Hypoxia-downregulated Siglec-9 glycan ligands induce abnormal activation of platelet

Yuanting SHE; Dongfeng ZENG; Hongmei LIU; Niting WU; Yi JIA

doi:10.16016/j.2097-0927.202111025

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Hypoxia-downregulated Siglec-9 glycan ligands induce abnormal activation of platelet

Basic Medicine | 更新时间：2026-01-09

- Hypoxia-downregulated Siglec-9 glycan ligands induce abnormal activation of platelet
- Journal of Army Medical University Vol. 44, Issue 6, Pages: 541-548(2022)
- 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：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81673427);the Innovation Project of Army Medical Center of PLA(2019CXJSB020)
- DOI：10.16016/j.2097-0927.202111025
  CLC： R331.143;R363.21;R364.4
- Received：03 November 2021，
  
  Revised：2021-12-03，
  
  Published：30 March 2022
- 稿件说明：
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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：

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

references

邱义稳, 宋航, 赵婕, 等. 高原环境血小板的变化及其作用机制探讨[J]. 西南国防医药, 2015, 25(10): 1146-1148. DOI: 10.3969/j.issn.1004-0188.2015.10.041.

QIU Y W, SONG H, ZHAO J, et al . The changes of platelets in high altitude environment and the mechanism of action[J ] . Med J Natl Defend Forces Southwest China, 2015, 25(10): 1146-1148. DOI: 10.3969/j.issn.1004-0188.2015.10.041.

周建丽, 王斌, 谢祥红, 等. 不同海拔高原驻训人员外周血小板参数变化及意义[J]. 武警医学院学报, 2010, 19(4): 304-305.

ZHOU J L, WANG B, XIE X H, et al . Changes of peripheral platelet count in training personnel at different altitude[J ] . Acta Acad Med CPAF, 2010, 19(4): 304-305.

柯景彬, 晋军, 黄岚, 等. 急性高原暴露及短期习服对青年男性血小板相关参数的影响[J]. 解放军医学杂志, 2018, 43(3): 251-256. DOI: 10.11855/j.issn.0577-7402.2018.03.13.

KE J B, JIN J, HUANG L, et al . Influence of acute high altitude exposure and short-term acclimation on platelet-associated parameters in healthy young man[J ] . Med J Chin PLA, 2018, 43(3): 251-256. DOI: 10.11855/j.issn.0577-7402.2018.03.13.

SHANG C X, WUREN T N, GA Q, et al . The human platelet transcriptome and proteome is altered and pro-thrombotic functional responses are increased during prolonged hypoxia exposure at high altitude[J ] . Platelets, 2020, 31(1): 33-42. DOI: 10.1080/09537104.2019.1572876.

秦芳梅, 叶洁瑜, 黄顺桦, 等. 血小板活化功能及血小板微粒对重型血友病A临床异质性的意义[J]. 第三军医大学学报, 2015, 37(13): 1362-1366. DOI: 10.16016/j.1000-5404.201504095.

QIN F M, YE J Y, HUANG S H, et al . Significance of platelet activating capacity and platelet-derived microparticles in clinical phenotype heterogeneity of hemophlia A: report of 31 cases[J ] . J Third Mil Med Univ, 2015, 37(13): 1362-1366. DOI: 10.16016/j.1000-5404.201504095.

MACAULEY M S, CROCKER P R, PAULSON J C. Siglec-mediated regulation of immune cell function in disease[J]. Nat Rev Immunol, 2014, 14(10): 653-666. DOI: 10.1038/nri3737.

UCHIYAMA S, SUN J, FUKAHORI K, et al . Dual actions of group B Streptococcus capsular sialic acid provide resistance to platelet-mediated antimicrobial killing[J ] . Proc Natl Acad Sci USA, 2019, 116(15): 7465-7470. DOI: 10.1073/pnas.1815572116.

KORPORAAL S J, KOEKMAN C A, VERHOEF S, et al . Downregulation of platelet responsiveness upon contact with LDL by the protein-tyrosine phosphatases SHP-1 and SHP-2[J ] . Arterioscler Thromb Vasc Biol, 2009, 29(3): 372-379. DOI: 10.1161/ATVBAHA.108.173278.

LIZCANO A, SECUNDINO I, DÖHRMANN S, et al . Erythrocyte sialoglycoproteins engage Siglec-9 on neutrophils to suppress activation[J ] . Blood, 2017, 129(23): 3100-3110. DOI: 10.1182/blood-2016-11-751636.

