[1]陈思思　朱馨艺　邓　钢　王　龙.lncRNA LINC01089对氧糖剥夺/复氧损伤后小胶质细胞极化的影响[J].中国临床神经外科杂志,2022,27(03):186-192.[doi:10.13798/j.issn.1009-153X.2022.03.012]
　CHEN Si-si,ZHU Xin-yi,DENG Gang,et al.Effect of lncRNA LINC01089 on microglia polarization after oxygen-glucose deprivation/reoxygenation injury[J].,2022,27(03):186-192.[doi:10.13798/j.issn.1009-153X.2022.03.012]
点击复制

lncRNA LINC01089对氧糖剥夺/复氧损伤后小胶质细胞极化的影响()
分享到：

《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
27
期数:
2022年03期
页码:
186-192
栏目:
实验研究
出版日期:
2022-03-31

文章信息/Info

Title:
Effect of lncRNA LINC01089 on microglia polarization after oxygen-glucose deprivation/reoxygenation injury
文章编号:
1009-153X（2022）03-0186-07
作者:
陈思思　朱馨艺　邓　钢　王　龙
430060 武汉，武汉大学人民医院神经外科（陈思思、朱馨艺、邓　钢、王　龙）
Author(s):
CHEN Si-si;  ZHU Xin-yi;  DENG Gang;  WANG Long.
Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430060, China
关键词:
小胶质细胞; 长链非编码RNA; lncRNA LINC01089; miR-449c-5p; 小胶质细胞极化
Keywords:
Microglia;  Long non-coding RNA;  lnvRNA LINC01089;  miR-449c-5p;  Microglia polarization
分类号:
A
DOI:
10.13798/j.issn.1009-153X.2022.03.012
文献标志码:
R 743
摘要:
目的 探讨长链非编码RNA（lncRNA） LINC01089对小胶质细胞氧糖剥夺/复氧（OGD/R）损伤后表型和炎症反应的影响及其机制。方法 体外培养小鼠小胶质细胞B-V2细胞，OGD/R损伤后，转染lncRNA LINC01089过表达质粒及其空载质粒、沉默质粒siRNA及阴性对照，以及miR-449c-5p模拟物、抑制剂及其阴性对照寡核苷酸。采用qRT-PCR检测M1型小胶质细胞标记物iNOS mRNA和CD86 mRNA、M2型小胶质细胞标记物Arg1 mRNA和CD206 MRNA、lncRNA LINC01089、miR-449c-5p的表达水平；采用ELISA检测细胞上清液炎性因子的含量；采用免疫印迹法检测细胞STAT6蛋白表达水平。荧光素酶报告基因实验验证lncRNA LINC01089与miR-449c-5p以及miR-449c-5p与STAT6的靶向关系。结果 OGD/R损伤后，B-V2细胞lncRNA LINC01089表达水平显著下调，miR-449c-5p表达水平显著上调。荧光素酶报告基因实验结果显示，lncRNA LINC01089靶向负调控miR-449c-5p表达，miR-449c-5p靶向负调控STAT6表达。过表达lncRNA LINC01089，显著降低M1型标记物iNOS mRNA和CD86 mRNA表达水平及细胞上清液促炎因子IL-1β、IL-6和TNF-α的含量，明显增高M2型标记物Arg-1 mRNA和CD206 mRNA表达水平及细胞上清液抗炎因子IL-4、IL-10和TGF-β的含量。过表达lncRNA LINC01089可靶向负调控B-V2细胞miR-449c-5p表达，促进STAT6蛋白表达。结论 小胶质细胞OGD/R损伤后，lncRNA LINC01089表达下调，促使小胶质细胞向M1型转化，诱导炎症反应，其机制可能与靶向负调控miR-449c-5p/STAT6信号轴有关。
Abstract:
Objective To investigate the effect of lncRNA-LINC01089 on the phenotypic polarization of mouse microglia after oxygen-glucose deprivation/reoxygenation (OGD/R) injury. Methods The mouse microglia B-V2 cells were cultured in vitro. After OGD/R injury, lncRNA LINC01089 overexpression plasmid and its empty vector, silencing plasmid siRNA and negative control, as well as miR-449c-5p mimics, inhibitors and its negative control oligonucleotides were transfected into the cells. The expression levels of M1-type microglia markers iNOS and CD86 mRNAs, M2-type microglia markers Arg1 and CD206 mRNAs, lncRNA LINC01089, and miR-449c-5p were detected by qRT-PCR. The contents of inflammatory factors in cell supernatant were detected by ELISA. Western blotting was used to detect the expression level of STAT6 protein in B-V2 cells. The luciferase reporter gene experiment was used to verify the targeting relationship between lncRNA LINC01089 and miR-449c-5p, and miR-449c-5p and STAT6. Results After OGD/R injury, the expression level of lncRNA LINC01089 in B-V2 cells was significantly down-regulated, and the expression level of miR-449c-5p was significantly up-regulated. The results of the luciferase reporter gene assay showed that lncRNA LINC01089 targeted and negatively regulated the expression of miR-449c-5p, and miR-449c-5p targeted and negatively regulated the expression of STAT6. Overexpression of lncRNA LINC01089 significantly decreased the expression levels of M1-type markers iNOS and CD86 mRNAs, and significantly decreased the contents of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in the cell supernatant, and significantly increased the M2-type marker Arg-1 and CD206 mRNAs expression levels, and significantly increased the contents of anti-inflammatory factors IL-4, IL-10 and TGF-β in cell supernatants. Overexpression of lncRNA LINC01089 can target and negatively regulate the expression of miR-449c-5p in B-V2 cells and promote the expression of STAT6 protein. Conclusions After OGD/R injury, the expression of lncRNA LINC01089 is down-regulated in microglia, which promotes the transformation of microglia to M1 type and induces inflammatory response. The mechanism may be related to the negative regulation of miR-449c-5p/STAT6 signaling axis.

