Molecular Mechanism Of Resveratrol Improving Learning And Memory Impairment In Manganese-induced Mice By Up-regulating PGC-1α Signaling

ObjectiveTo study the effects of RSV on learning and memory impairment in mice induced by manganese,and to explore its possible molecular regulatory mechanism.To provide experimental and theoretical basis for the treatment of manganese poisoning and other neurodegenerative diseases.MethodsThis study is divided into in vivo experiment and in vitro experiment.Forty adults Kunming mice,half male and half female,were randomly divided into 4groups: control group,Mn group,RSV group and RSV +Mn group.Morris water maze test was used to analyze the learning and memory ability of mice.Manganese accumulation in hippocampus was detected by flame atomic absorption spectrometry.The apoptosis of mice hippocampus nerve cells was analysed by Nith staining.The m RNA expression levels of M1/M2 microglia markers,Pgc-1α and Bdnf were detected by q PCR.The expression levels of inflammatory cytokines in hippocampus nerve cells of mice were analyzed by ELISA.The protein expression levels of PGC-1α,p62,ULK1,LC3,p-NF-κB and BDNF were detected by Western blotting.The acetylation levels of STAT6 and ULK1 in hippocampus nerve cells were analyzed by immunoprecipitation.BV2 cells were transfected with PGC-1α lentivirus in vitro.CCK8 assay was used to detect cell viability.The q PCR was used to detect the m RNA expression of M1/M2 microglia markers.The expression levels of PGC-1α and autophagy-related proteins were detected by Western blot.A tandem fluorescent m Cherry-GFP-LC3 B reporter was used to monitor autophagic flux.ResultsIn vivo experiments showed that in the Morris water maze experiment,compared with the Mn group,the escape latencies and mean distances of mice in RSV+Mn group were significantly reduced,while the crossing times and time in target quadrant were significantly increased.RSV increased the number of hippocampal surviving nerve cells and alleviated the damage of hippocampal neuron.The protein expression levels of ULK1 and LC3-II in RSV+Mn group were significantly higher than those in Mn group,while the protein expression levels of p62 were significantly decreased.The protein and m RNA expression levels of PGC-1α and BDNF in RSV+Mn group were significantly increased compared with those in Mn group.RSV intervention decreased the acetylation level of STAT6 protein,decreased the m RNA expression level of M1 microglia and increased the m RNA expression level of M2 microglia.In vitro experiments showed that,compared with RSV+Mn group,the expression levels in BV2 cells of PGC-1α,ULK1 and LC3-II protein were significantly decreased and the expression levels of p62 protein were significantly increased after transfected with LV-PGC-1α sh RNA.By silencing PGC-1α,the RSV increase in m RNA expression level of M1 microglia and decrease in m RNA expression level of M2 microglia were blocked.Results of detection of autophagic flux showed that yellow signal dots significantly increased,while red signal dots significantly decreased after down regulation of PGC-1α compared with RSV+Mn group,indicating that RSV improved mangano-induced autophagy flow blocking effect disappeared.Conclusion1.Resveratrol can improve manganese-induced hippocampal learning and memory impairment in mice.2.Resveratrol can inhibit the NF-κB signal and the acetylation of STAT6 by up-regulating PGC-1α,inhibit the M1 polarization of microglia cells,promote M2 polarization,inhibit neuroinflammation,and improve learning and memory disorders.3.Resveratrol can promote the expression of ULK1 by up-regulating PGC-1α,improve the autophagy disorder induced by manganese,and alleviate the impairment of learning and memory.