*4.4. Experimental Model*

The animals were divided into four groups of six rats in each group. Animals in Group I served as normal control and were treated with normal saline. Animals in Group II served as disease control and IV were treated with LPS (10 mg/kg, i.p). Group III animals were treated with ZIN 150 mg/kg p.o 2 h before LPS challenge. Group IV served as positive control and were treated with ZIN 150 mg/kg p.o.

#### *4.5. Determination of Biochemical, Oxidative Stress and Inflammatory Markers*

Animals were euthanized with isoflurane after six hours of LPS challenge and blood samples were collected in sterilized EDTA tubes. Different organs like liver, lung, kidney, and brain tissues were harvested. The plasma samples were stored at −80 ◦C for biochemical, oxidative, and inflammatory marker analyses. Biochemical parameters like ALT, ALP, AST, BUN, Cr, Urea, LDH, Albumin, bilirubin, total protein, and CKMB were assessed. Oxidative stress markers MDA, GSH, MPO, and (DNA damage marker) 8-OHdGlevels were measure by ELISA technique as per the manufacturer details. Levels of nitric oxide (NO) were measured in plasma by Griess reaction method. The color intensity was recorded at 540 nm using ELISA plate reader. Inflammatory markers like TNF-<sup>α</sup>, IL-1β, IL-1<sup>α</sup>, IL-2, IL-6, and IL-10 were quantified using ELISA kits as per their manufacturer protocol. The plasma procalcitonin, a septic biomarker was determined using ELISA kits obtained according to the manufacturer's instructions.