参考文献/References:

[1] Benjamin EJ, Blaha MJ, Chiuve SE, et al. Heart disease andstroke statistics-2017 update: a report from the american heart association [J]. Circulation, 2017, 135(10): e146-e603.
[2] Moskowitz MA, Lo EH, Iadecola C. The science of stroke: mechanisms in search of treatments [J]. Neuron, 2010, 67(2): 181-198.
[3] Fisher M, Saver JL. Future directions of acute ischaemic stroke therapy [J]. Lancet Neurol, 2015, 14(7): 758-767
[4] Wang Y, Pan WY, Ge JS, et al. A review of the relationship between long noncoding RNA and post-stroke injury repair [J]. J Int Med Res, 2019, 47(10): 4619-4624.
[5] Saugstad JA. Non-coding RNAs in stroke and neuroprotec-tion [J]. Front Neurol, 2015, 6: 50.
[6] Salter MW, Beggs S. Sublime microglia: expanding roles for the guardians of the CNS [J]. Cell, 2014, 158(1): 15-24.
[7] Patel AR, Ritzel R, McCullough LD, et al. Microglia and ischemic stroke: a double-edged sword [J]. Int J Physiol Pathophysiol Pharmacol, 2013, 5(2): 73-90.
[8] Ma Y, Wang J, Wang Y, et al. The biphasic function of mic-roglia in ischemic stroke [J]. Prog Neurobiol, 2017, 157: 247-272.
[9] Van Ginderachter JA, Movahedi K, Hassanzadeh GhassabehG, et al. Classical and alternative activation of mononuclear phagocytes: picking the best of both worlds for tumor pro-motion [J]. Immunobiology, 2006, 211(6-8): 487-501.
[10] Wang J, Zhao H, Fan Z, et al. Long noncoding RNA H19 promotes neuroinflammation in ischemic stroke by driving histone deacetylase 1-Dependent M1 microglial polarization[J]. Stroke, 2017, 48(8): 2211-2221.
[11] Ponomarev ED, Veremeyko T, Weiner HL. MicroRNAs are universal regulators of differentiation, activation, and pola-rization of microglia and macrophages in normal and disea-sed CNS [J]. Glia, 2013, 61(1): 91-103
[12] Ponomarev ED, Veremeyko T, Barteneva N, et al. MicroRNA-124 promotes microglia quiescence and suppresses EAE by deactivating macrophages via the C/EBP-α-PU.1 pathway [J]. Nat Med, 2011, 17(1): 64-70.
[13] Lawrence T, Natoli G. Transcriptional regulation of macro-phage polarization: enabling diversity with identity [J]. Nat Rev Immunol, 2011, 11(11): 750-761.
[14] Hou J, Schindler U, Henzel WJ, et al. An interleukin-4-induced transcription factor: IL-4 Stat [J]. Science, 1994, 265(5179): 1701-1706.
[15] Czimmerer Z, Daniel B, Horvath A, et al. The transcription factor STAT6 mediates direct repression of inflammatory enhancers and limits activation of alternatively polarized macrophages [J]. Immunity, 2018, 48(1): 75-90.
[16] Wang Q, Wei S, Zhou H, et al. Hyperglycemia exacerbates acetaminophen-induced acute liver injury by promoting liver-resident macrophage proinflammatory response via AMPK/PI3K/AKT-mediated oxidative stress [J]. Cell Death Discov, 2019, 5: 119.

