Objective To investigate the effect of fluorosis on microglia BV2 and N2a cells,and on the damages and autopany of cerebral cortical neurons of mice.Methods Eight-week-old clean-grademale mice of C57BL/6J wild-type were randomly divided intothe control group(fluoridecontent in drinking water was＜0.5 mg/L),the low-fluoride group(fluoride in drinking water was 10.0 mg/L)andthehigh-fluoride group(fluoride in drinking water was 50.0 mg/L),All the mice(10 cases in each group)were fed for 12 weeks.BV2 and N2a cells were divided into the control group,thelow-fluoride group(medium fluoride 1.0 mmol/L)and the high-fluoride group(medium fluoride 1.5 mmol/L)after normal culture.CCK8 was used to detect thechanges in proliferative viability of BV2 and N2a cells under different concentrations of fluorine treatment.Fluoride ion-selective electrode method was used to detect the changes offluoride contentsin urine,blood and bone of the modeled mice.Protein immunoblotting was used to detect the changes of the expression of ion-calcium-binding bridging molecule-1(Iba-1),which served as a marker of microglia activation,in the cortex of the mouse.Western blot test(WB)was used to detect the expression levels of autophagy-related microtubule-associated protein 1 light chain 3(LC3),and autophagy receptor protein P62(SQSTM1/P62)in N2a cells,BV2 cells and mouse cerebral cortex,and further to detectmitochondrial dynamics and autophagy fusion and division-associated proteins Mitochondrial Fusion Protein 1(Mfn1),Mitochondrial Fusion Protein 2(Mfn2),and Dynamic related protein 1(Drp1),tension protein homologue-induced putative kinase 1(Pink1),and E3 ubiquitin ligase(E3 ubiquitin-protein ligase,Parkin)expression levels.Results CCK8 showed that the value-added viability of BV2 and N2a cells treated with 1.0 mmol/L and 1.5 mmol/L NaF decreasedsignificantly(P＜0.01,P＜0.0001);the incidence of dental fluorosis in mice in the low-fluoride group was 60％(6/10),and that in the high-fluoride group was 80％(8/10),whilethe fluoride contentinurine,blood,and bone of the high-fluoride group significantly increased compared with the control group(all P＜0.001);protein immunoblotting was used to detect the changes ofthe expression of Iba-1 in BV2 and mouse cerebral cortex.LC3 and P62 in BV2,N2a cells and mouse cerebral cortex.Mfn1,Mfn2,Drp1,Pink1,and Parkin expression level changes in the expression level.Conclusion Fluorosis leads to mitochondrial damage,disrupt the balance of mitochondrial fusion and division and cause cellular autophagy,which provides a new way of thinking to elucidate the interrelationship between neurological damage and cellular autophagy in fluorosis.
fluorosisautophagymitochondrial fusion and splitting proteinsmitochondrial autophagymicroglia cellmice
维普
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