Hypoxia-induced mitochondrial DNA release promotes proliferation of pulmonary arterial smooth muscle cells <italic style="font-style: italic">via</italic> activating cGAS-STING pathway

Hannan WEI; Dewei CHEN; Erlong ZHANG; Yuqi GAO

doi:10.16016/j.2097-0927.202107134

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Hypoxia-induced mitochondrial DNA release promotes proliferation of pulmonary arterial smooth muscle cells via activating cGAS-STING pathway

Basic Medicine | 更新时间：2026-01-09

- Hypoxia-induced mitochondrial DNA release promotes proliferation of pulmonary arterial smooth muscle cells via activating cGAS-STING pathway
- Journal of Army Medical University Vol. 44, Issue 2, Pages: 117-124(2022)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系高原特需药品与器材研究室
  2.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系极端环境医学教育部重点实验室
  3.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系：病理生理学教研室
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(81830062)
- DOI：10.16016/j.2097-0927.202107134
  CLC： R322.121;R329.28;R394.2
- Received：21 July 2021，
  
  Revised：2021-10-05，
  
  Published：30 January 2022
- 稿件说明：
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Hannan WEI, Dewei CHEN, Erlong ZHANG, et al. Hypoxia-induced mitochondrial DNA release promotes proliferation of pulmonary arterial smooth muscle cells via activating cGAS-STING pathway[J]. Journal of Army Medical University, 2022, 44(2): 117-124.
DOI：

Hannan WEI, Dewei CHEN, Erlong ZHANG, et al. Hypoxia-induced mitochondrial DNA release promotes proliferation of pulmonary arterial smooth muscle cells via activating cGAS-STING pathway[J]. Journal of Army Medical University, 2022, 44(2): 117-124. DOI： 10.16016/j.2097-0927.202107134.

摘要

目的

探究线粒体DNA(mitochondrial DNA，mtDNA)释放在缺氧诱导大鼠肺动脉平滑肌细胞(rat pulmonary arterial smooth muscle cells，RPASMCs)增殖中的作用及可能机制。

方法

① 1%O

缺氧处理RPASMCs分3组(

=3)：常氧组、缺氧24 h组、缺氧48 h组。②环孢素A(CsA)抑制线粒体通透性转换孔(mitochondrial permeability transition pore，MPTP)分为3组(

=3)：常氧组、缺氧48 h组、缺氧48 h+CsA处理组。③siRNA敲低干扰素基因刺激因子(stimulator of interferon genes，STING)表达分为3组(

=3)：常氧+NC组、缺氧48 h+NC组、缺氧48 h+si-STING组。④采用CCK-8检测细胞增殖，RT-qPCR、Western blot检测环状GMP-AMP合酶(cyclic GMP-AMP synthase，cGAS)、STING、增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)的基因与蛋白表达，qPCR检测细胞质mtDNA释放，ELISA检测细胞培养上清中CXC趋化因子配体10(C-X-C motif chemokine ligand 10，CXCL10)水平。

结果

缺氧可诱导RPASMCs释放mtDNA(

0.01)，显著上调cGAS、STING、PCNA的表达(

0.05)，并增强CXCL10的分泌(

0.01)，进而促进RPASMCs增殖(0.44±0.02

0.72±0.03)(

0.05)。抑制MPTP可阻断mtDNA释放(

0.01)，显著下调cGAS、STING、PCNA的表达(

0.01)，降低CXCL10的分泌水平(

0.01)，并显著抑制缺氧诱导的RPASMCs增殖(0.74±0.12

0.43±0.06)(

0.01)。敲低STING可显著下调PCNA的表达(

0.05)，降低CXCL10的分泌水平(

0.01)，并显著抑制缺氧诱导的RPASMCs增殖(0.68±0.03

0.58±0.01)(

0.01)。

结论

缺氧可诱导平滑肌细胞释放mtDNA，介导缺氧诱导的平滑肌细胞增殖，其机制可能与cGAS-STING通路的激活有关。

Abstract

Objective

To investigate the role and possible mechanism of mitochondrial DNA (mtDNA) release in the proliferation of rat pulmonary arterial smooth muscle cells (RPASMCs) induced by hypoxia.

Methods

① RPASMCs were divided into normoxia group

hypoxia 24-hour group and hypoxia 48-hour group (

=3 for each groups)

and the latter 2 groups were exposed to 1%O

for 24 or 48 h respectively. ② Cyclosporine A (CsA) was used to inhibit the mitochondrial permeability transition pore (MPTP)

and then the RPASMCs were divided into normoxia

simple hypoxia 48-hour and hypoxia 48-hour+CsA groups (

=3 for each). ③ SiRNA was adopted to knockdown the expression of stimulator of interferon genes (STING)

and the cells were subsequently divided into normoxia+NC

hypoxia 48-hour+NC and hypoxia 48-hour+si-STING (

=3 for each). ④ The proliferation of the cells was measured by CCK-8 assay. The expression of cyclic GMP-AMP synthase (cGAS)

STING and proliferating cell nuclear antigen (PCNA) at mRNA and protein levels was detected by RT-qPCR and Western blotting

respectively. Cytosolic mtDNA release was determined by qPCR

and the level of C-X-C motif chemokine ligand 10 (CXCL10) in the culture supernatant was tested by ELISA.

Results

Hypoxia induced RPASMCs to release mtDNA (

0.01)

significantly up-regulated the expression of cGAS

STING and PCNA (

0.05)

enhanced the secretion of CXCL10 (

0.01)

and thus promoted the proliferation of RPASMCs (0.44±0.02

0.72±0.03

0.05). However

inhibition of MPTP blocked mtDNA release (

0.01)

down-regulated cGAS

STING and PCNA (

0.01)

and decreased CXCL10 level (

0.01) and hypoxia-induced proliferation of RPASMCs (0.74±0.12

0.43±0.06

0.01). The knockdown of STING also suppressed PCNA expression (

0.05)

reduced CXCL10 secretion (

0.01)

and inhibited the proliferation of RPASMCs (0.68±0.03

0.58±0.01

0.01).

Conclusion

Hypoxia can induce smooth muscle cells to release mtDNA and promote the proliferation of RPASMCs

which may be related to the activation of the cGAS-STING pathway.

关键词

Keywords

references

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⁰

Hypoxia-induced mitochondrial DNA release promotes proliferation of pulmonary arterial smooth muscle cells via activating cGAS-STING pathway

DOI：10.16016/j.2097-0927.202107134

摘要

Abstract

关键词

Keywords

references