Effects of Siglec-E on parthenolide inhibiting microglia M1 polarization and targeting MAPK/NF-κB pathway

Wenying FU; Pengbo WANG; Xiangyu TANG; Liyuan FENG; Yuting HUANG; Peng LI

doi:10.16016/j.2097-0927.202312110

您当前的位置：

首页 >

文章列表页 >

Effects of Siglec-E on parthenolide inhibiting microglia M1 polarization and targeting MAPK/NF-κB pathway

Neuroscience | 更新时间：2026-01-09

- Effects of Siglec-E on parthenolide inhibiting microglia M1 polarization and targeting MAPK/NF-κB pathway
- Journal of Army Medical University Vol. 46, Issue 7, Pages: 651-660(2024)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系生药学与中药学教研室
  2.400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系药剂学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81673098)
- DOI：10.16016/j.2097-0927.202312110
  CLC： R285.5;R322.8;R394.3
- Received：21 December 2023，
  
  Revised：2024-01-05，
  
  Published：15 April 2024
- 稿件说明：
移动端阅览
Wenying FU, Pengbo WANG, Xiangyu TANG, et al. Effects of Siglec-E on parthenolide inhibiting microglia M1 polarization and targeting MAPK/NF-κB pathway[J]. Journal of Army Medical University, 2024, 46(7): 651-660.
DOI：

Wenying FU, Pengbo WANG, Xiangyu TANG, et al. Effects of Siglec-E on parthenolide inhibiting microglia M1 polarization and targeting MAPK/NF-κB pathway[J]. Journal of Army Medical University, 2024, 46(7): 651-660. DOI： 10.16016/j.2097-0927.202312110.

摘要

目的

探讨唾液酸结合样凝集素E(sialic acid binding lectin E，Siglec-E)对小白菊内酯(parthenolide，PTL)抑制脂多糖(lipopolysaccharide，LPS)诱导的小鼠小胶质细胞(BV2)M1极化作用的影响。

方法

① 从基因表达综合数据库(Gene Expression Omnibus，GEO)获取

Siglece

相关小鼠脑组织单细胞测序数据，分为野

生型组(

，

=3)和敲除型组(

Siglece

-/-

，

=4)。从中筛选出小胶质细胞并分析相关差异基因及通路富集。通过shRNA干扰技术构建BV2细胞株模型，得到

-shRNA BV2和

Siglece

-shRNA BV2细胞并检测Siglec-E(

Siglece

)表达水平。②将

-shRNA和

Siglece

-shRNA细胞分别分为Control组、LPS组、PTL组和PTL+LPS组(

=3)，采用RT-qPCR检测小胶质细胞M1极化标志物

iNOS

、

-1

和

-6的mRNA水平。将

Siglece

和

cre

小鼠交配，获得小胶质细胞特异性Siglec-E缺失(

Siglece

cre

)小鼠，并建立LPS诱导的神经炎症模型。③将出生4 d的各9只

和

Siglece

cre

雄鼠分成Control组、LPS组和PTL+LPS组(

=3)。采用RT-qPCR、免疫荧光技术和Western blot验证敲除效果以及极化相关通路，研究Siglec-E影响PTL抑制小胶质细胞M1极化的机制。

结果

与

-shRNA组比较，

Siglece

-shRNA组中Siglec-E表达显著降低(

0.01)，说明SiglecE敲除细胞模型建立成功。在LPS刺激的条件下，与Control组比较，

-shRNA细胞和

Siglece

-shRNA细胞的

iNOS

、

-1

和

-6的mRNA水平均显著上调(

0.01)。PTL与LPS共同作用下，

-shRNA细胞中上述M1极化的标志物均明显下降(

0.05)。而对于

Siglece

-shRNA细胞，上述M1极化的标志物无显著下调。PTL对BV2细胞JNK、IκB蛋白的磷酸化具有抑制作用(

0.01)，也可抑制NF-κB核转位，而下调Siglec-E则会减弱该作用。与

组小鼠比较，

Siglece

cre

小鼠小胶质细胞Siglec-E表达显著降低(

0.01)，PTL对

Siglece

cre

小鼠小胶质细胞NF-κB磷酸化水平的抑制作用也降低。

结论

小胶质细胞Siglec-E缺失会减弱PTL靶向MAPK/NF-κB通路对其M1极化的抑制作用。

Abstract

Objective

To explore the impact of the sialic acid binding lectin-E (Siglec-E) on the inhibitory properties of parthenolide(PTL) against lipopolysaccharide (LPS)-induced M1 polarization of microglia(BV2).

