     Medline TA:  J Trauma     Country:  United States    
Other Details:
Languages:  eng     Pagination:  565-72     Citation Subset:  AIM; IM    
Affiliation:
LSU Health Sciences Center, Shreveport, LA, USA. csimpk@lsuhsc.edu
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MeSH Terms
Descriptor/Qualifier:
Analysis of Variance
Apoptosis / drug effects*
Cell Death / drug effects
Cell Membrane Permeability / drug effects
Flow Cytometry
Fluid Therapy*
Free Radicals / metabolism
Histidine / pharmacology*
Humans
Hydrogen-Ion Concentration
Isotonic Solutions / pharmacology*
Jurkat Cells / drug effects*
Phosphatidylserines / metabolism
Chemical
Reg. No./Substance:
0/Free Radicals; 0/Isotonic Solutions; 0/Phosphatidylserines; 71-00-1/Histidine; 8026-10-6/Ringer's solution

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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