E el r2856

Serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were assessed biochemically using rat-specific ELISA kits (Cat. No. E-EL-R2856 and E-EL-R0015, respectively, Elabscience, Wuhan, China). Moreover, the C-reactive protein (CRP) was assessed using an ELISA kit from Elabscience (Cat. No. E-EL-R0506, Elabscience, Wuhan, China) following the guidelines of the manufacturer.

The serum cytokines; tumor necrosis factor-alpha (TNF-α), interferon gamma (IFN-γ) and interleukin-1 beta (IL-1β), were measured by specific ELISA kits (Elabscience, USA catalog numbers E-EL-R2856, E-EL-R0009, and E-EL-R0012, respectively) based on quantitative sandwich technique. The optical density values were read in a micro plate reader at 450 nm. The concentrations of cytokines in samples were determined by comparing the OD of the samples to the standard curve of each measured cytokine. The results were expressed as pg/mg.

The level of myeloperoxidase (MPO) was estimated in lung homogenate in freshly prepared lysis buffer, which was centrifuged for 5 min at 5000 RPM. The supernatant was collected and immediately estimated using the procedure stated by the Real gene rat MPO ELISA kit (Cat No. 3100574, Lot No.17517601575) at 435 nm (Klebanoff, 2005 (link)). The level of lipid peroxides was measured as a a thiobarbituric acid reacting substance (TBARS) and expressed as equivalent to malondialdehyde (MDA) using 1,1,3,3-tetra methoxy propane (TEP) as standard in lung homogenate (Ohkawa et al., 1979 (link)). It was expressed in terms of μ mol/mg protein. The presence of reactive substances with thiobarbituric acid (TBARS) was detected by monitoring the changes in absorbance using a microplate reader at 535 nm. TNF-α was estimated in the lung homogenate through a procedure stated by the Elabscience rat ELISA kit (E-EL-R2856).