Brucella outer membrane protein Omp25 induces microglial cells in vitro to secrete inflammatory cytokines and inhibit apoptosis.

Omp25 protein, an outer membrane protein of Brucella, can cause damage to the central nervous system. As one type of macrophage, microglial cells play a role in immune surveillance and immune protection in the central nervous system; therefore, they are major targets of bacterial attack. The present study examined BV2 mouse microglial cells that were stimulated with different concentrations of Omp25 recombinant protein, and the secretion of inflammatory cytokines by the BV2 cells as well as their level of apoptosis were observed. The objective of the study was to preliminarily illustrate the possible mechanism that Omp25 uses to damage the central nervous system. Mouse BV2 microglial cells were incubated with different concentrations of Omp25 for 24 h, and an enzyme-linked immunosorbent assay (ELISA) was used to detect the secretion of the inflammatory cytokines interleukin (IL)-6, tumour necrosis factor (TNF)-α and HMGB1 (high mobility group box-1 protein); reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of TLR4 (Toll-like receptor 4) mRNA; Annexin V-fluorescein isothiocyanate (FITC) double staining was used to detect apoptosis in the BV2 cells. After the BV2 cells were stimulated with different concentrations of Omp25, the levels of IL-6, TNF-α and HMGB1 was increased, and the difference was statistically significant compared with the control group (P<0.05). The secretion of TNF-α and HMGB1 showed a trend toward an initial increase followed by a decrease. The expression level of TLR4 mRNA was increased. Omp25 protein can inhibit apoptosis in BV2 cells. The outer membrane protein Omp25 of Brucella promotes microglial cells to secrete inflammatory cytokines and inhibit apoptosis. TLR4 may be involved in the immune response of the central nervous system to Brucella infection.

[Effect of polymeric aluminum-iron on EPS and bio-flocculation in A2/O system].

Polymeric aluminum-iron (PAFC) was added at the end of aeration tank to enhance phosphorus removal, so that the phosphorus concentration in the effluent could meet the calss A standard in municipal sewage treatment plant pollutant discharge standard (GB 18918-2002). The characteristics of extracellular polymer substances (EPS) and bio-flocculation for the activated sludge in the A2/O system were analyzed in the experiment. The results showed that, the gross of EPS varied little with the increase in PAFC dosage, while, the ratio of albumen to polysaccharide declined from 3.30 to 2.30. When the PAFC dosage increased, the concentration of Al3+ in EPS increased during the whole anaerobic-anoxic-aerobic cycle. The flocs of activated sludge became larger after PAFC addition, Zeta potential of the effluent dropped significantly from - 15.83 mV to -21.20 mV and sludge yield increased. Therefore, bio-flocculation of the activated sludge in the A2/O system improved when a proper amount of PAFC was added, subsequently improve the water quality of the effluent.