*4.11. Immunohistochemistry Analysis*

MIP-2 and COX-2 expressions in hepatic tissue were evaluated by immunohistochemistry staining as previously described [96]. The prepared hepatic sections were recovered, and the endogenous peroxidase in tissues was inactivated with 0.1% hydrogen peroxide containing methanol for 15 min. Then, the sections were incubated with a rabbit polyclonal MIP-2 and COX-2 antibody at 4 ◦C overnight. Subsequently, the sections were washed with PBS and incubated with rabbit anti-mouse (1:1000) secondary antibody at

room temperature for 30 min. The sections were rinsed with PBS 3 times and stained with diaminobenzidine (DAB). Additionally, they were evaluated under an optical microscope (Olympus Optical Co., Ltd., Tokyo, Japan).
