目的 观察体外培养的星形胶质细胞在炎性刺激下的激活及功能变化。 方法 体外培养 C57BL/6 小鼠皮层的星形胶质细胞，应用免疫细胞化学和Western blot等方法，测定炎性刺激下各组细胞形态学及促炎因子（iNOS、IL-6、TNF-α）和抗炎因子（ARG1、IL-10、TGF-β1）表达水平的变化情况。 结果 炎性刺激后，免疫荧光细胞染色结果显示，M1（LPS+IFN-γ）组细胞被异常激活，胞体出现肿胀变大、扭曲等形态改变，且GFAP阳性细胞数量明显增多（P<0.001 vs. control））。CCK-8细胞活力检测结果显示，LPS组、IFN-γ组及M1组的吸光度均升高，其中M1组显著升高（P<0.001 vs. control）。Western blot结果表明，M1组促炎因子（iNOS、IL-6和TNF-α）表达均明显升高（P<0.001 vs. control）；各组抗炎因子（ARG1、IL-10 和TGF-β1）的表达也显著升高（P<0.01 or 0.001 vs. control）。 结论 体外炎性刺激异常激活星形胶质细胞，形态和功能显著变化，且A1样星形胶质细胞的炎性因子表达水平升高。
Abstract
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.
关键词
星形胶质细胞 /
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脂多糖 /
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炎性刺激 /
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神经炎症
Key words
Astrocytes /
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Lipopolysaccharide /
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Inflammatory stimulation /
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Neuroinflammation
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基金
宁夏自然科学基金项目（2022AAC03153、2022AAC 03384）；国家自然科学基金（31760279）
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