缺氧下调血小板表面Siglec-9聚糖配体导致血小板功能异常活化

摄渊婷; 曾东风; 刘红梅; 吴妮婷; 贾乙

doi:10.16016/j.2097-0927.202111025

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缺氧下调血小板表面Siglec-9聚糖配体导致血小板功能异常活化

基础医学 | 更新时间：2026-01-09

- 缺氧下调血小板表面Siglec-9聚糖配体导致血小板功能异常活化
- Hypoxia-downregulated Siglec-9 glycan ligands induce abnormal activation of platelet
- 陆军军医大学学报 2022年44卷第6期页码：541-548
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系药剂学教研室
  2.400042 重庆，陆军特色医学中心血液科
- 作者简介：
  
  [贾乙，E-mail：jy@tmmu.edu.cn JIA Yi, E-mail: jy@tmmu.edu.cn
- 基金信息：
  
  国家自然科学基金面上项目(81673427);陆军特色医学中心创新项目(2019CXJSB020)
- DOI：10.16016/j.2097-0927.202111025
  中图分类号： R331.143;R363.21;R364.4
- 收稿：2021-11-03，
  
  修回：2021-12-03，
  
  纸质出版：2022-03-30
- 稿件说明：
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摄渊婷, 曾东风, 刘红梅, 等. 缺氧下调血小板表面Siglec-9聚糖配体导致血小板功能异常活化[J]. 陆军军医大学学报, 2022,44(6):541-548.

Yuanting SHE, Dongfeng ZENG, Hongmei LIU, et al. Hypoxia-downregulated Siglec-9 glycan ligands induce abnormal activation of platelet[J]. Journal of Army Medical University, 2022, 44(6): 541-548.
摄渊婷, 曾东风, 刘红梅, 等. 缺氧下调血小板表面Siglec-9聚糖配体导致血小板功能异常活化[J]. 陆军军医大学学报, 2022,44(6):541-548. DOI： 10.16016/j.2097-0927.202111025.

Yuanting SHE, Dongfeng ZENG, Hongmei LIU, et al. Hypoxia-downregulated Siglec-9 glycan ligands induce abnormal activation of platelet[J]. Journal of Army Medical University, 2022, 44(6): 541-548. DOI： 10.16016/j.2097-0927.202111025.

摘要

目的

探讨血小板表面Siglec-9聚糖配体表达水平在缺氧条件下的变化及其对血小板活化功能的影响。

方法

鉴定血小板表面表达的Siglec-9/E聚糖配体，并加入唾液酸酶处理血小板，分为对照组和唾液酸酶组；对24只6~8周龄雄性C57BL/6小鼠和血小板进行缺氧处理，分为对照3 d组、缺氧3 d组、对照7 d组和缺氧7 d组(

=6)；将健康志愿者全血分离提取血小板放置于缺氧箱，分为对照组、缺氧48 h组、唾液酸神经节甘脂(Ganglioside GT1b

GT1b)组和奥司他韦组(

=3)。采用流式细胞术检测血小板上Siglec-9/E配体、神经氨酸酶1(Neuraminidase-1)等表达，以及二磷酸腺苷(ADP)、胶原和凝血酶刺激后血小板活化标记物CD62p的表达水平；利用髓系细胞条件性6只6~8周龄雄性Siglec-E基因敲除小鼠(Lyz2-cre：Siglec-E

flox/flox

)血小板验证Siglec-E对血小板活化的抑制作用。

结果

相对分子质量约为130×10

的Siglec-9/E配体在血小板表面广泛表达且具有唾液酸末端的聚糖结构；与对照7 d组相比，缺氧7 d组小鼠外周血血小板上该配体表达显著降低18.03%(

0.05)，CD62p表达在ADP、胶原和凝血酶刺激下显著增高(

0.01)，缺氧48 h组培养的血小板上该聚糖配体出现了相同改变(

0.01)；与野生型比较，敲基因小鼠血小板在ADP、胶原和凝血酶刺激下CD62p水平显著增高(

0.01)；Siglec-9/E特异性配体GT1b处理能够逆转缺氧导致的血小板异常活化(

0.05)；神经氨酸酶抑制剂能够阻断缺氧导致的Siglec-9配体表达降低(

0.05)。

结论

缺氧激活NEU1降低Siglec-9/E聚糖配体表达，减弱了Siglec-9/E途径抑制血小板活化的能力，导致血小板功能异常活化。

Abstract

Objective

To investigate the expression of sialic acid-binding immunoglobulin-like lectin-9 (Siglec-9)/E ligands in hypoxic condition and its effect on platelet activation.

Methods

The distribution

molecular size and glycoprotein class of Siglec-9 ligands on platelet were determined. The platelets were divided into control group and neuraminidase group [treated with α2-3

8 neuraminidase (NEU1)

]

. Twenty-four C57BL/6 male mice (6~8 weeks old) or cultured platelets were treated with or without hypoxia

and then the mice were divided into control 3-day group

hypoxia 3-day group

control 7-day group and hypoxia 7-day group

with 6 mice in each group. The platelets were separated and extracted from the whole blood of healthy volunteers and were placed in a hypoxia chamber

and then divided into control group

hypoxia 48-hour group

GT1b group and oseltamivir group (

=3). The expression of Siglec-E/9 ligands

NEU1

and CD62p which was activated by adenosine diphosphate (ADP)

collagen and thrombin on the platelet were detected by flow cytometry. The inhibitory effect of Siglec-E/9 ligands was verified using conditional knockout mice Lyz2-cre: Siglec-E

flox/flox

(male

6~8 weeks old

=6).

Results

Siglec-9 ligands are sialoglycoproteins with molecular weights of about 130×10

and are distributed broadly on the membrane of platelet.

In vivo

the expression of Siglec-E ligands was decreased by 18.03% in response to hypoxia treatment (

0.05)

and the stimulation of ADP

collagen and thrombin resul

ted in obviously elevated CD62p level in the platelets from the hypoxia 7-day group than those from the control 7-day group (

0.01). Similar Results were observed in the platelets of the hypoxia 48-hour group (

0.01). Compared with the platelets from wild-type mice

the platelets from the knockdown mice had significantly higher expression of CD62p after the stimulation of ADP

collagen and thrombin (

0.01). GT1b treatment could reverse the abnormal platelet activation induced by hypoxia (

0.05)

and treatment of oseltamivir

NEU1 inhibitor could block the decreased expression of Siglec-9 ligand caused by hypoxia (

0.05).

Conclusion

Hypoxia can decrease the expression of Siglec-9/E ligands

reduce Siglec-9/E-mediated platelet activation

and thus lead to abnormal activation of platelets

which is associated with the increased expression of NEU1.

关键词

Keywords

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