Methods

① Single cell sequencing data of

Siglece

related mouse brain tissue was obtained from Gene Expression Omnibus (GEO) database and divided into the

group (

=3) and the

Siglece

-/-

group (

=4). The microglia cells were screened

and the enrichment analysis was performed to analyze related differential genes and pathways. BV2 cells were constructed by the shRNA interference technique and were divided into

-shRNA and

Siglece

-shRNA to detect the expression level of Siglec-E (

Siglece

). ②

-shRNA and

Siglece

-shRNA cells were respectively divided into the Control group

LPS group

PTL group and PTL+LPS group (

=3). The mRNA levels of markers of M1 polarization in microglia

iNOS

-1

and

-6

were detected by RT-qPCR.

Siglece

and

cre

mice were mated to obtain microglia-specific

Siglece

deletion (

Siglece

cre

) mice

and LPS-induced neuroinflammation model was established. ③ Nine

and

Siglece

cre

male mice were assigned to the Control group

LPS group and PTL+LPS group (

=3). RT-qPCR

immunofluorescence assay and Western blotting were used to verify the knock-out effect and polarization-related pathways

and to investigate the mechanism of Siglec-E affecting PTL inhibition of M1 polarization of microglia.

Results

Compared with the

-shRNA group

the expression of Siglec-E in the

Siglece

-shRNA group was significantly decreased (

0.01)

indicating that the Siglec-E knock-down cell model was successfully established. With the stimulation of LPS

mRNA levels of

iNOS

-1

and

-6 were significantly up-regulated compared with the Control group both in shRNA cells and

Siglece

-shRNA cells (

0.01). With the influence of PTL and LPS

the markers of M1 polarization in

-shRNA cells mentioned before were significantly decreased (

0.05)

while for

Siglice

-shRNA cells

there were no significant changes in the markers of M1 polarization. PTL inhibited the phosphorylation of JNK and IκB protein (

0.01) and the nuclear translocation of NF-κB in BV2 cells

down-regulated Siglec-E

and weakened the inhibitory effect. Compared with mice in the

group

the expression of Siglec-E in microglia of

Siglece

cre

mice was decreased significantly (

0.01)

and the inhibitory effect of PTL on the phosphorylation of NF-κB in microglia of

Siglece

cre

mice was also decreased.

Conclusion

The absence of Siglec-E in microglia attenuates the inhibition of M1 polarization by the MAPK/NF-κB pathway targeted by PTL.

关键词

Keywords

references

STOESSEL M B, MAJEWSKA A K. Little cells of the little brain: microglia in cerebellar development and function[J]. Trends Neurosci, 2021, 44(7): 564-578. DOI: 10.1016/j.tins.2021.04.001.

WANG L T, LIU Q, ZHANG Y L, et al . Establishment of BV2 microglia polarization model and its effect on Toxoplasma gondii proliferation[J ] . Res Vet Sci, 2019, 125: 382-389. DOI: 10.1016/j.rvsc.2019.08.002.

RANSOHOFF R M. A polarizing question: do M1 and M2 microglia exist?[J]. Nat Neurosci, 2016, 19: 987-991. DOI: 10.1038/nn.4338.

ZHANG B, WEI Y Z, WANG G Q, et al . Targeting MAPK pathways by naringenin modulates microglia M1/M2 polarization in lipopolysaccharide-stimulated cultures[J ] . Front Cell Neurosci, 2019, 12: 531. DOI: 10.3389/fncel.2018.00531.

GUO S R, WANG H, YIN Y F. Microglia polarization from M1 to M2 in neurodegenerative diseases[J]. Front Aging Neurosci, 2022, 14: 815347. DOI: 10.3389/fnagi.2022.815347.

ZHOU X Q, ZHAO S Y, LIU T T, et al . Schisandrol A protects AGEs-induced neuronal cells death by allosterically targeting ATP6V0d1 subunit of V-ATPase[J ] . Acta Pharm Sin B, 2022, 12(10): 3843-3860. DOI: 10.1016/j.apsb.2022.06.013.

WANG Q Q, HAN B, MAN X, et al . Chuanzhitongluo regulates microglia polarization and inflammatory response in acute ischemic stroke[J ] . Brain Res Bull, 2022, 190: 97-104. DOI: 10.1016/j.brainresbull.2022.09.015.

GUO Y T, DAI W B, ZHENG Y, et al . Mechanism and regulation of microglia polarization in intracerebral hemorrhage [J ] . Molecules, 2022, 27(20): 7080. DOI: 10.3390/molecules27207080.

TAO G J, QIAN D F, LI L W, et al . Parthenolide promotes the repair of spinal cord injury by modulating M1/M2 polarization via the NF-κB and STAT 1/3 signaling pathway[J ] . Cell Death Discov, 2020, 6(1): 97. DOI: 10.1038/s41420-020-00333-8..

MAGNI P, RUSCICA M, DOZIO E, et al . Parthenolide inhibits the LPS-induced secretion of IL-6 and TNF-α and NF-κB nuclear translocation in BV-2 microglia[J ] . Phytother Res, 2012, 26(9): 1405-1409. DOI: 10.1002/ptr.3732.

