CFTR Regulates Platelet Activation via the P2Y<sub>12</sub>-Mediated Signaling Pathway

LI Jun; YANG Han-yan; HAN Hui; LI Mei; WANG Guan-lei

doi:10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0406

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CFTR Regulates Platelet Activation via the P2Y₁₂-Mediated Signaling Pathway

Basic Research | 更新时间：2024-02-19

- CFTR Regulates Platelet Activation via the P2Y₁₂-Mediated Signaling Pathway
- Journal of Sun Yat-sen University(Medical Sciences) Vol. 42, Issue 4, Pages: 521-527(2021)
- 作者机构：
  
  1.中山大学中山医学院药理学教研室和心脑血管研究中心，广东广州 510080
  2.中山大学附属第三医院特诊医疗病区，广东广州 510630
- 作者简介：
  
  LI Mei， E-mail： limei36@mail.sys.edu.cn
  WANG Guan-lei， E-mail： wangglei@mail.sysu.edu.cn
- 基金信息：
- DOI：10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0406
  CLC： R96
- Received：08 February 2021，
  
  Published：20 July 2021
- 稿件说明：
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李俊,杨涵滟,韩慧等.CFTR通过调控二磷酸腺苷受体P2Y₁₂影响血小板活化[J].中山大学学报（医学科学版）,2021,42(04):521-527.

LI Jun,YANG Han-yan,HAN Hui,et al.CFTR Regulates Platelet Activation via the P2Y₁₂-Mediated Signaling Pathway[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(04):521-527.
李俊,杨涵滟,韩慧等.CFTR通过调控二磷酸腺苷受体P2Y₁₂影响血小板活化[J].中山大学学报（医学科学版）,2021,42(04):521-527. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0406.

LI Jun,YANG Han-yan,HAN Hui,et al.CFTR Regulates Platelet Activation via the P2Y₁₂-Mediated Signaling Pathway[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(04):521-527. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0406.

摘要

目的

二磷酸腺苷受体P2Y

介导的血小板活化是血小板活化及血栓形成的核心机制，本研究探讨囊性纤维化跨膜转导调节子（CFTR）对P2Y

介导的血小板活化的影响。

方法

应用8~16周龄

CFTR

全基因敲除（

Cftr

-/-

）小鼠及野生型（

Cftr

+/+

）小鼠，分别制备胶原蛋白和肾上腺素诱导的肺栓塞小鼠模型。分离

Cftr

-/-

和

Cftr

+/+

小鼠外周血的血小板，采用免疫印迹技术检测P2Y

、PI3K以及AKT 总蛋白及磷酸化蛋白的表达。采用二磷酸腺苷（ADP）孵育人巨核细胞株Meg-01，检测ADP对CFTR和上述蛋白表达的影响。进一步应用特异性CFTR氯通道抑制剂CFTRinh-172与Meg-01细胞共孵育，检测其对CFTR和上述蛋白表达的影响。

结果

与

Cftr

+/+

小鼠肺栓塞模型组相比，

Cftr

-/-

鼠肺栓塞组织切片栓塞显著增加。

Cftr

-/-

显著增加小鼠外周血血小板P2Y

、磷酸化PI3K和AKT 蛋白表达。

P2Y

全基因敲除小鼠血小板CFTR蛋白表达无明显改变，上述结果提示CFTR负性调控P2Y

维持正常血小板活化功能。Meg-01细胞实验结果表明 ADP浓度依赖性地诱导Meg-01细胞P2Y

蛋白和PI3K以及AKT蛋白磷酸化增高，而CFTR蛋白表达水平逐渐降低。CFTR 特异性阻断剂CFTRinh-172显著诱导Meg-01细胞P2Y

蛋白表达增高。

结论

本研究揭示了血小板活化的一个新机制，即CFTR通过调控血小板P2Y

蛋白表达影响血小板活化。

Abstract

Objective

Platelet activation mediated by adenosine diphosphate receptor P2Y

is critical for platelet activation and thrombosis formation. The goal of the presented study was to investigate the role of CFTR in platelet activation mediated by P2Y

Methods

8-week-old

CFTR

knockout （

Cftr

-/-

） mice and age-matched wild-type （

Cftr

+/+

） mice were used to set up collagen and epinephrine-induced pulmonary embolism mouse models. The lung sections were prepared to observe the pulmonary embolism. The circulating platelets were isolated from peripheral blood of

Cftr

-/-

and

Cftr

+/+

mice， and were used to detect protein expressions of P2Y

， p-PI3K and p-AKT by western blot analysis. The above signaling proteins were also detected in human-derived megakaryocyte cell strains （Meg-01） treated with different concentration of adenosine diphosphate （ADP）. CFTRinh-172， a selective CFTR chloride channel blocker was also applied to observe how CFTR affected the protein level of P2Y

in Meg-01 cells.

Results

In the pulmonary embolism model， the embolization area in lung tissue sections from

Cftr

-/-

mice were significantly larger than that of

Cftr

+/+

mice. Compared with

Cftr

+/+

mice， the protein expression of P2Y

， p-PI3K and p-AKT in circulating platelets of

Cftr

-/-

mice was significantly elevated. However， no change was found in the protein expression of platelet CFTR in

P2Y

knockout mice， which suggested that CFTR might be in the upstream of P2Y

in the regulating mechanism of platelet activation. In Meg-01 cells， ADP concentration-dependently induced P2Y

protein expression but decreased CFTR expression. The incubation of Meg-01 cells with CFTR

Inh

-172 could significantly increase P2Y

expression.

Conclusions

This study revealed a new mechanism of platelet activation， that is， CFTR may regulate platelet activation by regulating P2Y

-mediated signaling pathway.

关键词

Keywords

references

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CFTR Regulates Platelet Activation via the P2Y12-Mediated Signaling Pathway

DOI：10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0406

摘要

Abstract

关键词

Keywords

references

CFTR Regulates Platelet Activation via the P2Y₁₂-Mediated Signaling Pathway