Rat reactive oxygenspecies cluster kit

Parotid gland tissues were rapidly removed, thoroughly homogenized and
centrifuged at 5000 rpm/min for 10 minutes. The supernatant was assayed in
accordance with the manufacturer’s instructions for the Rat Reactive Oxygen
Species Cluster Kit (Mlbio, Shanghai, China). The optical density value for each
specimen was determined by a microplate reader at 450 nm (FilterMax F3,
Molecular Devices Corporation, San Francisco, CA). Samples for total antioxidant
capacity (T-AOC) detection were homogenized and centrifuged at 10 000 rpm/min
for 5 minutes. The supernatant was assayed in accordance with the manufacturer’s
instructions for the T-AOC Kit (Solarbio). The optical density value for each
specimen was determined by a microplate reader at 593 nm (MD, FilterMax F3).
According to the manufacturer’s instructions, the levels of tumor necrosis
factor-alpha (TNF-α), interleukin (IL)-6, and IL-2 in the rat serum were
determined using a TNF-α enzyme-linked immunosorbent assay (ELISA) kit (R&D,
Sao paulo, MN), an IL-6 ELISA kit (R&D), and an IL-2 ELISA kit (R&D),
respectively.

Lung tissues were homogenized and centrifuged at 5000 rpm/min for 10 min. The supernatant was assayed in accordance with the manufacturer’s instructions for the Rat Reactive Oxygen Species Cluster Kit (Mlbio, Shanghai, China). The optical density value for each specimen was determined by a microplate reader at 450 nm (FilterMax F3, Molecular Devices Corporation, San Fran-cisco, CA).