ZHAO Z J, XIANG J Y, LIU L, et al . Parthenolide, an inhibitor of the nuclear factor-κB pathway, ameliorates dextran sulfate sodium-induced colitis in mice[J ] . Int Immunopharmacol, 2012, 12(1): 169-174. DOI: 10.1016/j.intimp.2011.11.007.

CLAUDE J, LINNARTZ-GERLACH B, KUDIN A P, et al . Microglial CD33-related Siglec-E inhibits neurotoxicity by preventing the phagocytosis-associated oxidative burst[J ] . J Neurosci, 2013, 33(46): 18270-18276. DOI: 10.1523/JNEUROSCI.2211-13.2013.

SIDDIQUI S S, MATAR R, MERHEB M, et al . Siglecs in brain function and neurological disorders[J ] . Cells, 2019, 8(10): 1125. DOI: 10.3390/cells8101125.

BLOCK M L, ZECCA L, HONG J S. Microglia-mediated neurotoxicity: uncovering the molecular mechanisms[J]. Nat Rev Neurosci, 2007, 8(1): 57-69. DOI: 10.1038/nrn2038.

LI L X, CHEN Y, SLUTER M N, et al . Ablation of Siglec-E augments brain inflammation and ischemic injury[J ] . J Neuroinflammation, 2022, 19(1): 191. DOI: 10.1186/s12974-022-02556-1.

TANG J R, MICHAELIS K A, NOZIK-GRAYCK E, et al . The NF-κB inhibitory proteins IκBα and IκBβ mediate disparate responses to inflammation in fetal pulmonary endothelial cells[J ] . J Immunol, 2013, 190(6): 2913-2923. DOI: 10.4049/jimmunol.1202670.

TENTILLIER N, ETZERODT A, OLESEN M N, et al . Anti-inflammatory modulation of microglia via CD163-targeted glucocorticoids protects dopaminergic neurons in the 6-OHDA Parkinson's disease model[J ] . J Neurosci, 2016, 36(36): 9375-9390. DOI: 10.1523/JNEUROSCI.1636-16.2016..

DAI Y J, WEI T F, SHEN Z X, et al . Classical HDACs in the regulation of neuroinflammation[J ] . Neurochem Int, 2021, 150: 105182. DOI: 10.1016/j.neuint.2021.105182.

XIE J H, TUO P, ZHANG W, et al . Inhibition of the TLR4/NF-κB pathway promotes the polarization of LPS-induced BV2 microglia toward the M2 phenotype[J ] . Neuroreport, 2023, 34(17): 834-844. DOI: 10.1097/WNR.0000000000001961.

FAN M D, WANG C, ZHAO X Y, et al . Parthenolide alleviates microglia-mediated neuroinflammation via MAPK/TRIM31/NLRP3 signaling to ameliorate cognitive disorder[J ] . Int Immunopharmacol, 2023, 120: 110287. DOI: 10.1016/j.intimp.2023.110287..

LINNARTZ-GERLACH B, KOPATZ J, NEUMANN H. Siglec functions of microglia[J]. Glycobiology, 2014, 24(9): 794-799. DOI: 10.1093/glycob/cwu044.

LIU J Y, GENG X F, HOU J X, et al . New insights into M1/M2 macrophages: key modulators in cancer progression[J ] . Cancer Cell Int, 2021, 21(1): 389. DOI: 10.1186/s12935-021-02089-2.

MA X R, LIU H F, ZHU J, et al . MiR-185-5p regulates inflammation and phagocytosis through CDC42/JNK pathway in macrophages[J ] . Genes, 2022, 13(3): 468. DOI: 10.3390/genes1303 0468.

THIESLER H, BEIMDIEK J, HILDEBRANDT H. Polysialic acid and Siglec-E orchestrate negative feedback regulation of microglia activation[J]. Cell Mol Life Sci, 2021, 78(4): 1637-1653. DOI: 10.1007/s00018-020-03601-z.

Views

479

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Neferine attenuates parkinson’s disease via modulating microglial pyroptosis mediated by ROS/NLRP3/Caspase-1 signaling pathway

SOCS3 ameliorates Parkinson’s disease neuropathology by suppressing NF-κB signaling-mediated microglial inflammation

FDFT1 inhibits macrophage M1 polarization and promotes colorectal cancer progression

Protective effects of parthenolide on intestinal barrier function in septic rats

Effects of running on medial prefrontal cortex and microglia in APP/PS1 mice

Related Author

LIU Fenfen

FENG Dan

LIU Yun

ZUO Fangya

GUO Xiuhong

LIU Yuqi

CHEN Lanlan

WANG Yujie

Related Institution

Department of General Practice， Guizhou Provincial People’s Hospital

Graduate School， Zunyi Medical University

Department of Critical Care Medicine， the First People’s Hospital of Qingzhen City

Graduate School， Guizhou University of Traditional Chinese Medicine

Department of Critical Care Medicine， Qingzhen First People’s Hospital

⁰