Iodixanol Induces Apoptotic and Antiproliferative Effects but no Necrotic Cell Death in Renal Proximal Tubular Cells In Vitro

 

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Zusammenfassung

Ziel: Evaluation zytotoxischer Effekte des iso-osmolaren Kontrastmittels Iodixanol an renalen Tubuluszellkulturen. Material und Methoden: LLC-PK1-Zellen wurden mit Iodixanol bzw. isotoner NaCl-Lösung inkubiert (18,75 – 75 mg I/ml, 1 – 24 h). Zelltod wurde durch den Trypan-blau-Ausschluss-Test erfasst. Als Hinweis auf Apoptose wurden Mono- und Oligonukleosomen im Zell-Lysat bestimmt. Messungen der BrdU (5-bromo-2′deoxyuridine)-Inkorporation in zelluläre DNA wurden zur Quantifizierung von Zellproliferation verwendet. Ergebnisse: Iodixanol induzierte keinen signifikanten Anstieg der Zahl nekrotischer Zellen (8 bzw. 9 % bei 37,5 bzw. 75 mg I/ml vs. 8 % in der Kontrolle, p > 0,05). Im Gegensatz dazu erhöhte Iodixanol signifikant die Anzahl der Oligonukleosomen als Hinweis auf Induktion von Apoptose (125 ± 4 % der Kontrolle, p < 0,05). Iodixanol induzierte eine signifikante, dosis- und zeitabhängige Hemmung der BrdU-Inkorporation als Hinweis auf eine Hemmung der Zellproliferation (92 ± 2 % bzw. 79 ± 2 % der Kontrolle bei 18,75 bzw. 37,5 mg I/ml, p < 0,001). Schlussfolgerung: Apoptose und antiproliferative Effekte von Iodixanol, aber nicht die Induktion von Zellnekrose trägt zur renalen Tubulustoxizität von Iodixanol bei.

Abstract

Purpose: To evaluate the cytotoxic effects of the iso-osmolar contrast medium iodixanol on renal tubular cell cultures. Materials and Methods: LLC-PK1 cells were incubated with iodixanol and isotonic NaCl (18.75 – 75 mg I/ml, 1 – 24 hours). Cell death was assessed by the trypan blue exclusion test. To assess apoptosis, mononucleosomes and oligonucleosomes of cell lysates were determined. Measurement of BrdU (5-bromo-2’-deoxyuridine) incorporation into the DNA was used for the quantification of cell proliferation. Results: Iodixanol did not induce any significant increase in the number of necrotic cells (8 % and 9 % at 37.5 and 75 mg I/ml vs. 8 % for control, p > 0.05). In contrast, iodixanol significantly increased the number of oligonucleosomes indicating induction of apoptosis (125 ± 4 % of control, p < 0.05). Iodixanol induced a significant, dose- and time-dependent inhibition of BrdU incorporation indicating the inhibition of cell proliferation (92 ± 2 % and 79 ± 2 % of control at 18.75 and 37.5 mg I/ml, p < 0.001). Conclusion: Apoptosis plus an antiproliferative effect of iodixanol without cell necrosis contribute to the renal tubular toxicity of iodixanol.

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Dr. Marc C. Heinrich

Radiologisches Institut, Universitätsklinikum Erlangen

Maximiliansplatz 1

91054 Erlangen

Germany

Phone: + + 49/91 31/8 53 60 66

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