Direct inhibitory effect of uremic toxins and polyamines on proliferation of VERO culture cells.
The dialysate fluid of uremic patients exhibits, in vitro, an inhibitory effect on cell growth, owing to urea, guanidino compounds, and substances named middle molecules. The polyamines are compounds which exhibit high levels in biological fluids during either normal development or disease such as psoriasis, uremia, and tumors. Dialysate and middle molecules show toxicity and degeneration of the organotype cultures, whereas the free polyamines and nonrecirculated dialysate do not have any toxic effect. The aim of this study is to analyze the effects of polyamines, nonrecirculated dialysate, and middle molecules of uremic patients in periodic hemodialysis on cultured VERO (fibroblast-like cells) growth. These cells show an inhibition of growth in middle molecules or 2 x 10(-4) M putrescine and a stimulation with nonrecirculated dialysate and 2 x 10(-8) M putrescine. The effect is different because the cultures with middle molecules begin growth again after 24 hr, whereas in the presence of 2 x 10(-4) M putrescine no further growth is observed. Cells maintained in middle molecules + 2 x 10(-8) M putrescine show an irreversible degeneration, attesting a toxic effect due to the low molarities of putrescine. The electron microscopy shows alteration of cytoplasmic, mitochondrial, and nuclear membranes, but no chromatin fragmentation with either middle molecules or 2 x 10(-4) M putrescine: this suggests that the cells do not die of apoptosis. In conclusion, during uremia the polyamines could cause toxic effects, even at low concentrations, on cells stressed by other toxic stimuli.
Stabellini
G
G
Istituto di Istologia ed Embriologia Generale, Università di Ferrara, Italy.
Mariani
G
G
Pezzetti
F
F
Calastrini
C
C
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002522:Chlorocebus aethiops; D004247:DNA / Q000032:analysis; D015314:Dialysis Solutions / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects*; D007425:Intracellular Membranes / Q000187:drug effects / Q000648:ultrastructure; D008854:Microscopy, Electron; D008928:Mitochondria / Q000187:drug effects / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000187:drug effects / Q000648:ultrastructure; D011073:Polyamines / Q000302:isolation & purification / Q000494:pharmacology*; D011700:Putrescine / Q000494:pharmacology; D013997:Time Factors; D014118:Toxins, Biological / Q000302:isolation & purification / Q000494:pharmacology*; D014511:Uremia / Q000378:metabolism*; D014709:Vero Cells
D015314:Dialysis Solutions; D011073:Polyamines; D014118:Toxins, Biological; C026865:uremia middle molecule toxins; D004247:DNA; D011700:Putrescine