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Title: Metal-induced oxidative damage in cultured hepatocytes and hepatic lysosomal fraction: Beneficial effect of a curcumin/absinthium compound
Authors: R. Barreto
S. Kawakita
J. Tsuchiya
E. Minelli
K. Pavasuthipaisit
A. Helmy
F. Marotta
Biokenkyujo Research Laboratory
Universita degli Studi di Milano
Mahidol University
Minufiya University
S. Guiseppe Hospital
Keywords: Medicine
Issue Date: 19-Apr-2005
Citation: Chinese Journal of Digestive Diseases. Vol.6, No.1 (2005), 31-36
Abstract: Objective: Metals undergo redox cycling and there is increasing evidence of free radical generation and oxidative injury in the pathogenesis of liver injury and fibrosis in metal storage diseases. The aim of the present study was to test a natural hepatoprotective compound in metal-induced liver injury. Methods: Hepatocytes were isolated from Wistar rats by collagenase perfusion method and cultured as such and also with α-linolenic acid (LNA)-bovine serum albumin (BSA). Hepatocytes were then cultured with a graded dilution of PN-M001 (100 μg/mL and 200 μg/mL), which is a curcuma/absinthium-containing compound, or sylibin (100 μg/mL) dissolved in dimethyl sulfoxide for 10 min before the addition of metallic salts (iron, copper and vanadium). Lysosomal fractions were prepared for lysosome fragility tests in which β-galactosidase activity and lactate dehydrogenase (LDH) leakage were measured, as well as oxidative damage tests in the presence of hydrophilic and lipophilic free radical generators. Quenching activity by 1,1-diphenyl-2-picrylhydrazyl (DPPH) was also assessed. Results: Malonildialdehyde accumulation in the medium showed a direct time-course increase with incubation time. Both PN-M001 and sylibin showed a significant protective effect against all challenge metal ions, as expressed by the half inhibition concentration (IC50) against lipid peroxidation. However, on a molar ratio, sylibin seemed to be more effective than PN-M001 in Fe-induced peroxidative damage (P < 0.05). Both test compounds, irrespective of the concentration, significantly reduced the LDH and β-galactosidase concentration in the lysosomal fractions. As compared with untreated lysosomal fractions challenged with the two peroxide radicals generators, either PN-M001 or sylibin exerted significant protection However, PN-M001 was significantly better than sylibin in suppressing acid phosphatase enzyme activity. Both compounds showed comparable and significant DPPH radical-scavenging activity. Conclusion: These data support the potential clinical application of curcumin-containing compounds.
ISSN: 14439611
Appears in Collections:Scopus 2001-2005

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