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Objective To investigate the effect of Mysm1 on the differentiation of neural stem cells(NSCs)into astrocytes and the possible mechanism.Methods NSCs were prepared from E12.5 cortices of wild-type C57BL/6 mice,cultured in vitro and induced to differentiate into astrocytes.Immunofluorescence staining,real-time quantitative PCR and Western blot assay were used to detect the expressions of Mysm1 during the differentiation of NSCs into astrocytes in vitro.Lentivirus was used to knock down Mysm1 expressions in NSCs before real-time quantitative PCR and Western blot assay were used to detect the knockdown efficiency.Immunofluorescence staining and Western blot assay were used to compare the differentiation of NSCs into astrocytes before and after Mysm1 knockdown in vitro.Transcriptomics was adopted to detect the differential gene after knockdown of Mysm1 in NSCs in vitro.Western blot assay was used to verify the changes of proteins associated with the differential gene.Cut-Tag was used to detect the enrichment of Mysm1 in the promoter region of glial fibrillary acidic protein(GFAP)genes during the differentiation of NSCs into astrocytes in vitro.After overexpression of GFAP following knockdown of Mysm1,immunofluorescence staining and Western blot assay were used to compare the differentiation of NSCs into astrocytes before and after overexpression in vitro.Results The expression of Mysm1 was gradually increased when NSCs were induced to differentiate into astrocytes in vitro.Mysm1 knockdown inhibited the differentiation of NSCs into astrocytes in vitro.Mysm1 affected the differentiation of NSCs into astrocytes by regulating the expression of GFAP.Overexpression of GFAP after Mysm1 knockdown partially rescued the ability of NSCs to differentiate into astrocytes.Conclusion Mysm1 regulates the differentiation of NSCs into astrocytes by epigenetically controlling GFAP transcription.
RÉSUMÉ
Objective:To investigate the effects and mechanism of cannabinoid (CBD) on the anxiety- and depression-like behaviors induced by lipopolysaccharide (LPS) in mice.Methods:C57BL/6J mice were intraperitoneally injected with LPS to establish the model of neuroinflammation. CBD was injected intraperitoneally 24 h after modeling. Behavioral tests were performed to evaluate the anxiety- and depression-like behaviors in mice. CBD-pretreated BV-2 microglia cells were stimulated with LPS in vitro. The levels of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β) and CD86 in mouse cerebral cortex, hippocampus, prefrontal cortex and BV-2 cells were measured by qRT-PCR. The protein level of nuclear factor (NF-κB) in mouse brain and BV-2 cells was determined by Western blot. Results:CBD significantly increased the residence time and movement distance of LPS-treated mice in the central area in the open filed test (OFT), and reduced the immobility time in tail suspension test (TST) and force swimming test (FST). In addition, CBD alleviated the neuroinflammation and inhibited the activation of microglia in mouse brain. In vitro, CBD significantly inhibited the activation of BV-2 microglia cells. Both in vivo and in vitro experiments confirmed that CBD could inhibit NF-κB expression. Conclusions:LPS could induce the activation of BV-2 microglia cells and the expression of inflammatory factors in mouse brain accompanied with abnormal behaviors. CBD could inhibit the activation of microglia, alleviate the neuroinflammation in different regions of mouse brain and improve behavioral performance.
RÉSUMÉ
Reviewing Confucian view on justice and profit,we find that it is still of great constructive significance.The morality-lacking situation in the current drug field enlightens us to recognize the importance of ethical regulations,correctly handle the dialectic relation between justice and profit,and strive to construct a new and time-adaptive view on justice and profit.