体外炎性诱导星形胶质细胞激活及功能分析

doi:10.13418/j.issn.1001-165x.2024.1.08

中国临床解剖学杂志 ›› 2024, Vol. 42 ›› Issue (1): 42-46.doi: 10.13418/j.issn.1001-165x.2024.1.08

体外炎性诱导星形胶质细胞激活及功能分析

程莹莹^{1, 2}，武建军¹，蔡海燕³，焦旭文¹，马江波¹，丁银秀¹*

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

收稿日期:2023-07-04 出版日期:2024-01-25 发布日期:2024-01-30
通讯作者: 丁银秀，博士，教授，E-mail: yinxiuding@163.com
作者简介:程莹莹（1992-），女，陕西西安人，硕士研究生，E-mail: 594028773@qq.com
基金资助:
宁夏自然科学基金项目（2022AAC03153、2022AAC 03384）；国家自然科学基金（31760279）

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

Cheng Yingying^1,2, Wu Jianjun¹, Cai Haiyan ³, Jiao Xunwen¹, Ma Jiangbo¹, Ding Yinxiu¹*

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

摘要/Abstract

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。

关键词: 星形胶质细胞, , , 脂多糖, , , 炎性刺激, , , 神经炎症

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

中图分类号:

程莹莹, 武建军, 蔡海燕, 焦旭文, 马江波, 丁银秀. 体外炎性诱导星形胶质细胞激活及功能分析[J]. 中国临床解剖学杂志, 2024, 42(1): 42-46.

Cheng Yingying, Wu Jianjun, Cai Haiyan , Jiao Xunwen, Ma Jiangbo, Ding Yinxiu. Activation and functional changes of astrocytes cultured in vitro under inflammatory stimulation[J]. Chinese Journal of Clinical Anatomy, 2024, 42(1): 42-46.

参考文献 16

[1]	Grigoriadis N, van Pesch V, Paradig MSG. A basic overview of multiple sclerosis immunopathology[J]. Eur J Neurol, 2015, 22(Suppl2): 3-13. DOI: 10.1111/ene.12798.
[2]	Sofroniew MV. Multiple roles for astrocytes as effectors of cytokines and inflammatory mediators[J]. Neuroscientist, 2014, 20(2): 160-172. DOI: 10.1177/1073858413504466.
[3]	Niranjan R. The role of inflammatory and oxidative stress mechanisms in the pathogenesis of Parkinson’s disease: focus on astrocytes[J]. Mol Neurobiol, 2014, 49(1): 28-38. DOI: 10.1007/s12035-013-8483-x.
[4]	Koob AO. Astrogenesis versus astrogliosis[J]. Neural Regen Res, 2017, 12(2): 203-204. DOI: 10.4103/1673-5374.200798.
[5]	Masuda T, Sankowski R, Prinz M, et al. Microglia heterogeneity in the single-cell era[J]. Cell Reports, 2020, 30(5): 1271-1281. DOI: 10.1016/j.celrep.2020.01.010.
[6]	Glass CK, Saijo K, Gage FH, et al. Mechanisms underlying inflammation in neurodegeneration[J]. Cell, 2010, 140(6): 918-934. DOI: 10.1016/j.cell.2010.02.016.
[7]	Morita M, Ikeshima-Kataoka H, Noda M, et al. Metabolic Plasticity of Astrocytes and Aging of the Brain[J]. Int J Mol Sci, 2019, 20(4): 941. DOI: 10.3390/ijms20040941.
[8]	Kery R, Chen APF, Kirschen GW. Genetic targeting of astrocytes to combat neurodegenerative disease[J]. Neural Regen Res, 2020, 15(2): 199-211. DOI: 10.4103/1673-5374.265541.
[9]	Ajmone-Cat MA, Mancini M, Minghetti L, et al. Microglial polarization and plasticity: evidence from organotypic hippocampal slice cultures[J]. Glia. 2013, 61(10): 1698-1711. DOI: 10.1002/glia.22550.
[10]	Zhang K, Zheng J, Wang J, et al. Polarized macrophages have distinct roles in the differentiation and migration of embryonic spinal-cord-derived neural stem cells after grafting to injured sites of spinal cord[J]. Mol Ther. 2015, 23(6):1077-1091. DOI: 10.1038/mt.2015.46.
[11]	Sochocka M, Diniz BS, Leszek J. Inflammatory response in the CNS: friend or foe[J]? Mol. Neurobiol, 2017, 54(10): 8071-8089. DOI: 10.1007/s12035-016-0297-1.
[12]	Sofroniew MV. Astrocyte barriers to neurotoxic inflammation[J]. Nat Rev Neurosci. 2015, 16(5):249-63.DOI: 10.1038/nrn3898.
[13]	Cekanaviciute E, Dietrich HK, Wai KM, et al. Astrocytic TGF-β signaling limits inflammation and reduces neuronal damage during central nervous system Toxoplasma infection[J]. J Immunol. 2014,193(1):139-49.DOI: 10.4049/jimmunol. 1303284.
[14]	Cynthia JM Kane, Paul D Drew. Neuroinflammatory contribution of microglia and astrocytes in fetal alcohol spectrum disorders[J]. J Neurosci Res, 2021,99(8):1973-1985. DOI: 10.1002/jnr.24735.
[15]	Sidoryk-Wegrzynowicz M, Strużyńska L. Dysfunctional glia: contributors to neurodegenerative disorders[J]. Neural Regen Res, 2021, 16(2): 218-222. DOI: 10.4103/1673-5374.290877.
[16]	Li KY, Li JT, Qin S, et al. Reactive Astrocytes in Neurodegenerative Diseases[J]. Aging and Disease, 2019, 10(3): 664-675. DOI: 10.14336/AD.2018.0720.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献 16

