An in vitro evaluation of the glycation potential of a natural disaccharide, trehalose.
BACKGROUND
Glucose, an osmotic agent generally used in continuous ambulatory peritoneal dialysis (CAPD) dialysate, has a critical characteristic of forming advanced glycation endproducts (AGEs). We undertook this study to investigate whether a possible osmotic agent, trehalose, formed fewer AGEs than glucose.


METHODS
Hemoglobin (Hb), a counter-protein of AGE, was incubated in four kinds of medium; glucose-phosphate buffered saline (PBS), autoclaved glucose-PBS, trehalose-PBS, and autoclaved trehalose-PBS, for 3, 7, 14, and 30 days, respectively. Polymerization of the Hb molecule was detected by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and carboxymethylated Hb was detected by Western blotting, using specific mono-clonal antibody for carboxymethylated N-terminal valine-Hb (CMV-Hb).


RESULTS
PBS containing glucose showed bands of polymerized Hb molecule, a phenomenon which was markedly exaggerated by autoclaving. Likely, PBS containing glucose showed the formation of CMV-Hb in the long incubation of 30 days, and PBS containing autoclaved glucose showed accelerated formation of CMV-Hb in an incubation as short as 3 days. By contrast, PBS containing trehalose showed much less increase in a band of 30 k Dalton and in CMV-Hb formation even in autoclaved medium.


CONCLUSIONS
Our present in vitro study clearly showed the superior characteristic of trehalose to produce fewer AGEs. Based upon the results of this study, we propose that the application of trehalose should be considered for CAPD solution.
Motomiya
Yoshihiro
Y
Suiyukai Clinic, 676-1 Kuzumoto-cho, Kashihara, Nara 634-0007, Japan. motomiya@silver.ocn.ne.jp
Higashi
Takuya
T
Masuda
Mari
M
Iwamoto
Hisahiko
H
Miura
Keisuke
K
Yoshimura
Yoshimichi
Y
Maruyama
Ikuro
I
D000818:Animals; D015153:Blotting, Western; D015314:Dialysis Solutions / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006442:Glycated Hemoglobin / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000378:metabolism; D006031:Glycosylation; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D010531:Peritoneal Dialysis, Continuous Ambulatory; D014199:Trehalose / Q000378:metabolism*
D015314:Dialysis Solutions; D006442:Glycated Hemoglobin; D017127:Glycation End Products, Advanced; D006454:Hemoglobins; D014199:Trehalose