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 Al-like inflammatory factors in astrocytes are increased.
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