*4.7. Biochemical Analysis*

Mice were sacrificed by cervical vertebrae removal. The kidneys were carefully removed, washed with 0.9% NaCl, frozen in liquid nitrogen, and stored at −80 ◦C for subsequent physical and chemical analysis. Kidney specimens were homogenized with tissue homogenizer in phosphate buffer pH 7.4, and the supernatant was collected centrifugally (3000 rpm, 10 min). The contents of MDA, SOD, GSH-Px, and T-AOC were measured according to the kit instructions, and the renal antioxidant enzyme activity was evaluated. The contents of protein carbonyl and 8-OHdG were determined to evaluate the oxidative damage level of protein and DNA. The levels of TNF-α and IL-6 in the kidney were determined by ELISA, and the levels of inflammatory factors were evaluated.

### *4.8. Hematoxylin and Eosin (H&E) Staining*

Kidney specimens were fixed in 10% formaldehyde normal saline for 5–7 days, washed with clean water for 10 min, and dehydrated with alcohol classification (75%, 85%, 95%, 100%) in accordance with conventional sequence, and then they were treated with xylene to make them transparent. The specimen was then embedded in paraffin. The specimens were cut into 5 mm slices by a slicer. The tissues were stained with hematoxylin and observed under an optical microscope.
