Advanced oxidation protein products induce M1 activation of BV2 cells: an experimental study

Abstract

Objective To investigate whether advance oxidation protein products (AOPPs) would induce M1 activation of BV2 cells. Methods AOPPs were prepared by incubation of mouse serum albumin (MSA) with sodium hypochlorite solution. Methyl thiazol tetrazolium (MTT)assay was used to determine the effects of different concentration of AOPPs on BV2 cells activity by incubating the cells for 24 h for selecting optimum working concentration of AOPPs. BV2 cells were treated based on concentration effect. Intracellular reactive oxygen species (ROS) production was determined with dichlorofluorescein diacetate (DCFH-DA) and Confocal laser scanning microscope system. And the protein expression of nicotinamide adenine dinucleotide (NADPH) oxidase (NOX)4 (NOX4), cluster of differentiation molecule 11b (CD11b) and tumor necrosis factor (TNF-α) were examined by Western blotting. Results Viability of BV2 cells treated with 400 μg/ml and 800 μg/ml AOPPs was significantly lowered [(50.052±2.163)%, (36.505±1.589)%, P=0.000]. AOPPs, in a concentration-dependent manner, increased the expression of NOX4, CD11b and TNF-α (NOX4: PLPS=0.001, P50 μg/ml=0.000, P100 μg/ml=0.004, P200 μg/ml=0.000; CD11b: P=0.000; TNF-α: P=0.000); prolonged exposure to AOPPs and increase of AOPPs concentration both led to an increase of intracellular ROS production (P=0.000). Conclusion AOPPs may induce M1 activation of BV2 cells through producing ROS. Key words: Advance oxidation protein products; BV2 cells; Reactive oxygen species; M1 activation