Sphingosine-1-phosphate type 1 receptor aggravates neointimal hyperplasia after carotid artery injury in mice

Hongqiang REN; Yijun LIU; Guiquan LUO; Li ZHAO

doi:10.16016/j.1000-5404.202009032

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Sphingosine-1-phosphate type 1 receptor aggravates neointimal hyperplasia after carotid artery injury in mice

Basic Medicine | 更新时间：2026-01-09

- Sphingosine-1-phosphate type 1 receptor aggravates neointimal hyperplasia after carotid artery injury in mice
- Journal of Third Military Medical University Vol. 43, Issue 3, Pages: 202-210(2021)
- 作者机构：
  
  629000 四川遂宁，遂宁市中心医院心血管内科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81770379)
- DOI：10.16016/j.1000-5404.202009032
  CLC： R363.21;R364.31;R543.4
- Received：05 September 2020，
  
  Revised：2020-10-05，
  
  Published：15 February 2021
- 稿件说明：
移动端阅览
Hongqiang REN, Yijun LIU, Guiquan LUO, et al. Sphingosine-1-phosphate type 1 receptor aggravates neointimal hyperplasia after carotid artery injury in mice[J]. Journal of Third Military Medical University, 2021, 43(3): 202-210.
DOI：

Hongqiang REN, Yijun LIU, Guiquan LUO, et al. Sphingosine-1-phosphate type 1 receptor aggravates neointimal hyperplasia after carotid artery injury in mice[J]. Journal of Third Military Medical University, 2021, 43(3): 202-210. DOI： 10.16016/j.1000-5404.202009032.

摘要

目的

探究1-磷酸鞘氨醇1型受体(sphingosine-1-phosphate type 1 receptor，S1PR1)对小鼠颈动脉损伤后血管内膜增生的作用及其机制。

方法

40只C57BL/6小鼠按随机数字表法分为4组:假手术+AAV-LacZ组、假手术+AAV-S1PR1组、导丝损伤+AAV-LacZ组和导丝损伤+AAV-S1PR1组，每组10只。各组小鼠尾静脉分别注射对应AAV-LacZ病毒或AAV-S1PR1病毒，2周后，假手术组仅切开颈部皮肤，颈动脉未损伤；导丝损伤组行颈动脉导丝损伤。4周后取颈动脉，HE染色检测新生内膜增生程度，组织免疫荧光检测颈动脉α-平滑肌肌动蛋白(α-smooth muscle actin，α-SMA)表达水平，组织免疫组化检测颈动脉组织增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)和IL-6表达水平。分离并培养小鼠原代主动脉血管平滑肌细胞，腺病毒Ad-S1PR1或Ad-LacZ转染后，加入1 μmol/L 1-磷酸鞘氨醇(sphingosine-1-phosphate，S1P)干预，Western blot和细胞免疫荧光检测原代主动脉平滑肌细胞表型转化，EdU染色检测细胞增殖，划痕实验和Transwell实验检测细胞迁移，Western blot检测IL-6/STAT3通路激活情况。

结果

小鼠颈动脉损伤4周后，与假手术+AAV-LacZ组相比，导丝损伤+AAV-LacZ组小鼠颈动脉可见显著的血管内膜增生、表型转化和细胞增殖(

0.05)；同时，导丝损伤后颈动脉IL-6表达上调。与导丝损伤+AAV-LacZ组相比，导丝损伤+AAV-S1PR1组颈动脉组织S1PR1过表达(

0.05)，上述病理生理学改变加重(

0.05)。细胞实验结果显示:S1P诱导血管平滑肌细胞表型转化和增殖迁移，激活IL-6/STAT3通路(

0.05)。腺病毒介导S1PR1过表达后，上述病理改变进一步加重(

0.05)。

结论

S1P/S1PR1促进血管平滑肌细胞表型转化和增殖迁移，加重血管损伤后内膜增生，可能与S1P/S1PR1激活血管平滑肌细胞IL-6/STAT3通路有关。

Abstract

Objective

To investigate the role and mechanism of sphingosine-1-phosphate type 1 receptor (S1PR1) in neointimal hyperplasia after carotid artery injury in mice.

Methods

A total of 40 C57BL/6 mice were rando

mly and equally subjected to 4 groups: sham operation+AAV-LacZ group

sham operation+AAV-S1PR1 group

model+AAV-LacZ group

and model+AAV-S1PR1 group. In 2 weeks after the delivery of AAV-LacZ or AAV-S1PR1 adenovirus to corresponding mice

via

tail vein injection

a mouse model of carotid artery injury was established with a flexible wire inserting into the common carotid artery. After 4 weeks

the carotid artery was harvested to measure the severity of neointimal hyperplasia by HE staining

the expression of α-smooth muscle actin (α-SMA) by immunofluorescence staining

and the levels of proliferating cell nuclear antigen (PCNA) and interleukin-6 (IL-6) by immunohistochemistry. In the isolated and primarily cultured mouse aortic vascular smooth muscle cells (VSMCs)

the adenovirus Ad-S1PR1 or Ad-LacZ were transfected respectively

and then the cells were incubated with 1 μmol/L S1P. The hallmarks of phenotypic switch of VSMCs were detected with Western blotting and immunofluorescence assay

cell proliferation with EdU staining

migration by scratch wound healing assay and transwell migration assay

and activation of IL-6/STAT3 pathway by Western blotting.

Results

In 4 weeks after model establishment

the mice of the model+AAV-LacZ group had obvious neointimal hyperplasia

phenotypic switch and cell proliferation (

0.05)

and significant upregulation of IL-6 in the carotid artery injury when compared with the mice of the sham operation+AAV-LacZ group. In the mice of the model+AAV-S1PR1 group

the level of S1PR1 was over-expressed (

0.05)

and above phenomena were aggravated when compared with the mice from the model+AAV-LacZ group (

0.05). In the

in vitro

study

S1P treatment induced the phenotypic switch

proliferation and migration

and activation of the IL-6/ STAT3 pathway in the primary VSMCs (

0.05)

and the pathophysiological changes above were further ag

gravated after the transfection of Ad-S1PR1 (

0.05).

Conclusion

S1P/S1PR1 can promote the phenotypic switch and proliferation and migration of VSMCs

and aggravate the intimal hyperplasia after vascular injury

which may be due to the activation of IL-6/STAT3 pathway.

关键词

Keywords

references

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