Selection of the most effective small interfering RNA inhibiting the expression of BDNF in microglial cells and detection of its inhibitory effect / 中国药理学通报

ABSTRACT

Aim To pick out the siRNA which could most effectively inhibit the expression of brain-derived neurotrophic factor( BDNF) in microglial cells,to detect the cytotoxicity of the transfection complex,and to ob-serve the change of OX-42 expression,the microglial marker,after BDNF siRNA treatment. Methods Four siRNAs were chemically synthesized three of them were used to inhibit BDNF expression in microglial cells,the rest was fluorescence-labeled mismatch siRNA as a negative control. They were all transfected into microglial cells,respectively. BDNF mRNA was detected 24 h after transfection by Real-Time PCR and itsprotein expression was observed done by Western blot 48 h later. The Sulforhodamine B( SRB) assay was used to investigate the drug-induced cytotoxicity. Co-expres-sion pattern of BDNF and OX-42 was determined by double-labeling immunofluorescence. Results ① The BDNF siRNA1588 was the most effective siRNA,compared with the vehicle or mismatch siRNA-treated group( P