相似文献/References:

[1]乔战东　刘雷震　贾利云　李经纶　刘　伟　曹付强　姚安会　王本瀚.人脑挫裂伤后早期小胶质细胞PD-L1的表达[J].中国临床神经外科杂志,2016,(04):216.[doi:10.13798/j.issn.1009-153X.2016.04.008]
　QIAO Zhan-dong,LIU Lei-zhen,et al.PD-L1 expression in microglial cells early after contusion and laceration of human brain[J].,2016,(03):216.[doi:10.13798/j.issn.1009-153X.2016.04.008]
[2]谈胤求　陈谦学　张红波　吴　鹏　李云涛　王栋良　袁凡恩.长链非编码RNA PVT1对人脑胶质瘤U251细胞增殖能力的影响[J].中国临床神经外科杂志,2016,(10):608.[doi:10.13798/j.issn.1009-153X.2016.10.013]
　TAN Yin-qiu,CHEN Qian-xue,ZHANG Hong-bo,et al.Effect of lncRNA-PVT1 on proliferation of glioma U251 cells[J].,2016,(03):608.[doi:10.13798/j.issn.1009-153X.2016.10.013]
[3]徐　阳　刘宝辉　袁凡恩　刘骏辉　许鹏飞　陈谦学.小胶质细胞在缺血性脑卒中中作用机制的研究进展[J].中国临床神经外科杂志,2018,(07):503.[doi:10.13798/j.issn.1009-153X.2018.07.020]
[4]宋维强 综述　林志国 审校.小胶质细胞与癫痫的关系研究进展[J].中国临床神经外科杂志,2019,(05):314.[doi:10.13798/j.issn.1009-153X.2019.05.021]
[5]陈　波　云亚滨　杜俊峰　王伟志.lncRNA SNHG16对胶质瘤U251细胞增殖、侵袭和迁移的影响[J].中国临床神经外科杂志,2020,(11):771.[doi:doi:10.13798/j.issn.1009-153X.2020.11.012]
　CHEN Bo,YUN Ya-bin,DU Jun-feng,et al.Effect of lncRNA SNHG16 on proliferation, invasion and migration of glioma U251 cells[J].,2020,(03):771.[doi:doi:10.13798/j.issn.1009-153X.2020.11.012]
[6]孙青青　任洪波　牛国栋.lncRNA SNHG16在人脑胶质瘤组织中的表达及其与病人预后的关系[J].中国临床神经外科杂志,2020,(12):831.[doi:10.13798/j.issn.1009-153X.2020.12.005]
　SUN Qing-qing,REN Hong-bo,NIU Guo-dong..Expression of lncRNA SNHG16 in human glioma tissues and its relationship with prognosis of glioma patients[J].,2020,(03):831.[doi:10.13798/j.issn.1009-153X.2020.12.005]
[7]罗孝全　唐　辉　冯　浩　陈　兵　孙　谋.lncRNA MIR4435-2HG的表达水平及甲基化状态与脑胶质瘤病理分级的关系[J].中国临床神经外科杂志,2021,26(01):26.[doi:10.13798/j.issn.1009-153X.2021.01.009]
　LUO Xiao-quan,TANG Hui,FENG Hao,et al.Relationship between expression level and methylation status of lncRNA MIR4435-2HG and pathological grade of gliomas[J].,2021,26(03):26.[doi:10.13798/j.issn.1009-153X.2021.01.009]
[8]郑江林　王　旋　汪珉杰　姜晓兵.人脑胶质瘤lncRNA PIK3CD-AS2表达及临床意义[J].中国临床神经外科杂志,2021,26(09):687.[doi:10.13798/j.issn.1009-153X.2021.09.011]
　ZHENG Jiang-lin,WANG Xuan,WANG Min-jie,et al.Expression of lncRNA PIK3CD-AS2 in human brain gliomas and its clinical significance[J].,2021,26(03):687.[doi:10.13798/j.issn.1009-153X.2021.09.011]
[9]杨峰彪　张祎年.小胶质细胞在癫痫中作用的研究进展[J].中国临床神经外科杂志,2021,26(09):729.[doi:10.13798/j.issn.1009-153X.2021.09.027]
[10]李占成　石　铁　周卫忠.胶质瘤自噬相关长链非编码RNA的生信分析[J].中国临床神经外科杂志,2021,26(11):839.[doi:10.13798/j.issn.1009-153X.2021.11.007]
　LI Zhan-cheng,SHI Tie,ZHOU Wei-zhong.Autophagy-related long non-coding RNA signature in glioma patients based on bioinformatics analysis[J].,2021,26(03):839.[doi:10.13798/j.issn.1009-153X.2021.11.007]

备注/Memo

备注/Memo:
通讯作者：王　龙，E-mail：361140072@qq.com
更新日期/Last Update: 1900-01-01