RESUMO
Aim To investigate the effeets of prolifera¬tion and autophagy of BV2 eells in OGD/R models when the 18 ku transloeator protein( TSPO) was inhibi¬ted.Methods BV2 microglia were eultured in vitro and the model established by oxygen-glueose depriva- tion/reperfusion( OGD/R) , the eells were divided into eontrol group and OGD/R group, OGD/R + small hair¬pin RNA negative eontrol group ( OGD/R + NCshR- NA) , OGD/R + TSPO small hairpin RNA group (OGD/R + TSPOshRNA ).The expression of TSPO mRNA and TSPO protein were deteeted by qRT-PCR and Western blot, respectively.In order to study the effeet of TSPO on BV2 microglial eells in OGD/R inju¬ry and autophagy, the cell viability was tested by CCK- 8 assey, the cytotoxicity was deteeted by reactive oxy¬gen speeies ( ROS) , autophagy-related mRNA ( p62 mRNA, LC3B mRNA, Beolin-1 mRNA) expressions were detected by qRT-PCR, and the expression levels of autophagy -related proteins ( p62 , LC3 II /LC3 1 , Beclin-1 ) were detected by Western blot in each group.Result The expression of TSPO mRNA and protein increased significantly in OGD/R group while compared to control group, the cell death and cytotox¬icity increased significantly, the expression levels of LC3B mRNA and Beclin-1 mRNA increased, while the p62 mRNA decreased significantly, the levels of LC3 II/LC3 1 and Beclin-1 protein increased, the expres¬sion of p62 protein decreased significantly in OGD/R group, and the autophagy was activated; compared with OGD/R group, the different levels of cell viabili¬ty, cytotoxicity and autophagy in OGD/R + NCshRNA group were not statistically significant.But the survival rate of cells in OGD/R + TSPOshRNA group signifi¬cantly increased, the levels of cytotoxicity and autoph¬agy were significantly reduced.Conclusions The in¬hibition of TSPO has a significant protective effect on OGD/R injury model in BV2 microglial cells, which may be related to the inhibition of autophagy.
RESUMO
OBJECTIVE: To observe the influence of excessive fluoride on the levels of osteocalcin and testosterone in the testis of the male mouse. METHODS: Twenty-four C57BL/6J male mice were equally randomized into a normal control and a fluorosis model group, the former fed on distilled water while the latter on a solution of sodium fluoride (100 mg/L) in distilled water, both for 12 weeks. Then, the level of osteocalcin in the testis tissue was measured with the immunohistochemical streptavidin-peroxidase (SP) method and those of osteocalcin and testosterone in the serum determined by ELISA. RESULTS: After 12 weeks of fluoride intervention, the level of serum osteocalcin was significantly higher in the fluorosis models than in the normal controls (ï¼»68.05 ± 5.32ï¼½ vs ï¼»47.50 ± 5.73ï¼½ pg/mL, F = 11.901, P = 0.008), while that of testosterone markedly lower in the former than the latter group (ï¼»8.07 ± 1.35ï¼½ vs ï¼»12.94 ± 3.09ï¼½ ng/mL, F = 2.313, P = 0.006). The results of immunohistochemical SP showed the expression of osteocalcin in the cell membrane and cytoplasm of the fluorosis models, which was evidently higher than in the normal controls. CONCLUSIONS: Twelve-week intake of 100 mg/L fluoride solution can decrease the level of testosterone and increase the expression of osteocalcin in the testis of the male mouse.
