Eeffect of HMGB1 on expression of NF-κB in BV-2 cells stimulated with Aβ25-35 / 中国病理生理杂志

ABSTRACT

AIM:To investigate the effect of high mobility group box-1 protein (HMGB1) on the expression of nuclear factor-κB ( NF-κB) in BV-2 cells stimulated with amyloid β-protein ( Aβ) 25-35 .METHODS:Cultured BV-2 cells in logarithmic growth phase were divided into 4 groupsnormal cell group ( without any treatment ) , model group ( treated with Aβ25-35 at 40 μmol/L) , RNA interference ( RNAi) group ( conducted with HMGB1-siRNA followed by Aβ25-35 stimula-tion) and solvent control group (treated with 0.1% DMSO).After treatment with Aβ25-35 for 24 h, the protein levels of HMGB1 and NF-κB in BV-2 cells were determined by Western blot .RESULTS:Aβ25-35 at 40μmol/L was used to stimu-late BV-2 cells.The GFP fluorescence-tagged HMGB1-siRNA (30 nmol/L) was used to transfect BV-2 cells and its trans-fection efficiency was about 80%~90%.The results of Western blot showed that the protein level of HMGB 1 was signifi-cantly decreased after the interference of siRNA fragment (P<0.05).The protein levels of HMGB1 and nucleic NF-κB p65 were dramatically increased in BV-2 cells stimulated with Aβ25-35(P<0.05).After RNA interference with HMGB1, the expression of HMGB1 and nucleic NF-κB p65 were significantly decreased in BV-2 cells stimulated with Aβ25-35 ( P<0. 05).CONCLUSION:RNA interference with HMGB1 reduces the expression of nucleic NF-κB in BV-2 cells stimulated with Aβ25-35 .