体外炎性诱导星形胶质细胞激活及功能分析  

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

作　　者：程莹莹 武建军 蔡海燕[3] 焦旭文[1] 马江波[1] 丁银秀[1] Cheng Yingying;Wu Jianjun;Cai Haiyan;Jiao Xunwen;Ma Jiangbo;Ding Yinxiu(Department of Human Anatomy,Basic Medical College,Ningxia Medical University,Yinchuan 750006,China;Department of Neurosurgery,The Second Affiliated Hospital of Xi’an Medical University,Xi'an 710038,China;Department of Neurology,the People’s Hospital,Ningxia Hui Autonomous Region,Yinchuan 750006,China)

机构地区：[1]宁夏医科大学基础医学院人体解剖学系,宁夏银川750006 [2]西安医学院第二附属医院神经外科,陕西西安710038 [3]宁夏回族自治区人民医院神经内科,宁夏银川751006

出　　处：《中国临床解剖学杂志》2024年第1期42-46,共5页Chinese Journal of Clinical Anatomy

基　　金：宁夏自然科学基金项目(2022AAC03153、2022AAC03384);国家自然科学基金(31760279)。

摘　　要：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。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-γgroup 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-αin M1 group were markedly increased after LPS and IFN-γcombined 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 脂多糖 炎性刺激 神经炎症 

分 类 号：R3[医药卫生—基础医学]