The effect of polyamines and dialysate fluid on extracellular matrix synthesis in VERO cell cultures.
BACKGROUND
Polyamines are involved in normal and pathological cell proliferation and differentiation. Like acid radicals, they play an important role in remodelling the extracellular matrix and are considered "uremic toxins". Proteins and glycosaminoglycans are essential components of the extracellular matrix, and contribute to normal mature organ functions. The aim of this study was to analyse the effects of free polyamines, dialysate fluid components and dialysis fluid on protein and extracellular glycosaminoglycan synthesis in VERO cell cultures.


METHODS
The dialysate fluid components were separated with a Sephadex G15 column and the cultures were analysed after incorporation of 3H-leucine and 3H-glucosamine. Cultures were run at pH 7.0 and pH 7.4. The glycosaminoglycan classes were separated with a DEAE column, and polyamines were determined by high-performance liquid chromatography. Proteins and single glycosaminoglycan classes were quantified by a scintillator. DNA gel electrophoresis was done to detect chromatin fragmentation.


RESULTS
Dialysate contained putrescine, spermidine and spermine, chromatography showing four peaks; only peaks I and II indicated polyamines at respectively Da 5000 and 1500. Polyamines are therefore linked to different carriers. There was an increase of protein and glycosaminoglycan synthesis with dialysis fluid and polyamines, but inhibition with peak II or dialysate. DNA gel electrophoresis showed no chromatin fragmentation. Findings at pH 7.0 and 7.4 were similar.


CONCLUSIONS
It would appear that in uremic patients polyamines are conjugated to protein carriers of different molecular weights with different biological actions. As polyamines and dialysis fluid affect changes in extracellular matrix, they could be related to physiological organ functions. However, these in vitro data must be considered with the appropriate limitations when we try to extrapolate them to the in vivo situation.
Stabellini
Giordano
G
Department of Human Anatomy, LITA Segrate, State University of Milan, Italy. giordano.stabellini@unimi.it
Calastrini
Carla
C
Scapoli
Luca
L
Locci
Paola
P
De Mattei
Monica
M
Dellavia
Claudia
C
Moscheni
Claudia
C
Vertemati
Maurizio
M
Bedani
Pier Luigi
PL
D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D002850:Chromatography, Gel; D015314:Dialysis Solutions / Q000494:pharmacology*; D004587:Electrophoresis, Agar Gel; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008875:Middle Aged; D011073:Polyamines / Q000494:pharmacology*; D006435:Renal Dialysis; D012680:Sensitivity and Specificity; D014709:Vero Cells
D015314:Dialysis Solutions; D006025:Glycosaminoglycans; D011073:Polyamines