Objective To observe the activation and functional changes of astrocytes cultured in vitro under inflammatory stimulation. Methods Astrocytes from the cortex of C57BL/6 mice were isolated and cultured in vitro, the changes of astrocyte morphology were observed by immunocytochemistry and the expression levels of proinflammatory factors (iNOS, IL-6, TNF-α) and anti-inflammatory factors (ARG1, IL-10, TGF-β1) were determined by Western blot protein analysis. Results After inflammatory stimulation, immunofluorescence cell staining showed that astrocytes in M1（LPS+IFN-γ） group were abnormally activated, cell body swelling, enlargement, distortion and other morphological changes were also obvious, and the number of GFAP positive astrocytes in M1 group were evidently increased (P<0.001 vs. control). CCK-8 test showed that compared with the control group, the absorbance values of LPS group, IFN-γ group and M1 group were increased, especially the absorbance values of M1 group was significantly increased (P<0.001 vs. control). Western blot results showed that the expression levels of iNOS, IL-6 and TNF-α in M1 group were markedly increased after LPS and IFN-γ combined stimulation (P<0.001 vs. control). The expression levels of anti- inflammatory factors (ARG1, IL-10 and TGF-β1) in each group were also significantly upregulated (P<0.01 or 0.001 vs. control). Conclusions Under inflammatory stimulation, the cultured-astrocytes in vitro can be abnormally activated, its morphological and functional changes are distinct, and the expression level of A1-like inflammatory factors in astrocytes are increased.
Key words
Astrocytes /
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Lipopolysaccharide /
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Inflammatory stimulation /
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Neuroinflammation
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