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	江剑宏, 段仁鹏, 李晓锋. 国人胆道三维重建解剖变异应用研究[J]. 中国临床解剖学杂志, 2024, 42(1): 1-4.
[2]	郭红志, 王瑜, 张加琪, 梁海彬, 马梓玮, 冯伟, 吴忧, 司子燚. 血管铸型结合CT三维模型评估先兆子痫胎盘血管结构[J]. 中国临床解剖学杂志, 2024, 42(1): 5-10.
[3]	李佳伟, 张静, 李灿然, 兰文杰, 籍庆余, 郭志勇, 张云凤, 刘启, 陈清威, 李筱贺. 蒙古族人群股骨近端解剖参数X线测量[J]. 中国临床解剖学杂志, 2024, 42(1): 11-16.
[4]	李琨, 张燕, 郭冉, 郝咪咪, 吴轩宇, 徐艺芳, 王超群, 马文童, 张灵淇, 杨宏宇, 李志军, 张少杰, 王星. 儿童及青少年枕颈角与后枕颈角数字化测量及其临床意义[J]. 中国临床解剖学杂志, 2024, 42(1): 17-20.
[5]	李文, 杨思艺, 黄蕾, 卿霁雯, 蒋松涛, 张磊. 基于MRI探讨Lisfranc韧带的形态学特点及临床意义[J]. 中国临床解剖学杂志, 2024, 42(1): 21-25.
[6]	李凡凡, 徐阳, 王晓旭. 紫草素调节Nrf2/HO-1信号通路对实验性大鼠肉芽肿性小叶性乳腺炎的治疗作用研究[J]. 中国临床解剖学杂志, 2024, 42(1): 26-32.
[7]	王兴航, 丁佳媛, 李放, 包翠芬, 阎丽菁. 人参皂苷Rg1通过抑制NLRP3炎症小体途径减轻氧糖剥夺/复供后小胶质细胞炎症反应[J]. 中国临床解剖学杂志, 2024, 42(1): 33-41.
[8]	祁志, 杨力, 贾秋叶, 陈浩伦, 段兆达, 吴春云. 依达拉奉经MAPKs通路对脂多糖激活的小胶质细胞Sirt3表达调控[J]. 中国临床解剖学杂志, 2024, 42(1): 47-53.
[9]	张晶晶, 王洪新. 黄芩苷通过PDGF/P38 MAPK信号通路对肺动脉高压大鼠的保护作用[J]. 中国临床解剖学杂志, 2024, 42(1): 54-58.
[10]	乐钦, 陈荣丽, 熊婧熙, 王婷怡, 蔡志恒, 易秋实, 曾心怡. 白芨多糖对大鼠跨区皮瓣choke区的影响[J]. 中国临床解剖学杂志, 2024, 42(1): 59-64.
[11]	李笑予, 张磊, 付磊, 李东波, 汪国友. 三步接骨法治疗Sanders Ⅱ型跟骨骨折的有限元研究[J]. 中国临床解剖学杂志, 2024, 42(1): 65-70.
[12]	吴研飞, 马剑雄, 卢斌, 王颖, 柏豪豪, 靳洪震, 马信龙. CT后处理技术重建CT值与终板抗压强度的相关性研究[J]. 中国临床解剖学杂志, 2024, 42(1): 71-76.
[13]	庄万强, 唐毅, 骆勇刚, 张辉. 关节镜联合胫骨高位截骨术对髌骨位置及髌股关节功能影响的早中期回顾性研究[J]. 中国临床解剖学杂志, 2024, 42(1): 77-82.
[14]	罗鹰, 窦伟誉, 吴小杭, 陈璟昆, 彭昌贵, 潘剑英 . 微创海马钢板内固定治疗跟骨骨折并发症的影响因素[J]. 中国临床解剖学杂志, 2024, 42(1): 83-88.
[15]	陈俊臣, 曹文英, 罗燕君, 何婉仪, 朱真真, 彭志强. 抽吸取栓术与抽吸结合支架取栓术治疗脑前循环栓塞疗效比较[J]. 中国临床解剖学杂志, 2024, 42(1): 89-93.