I followed a protocol that I saw in many papers to promote liver injury by ethanol challenge in C57bl/6 mice but my results are completely different.

The levels of ALT agree with the papers but MDA and GSH levels are opposite.

The protocol is:

3 doses of ethanol 5g/Kg 32 or 50% (12h-12h) and euthanasia after 4h of the last dose. At this way MDA should increase and GSH should decrease, but my results showed the opposite.

So, I changed the dose (56%; 6g/Kg) and the time until the euthanasia (12h) but the values still continue at the same way.

Techniques used: ALT and GSH by spectrophotomer; MDA-DNPH by HPLC

Could Someone who works with oxidative stress help me?