Chinese Journal of Clinical Anatomy ›› 2024, Vol. 42 ›› Issue (1): 42-46.doi: 10.13418/j.issn.1001-165x.2024.1.08

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Activation and functional changes of astrocytes cultured in vitro under inflammatory stimulation

Cheng Yingying1,2, Wu Jianjun1, Cai Haiyan 3, Jiao Xunwen1, Ma Jiangbo1, Ding Yinxiu1*   

  1. 1. Department of Human Anatomy, Basic Medical College, Ningxia Medical University, Yinchuan 750006, China; 2.Department of Neurosurgery, The Second Affiliated Hospital of Xi’an Medical University, Xi'an 710038, China; 3. Department of Neurology, the People’s Hospital, Ningxia Hui Autonomous Region, Yinchuan 750006, China 
  • Received:2023-07-04 Online:2024-01-25 Published:2024-01-30

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.  

Key words: Astrocytes,  ,  , Lipopolysaccharide,  ,  , Inflammatory stimulation,  ,  , Neuroinflammation

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