DELAVERIS C S, CHIU S H, RILEY N M, et al . Modulation of immune cell reactivity with cis-binding Siglec agonists[J ] . Proc Natl Acad Sci USA, 2021, 19, 118(3): e2012408118. DOI: 10.1073/pnas.2012408118.

JIA Y, YU H F, FERNANDES S M, et al . Expression of ligands for Siglec-8 and Siglec-9 in human airways and airway cells[J ] . J Allergy Clin Immunol, 2015, 135(3): 799-810.e7. DOI: 10.1016/j.jaci.2015.01.004.

ZHANG Y X, ZHENG Y, LI J, et al . Immunoregulatory Siglec ligands are abundant in human and mouse aorta and are up-regulated by high glucose[J ] . Life Sci, 2019, 216: 189-199. DOI: 10.1016/j.lfs.2018.11.049.

HEBER S, VOLF I. Effects of physical (in)activity on platelet function[J]. Biomed Res Int, 2015, 2015: 165078. DOI: 10.1155/2015/165078.

TOMAIUOLO M, BRASS L F, STALKER T J. Regulation of platelet activation and coagulation and its role in vascular injury and arterial thrombosis[J]. Interv Cardiol Clin, 2017, 6(1): 1-12. DOI: 10.1016/j.iccl.2016.08.001.

STANCZAK M A, SIDDIQUI S S, TREFNY M P, et al . Self-associated molecular patterns mediate cancer immune evasion by engaging Siglecs on T cells[J ] . J Clin Invest, 2018, 128(11): 4912-4923. DOI: 10.1172/jci120612.

MA P S, CIERNIEWSKA A, SIGNARVIC R, et al . A newly identified complex of spinophilin and the tyrosine phosphatase, SHP-1, modulates platelet activation by regulating G protein-dependent signaling[J ] . Blood, 2012, 119(8): 1935-1945. DOI: 10.1182/blood-2011-10-387910.

COXON C H, SADLER A J, HUO J, et al . An investigation of hierachical protein recruitment to the inhibitory platelet receptor, G6B-B[J ] . PLoS One, 2012, 7(11): e49543. DOI: 10.1371/journal.pone.0049543.

HSU Y W, HSU F F, CHIANG M T, et al . Siglec-E retards atherosclerosis by inhibiting CD36-mediated foam cell formation[J ] . J Biomed Sci, 2021, 28(1): 5. DOI: 10.1186/s12929-020-00698-z.

谢玉海, 王学军. 高原地区患者术前凝血功能特征及血栓弹力图评估凝血功能的价值研究[J]. 中国现代医学杂志, 2017, 27(17): 80-83. DOI: 10.3969/j.issn.1005-8982.2017.14.017.

XIE Y H, WANG X J. Preoperative coagulation function of patients in plateau area and value of thrombelastography in assessment of coagulative function[J]. China J Mod Med, 2017, 27(17): 80-83. DOI: 10.3969/j.issn.1005-8982.2017.14.017.

LIU H, ZHENG Y, ZHANG Y, et al . Immunosuppressive Siglec-E ligands on mouse aorta are up-regulated by LPS via NF-κB pathway[J ] . Biomed Pharmacother, 2020, 122: 109760. DOI: 10.1 016/j.biopha.2019.109760.

KISER Z M, LIZCANO A, NGUYEN J, et al . Decreased erythrocyte binding of Siglec-9 increases neutrophil activation in sickle cell disease[J ] . Blood Cells Mol Dis, 2020, 81: 102399. DOI: 10.1016/j.bcmd.2019.102399.

WANDALL H H, RUMJANTSEVA V, SØRENSEN A L, et al . The origin and function of platelet glycosyltransferases[J ] . Blood, 2012, 120(3): 626-635. DOI: 10.1182/blood-2012-02-409235.

CHEN G Y, BROWN N K, WU W, et al . Broad and direct interaction between TLR and Siglec families of pattern recognition receptors and its regulation by Neu1[J ] . eLife, 2014, 3: e04066. DOI: 10.7554/eLife.04066.

ZHANG L, WEI T T, LI Y, et al . Functional metabolomics characterizes a key role for N-acetylneuraminic acid in coronary artery diseases[J ] . Circulation, 2018, 137(13): 1374-1390. DOI: 10.1161/circulationaha.117.031139.

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