I have tested the hepatoprotective effect of a natural compound in cyclophosphamide-induced rats and I want to test it in vitro using HepG2 cell line. Because my main target is oxidative stress, I want to induce oxidative stress in HepG2 cells using cyclophosphamide. I found other models like CCl4, cyclosporine and hydrogen peroxide induced oxidative stress in HepG2, but I didn't found any about cyclophosphamide. Do you have any idea?
Protein assay for heme oxygenase-1 (HO-1) induced by chemicals in HepG2 cells.
Miyamoto Y, Ohshida K, Sasago K.
Abstract
Levels of heme oxygenase-1 (HO-1), a stress response protein, were measured to examine oxidative stress induced by several chemicals in HepG2 cells with and without S9mix using an ELISA. CdCl(2), heme, and diclofenac sodium salt (diclofenac) were used as inducers of HO-1. Acetaminophen (AAP) and cyclophosphamide (CP) were used as oxidative stress inducers. Stannic mesoporphyrin (SnMP) was used as an inhibitor of HO activity. Cytotoxicity was determined, and HO-1 levels were measured in HepG2 cells exposed to chemicals other than CP with non-metabolic activation without S9mix, and to diclofenac, AAP and CP with metabolic activation with S9mix. HO-1 levels were increased by CdCl(2) (7.5 microM), heme (10, 100 microM), and stannic mesoporphyrin (SnMP) (10 microM), but were not changed by AAP, and were decreased by diclofenac. HO-1 levels were increased by diclofenac (300 microM), and CP (36 microM), but were unaffected by AAP because of low sensitivity in HepG2 cells. The induction of HO-1 expression was first observed in cultured HepG2 cells treated with CP under conditions involving metabolic activation. These results showed the measurement of HO-1 protein levels in this system is useful when assessing oxidative stress as a tool for detecting drug toxicity.
http://www.ncbi.nlm.nih.gov/pubmed/19952508
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