title: Folic acid supplementation does not reduce intracellular homocysteine, and may disturb intracellular one-carbon metabolism.
abstract: BACKGROUND
  In randomized trails, folic acid (FA) lowered plasma homocysteine, but failed to reduce cardiovascular risk. We hypothesize this is due to a discrepancy between plasma and intracellular effects of FA.
  
  
  METHODS
  In a double-blind trial, 50 volunteers were randomized to received 500 µg FA daily for 8 weeks, or placebo. Plasma and peripheral blood mononuclear cell (PBMC) concentrations of homocysteine, S-adenosylmethionine (SAM), S-adenosylhomocysteine, methionine, cystathionine and 5-methyltetrahydrofolate (bioactive folate) were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). PBMCs were used as a cellular model since they display the full spectrum of one-carbon (1C) enzymes and reactions.
  
  
  RESULTS
  At baseline, plasma concentrations were a poor reflection of intracellular concentrations for most 1C metabolites, except 5-methyltetrahydrofolate (R=0.33, p=0.02), homocysteine (Hcy) (R=0.35, p=0.01), and cystathionine (R=0.45, p=0.001). FA significantly lowered plasma homocysteine (p=0.00), but failed to lower intracellular homocysteine or change the concentrations of any of the other PBMC 1C metabolites. At baseline, PBMC homocysteine concentrations correlated to PBMC SAM. After FA supplementation, PBMC homocysteine no longer correlated with PBMC SAM, suggesting a loss of SAM's regulatory function. In vitro experiments in lymphoblasts confirmed that at higher folate substrate concentrations, physiological concentrations of SAM no longer effectively inhibit the key regulatory enzyme methylenetetrahydrofolate reductase (MTHFR).
  
  
  CONCLUSIONS
  FA supplementation does not reduce intracellular concentrations of Hcy or any of its closely related substances. Rather, FA may disturb physiological regulation of intracellular 1C metabolism by interfering with SAM's inhibitory effect on MTHFR activity.
authors.affiliation: Department Clinical Chemistry, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Desirée E C
authors.initials: DE
authors.lastname: Smith
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Hornstra
authors.forename: Robert M
authors.initials: RM
authors.lastname: Kok
authors.forename: Henk J
authors.initials: HJ
authors.lastname: Blom
authors.forename: Yvo M
authors.initials: YM
authors.lastname: Smulders
mesh_terms: D000328:Adult; D002244:Carbon / Q000378:metabolism*; D019587:Dietary Supplements; D004311:Double-Blind Method; D005260:Female; D005492:Folic Acid / Q000008:administration & dosage / Q000097:blood; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D012016:Reference Values; D055815:Young Adult
chemical_list: D006710:Homocysteine; D002244:Carbon; D005492:Folic Acid