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The study of cytotoxic effect to creatinine metabolite on HK-2 cells in vitro

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Author:
No author available
Journal Title:
JOURNAL OF CHINESE PHYSICIAN
Issue:
6
DOI:
10.3760/cma.j.issn.1008-1372.2011.06.005
Key Word:
甲基胍/副作用;肌酸酐/副作用;肾小管/细胞学;上皮细胞/药物作用/病理学;Methylguanidine/AE;Creatinine/AE;Kidney tubules/CY;Epithelial cells/DE/PA

Abstract: Objective To investigate the effect of methylguanidine and 1-methylhydantoin on cells cytotoxicity, apoptosis in human renal tubular epithelial cell line (HK-2). Methods Human PTEC cell line HK-2 was used in this study. HK-2 was cultured and divided into 3 groups: Norma1 control group (A), methylguanidine group(B) and 1-methylhydantoin group (C). The cell inhibitory rate of HK-2 was detected by MTT method. The cytotoxicity of methylguanidine to HK-2 was determined by NAG release test. Cell apoptosis was evaluated by using Hoechst stain and FACS with Annexin-V/PI. Results The OD value and NAG concentration of creatinine, methylguanidine and 1-methylhydantoin group were compared with normal control group. OD value decreased and NAG concentration significantly increased(0.188±0.011, 0.176±0.010 vs 0.545±0.021, F=1557.74, P<0.01; 20.488±0.473, 22.225±0.565 vs 5.125±0.198, F=3848.22, P<0.01). By Hoechst stain, pycnosis and apoptotic body could be found when HK-2 was cultivated in methylguanidine 1-methylhydantoin group. In methylguanidine, 1-methylhydantoin group apoptotic HK-2 apparently increased, compared with that in control group (18.23±1.1581, 20.22±1.1433 vs 2.473±0.321, F=526.06, P<0.01). Compared with group B, the OD value in group C decreased significantly (0.176±0.010 vs 0.188±0.011,t=2.26, P<0.05), NAG concentration increased significantly (22.225±0.565 vs 20.488±0.473,t=-6.67, P<0.01), and apoptotic rate in-creased significantly (20.22±1.1433 vs 18.23±1.1581,t=-2.762, P<0.05). Conclusions 1-methylhydantoin has more powerful cytotoxic effect to renal tubular epithelial cells than that of Methylguanidine.

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