缺氧诱导线粒体DNA释放后激活cGAS-STING通路促进肺动脉平滑肌细胞增殖

韦汉南; 陈德伟; 张二龙; 高钰琪

doi:10.16016/j.2097-0927.202107134

您当前的位置：

首页 >

文章列表页 >

缺氧诱导线粒体DNA释放后激活cGAS-STING通路促进肺动脉平滑肌细胞增殖

基础医学 | 更新时间：2026-01-09

- 缺氧诱导线粒体DNA释放后激活cGAS-STING通路促进肺动脉平滑肌细胞增殖
- Hypoxia-induced mitochondrial DNA release promotes proliferation of pulmonary arterial smooth muscle cells via activating cGAS-STING pathway
- 陆军军医大学学报 2022年44卷第2期页码：117-124
- 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：
  
  [ 高钰琪，E-mail：gaoy66@yahoo.com GAO Yuqi, E-mail: gaoy66@yahoo.com
- 基金信息：
  
  国家自然科学基金重点项目(81830062)
- DOI：10.16016/j.2097-0927.202107134
  中图分类号： R322.121;R329.28;R394.2
- 收稿：2021-07-21，
  
  修回：2021-10-05，
  
  纸质出版：2022-01-30
- 稿件说明：
移动端阅览
韦汉南, 陈德伟, 张二龙, 等. 缺氧诱导线粒体DNA释放后激活cGAS-STING通路促进肺动脉平滑肌细胞增殖[J]. 陆军军医大学学报, 2022,44(2):117-124.

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.
韦汉南, 陈德伟, 张二龙, 等. 缺氧诱导线粒体DNA释放后激活cGAS-STING通路促进肺动脉平滑肌细胞增殖[J]. 陆军军医大学学报, 2022,44(2):117-124. DOI： 10.16016/j.2097-0927.202107134.

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

THENAPPAN T, ORMISTON M L, RYAN J J, et al . Pulmonary arterial hypertension: pathogenesis and clinical management[J ] . BMJ, 2018, 360: j5492. DOI: 10.1136/bmj.j5492.

高钰琪, 高文祥, 袁志兵, 等. 低氧肺血管炎症反应是高原肺水肿和低氧性肺动脉高压形成的关键环节[J]. 中国病理生理杂志, 2015, 31(10):1903. DOI: 10.3969/j.issn.1000-4718.2015.10.343.

GAO Y Q, GAO W X, YUAN Z B, et al . Hypoxic pulmonary vascular inflammatory response is a key link in the formation of plateau pulmonary edema and hypoxic pulmonary hypertension[J ] . Chin J Pathophysiol, 2015, 31(10): 1903. DOI: 10.3969/j.issn.1000-4718.2015.10.343.

PI W F, GUO X J, SU L P, et al . BMP-2up-regulates PTEN expression and induces apoptosis of pulmonary artery smooth muscle cells under hypoxia[J ] . PLoS One, 2012, 7(5): e35283. DOI: 10.1371/journal.pone.0035283.

WANG Z Y, YANG K, ZHENG Q Y, et al . Divergent changes of p53 in pulmonary arterial endothelial and smooth muscle cells involved in the development of pulmonary hypertension[J ] . Am J Physiol Lung Cell Mol Physiol, 2019, 316(1): L216-L228. DOI: 10.1152/ajplung.00538.2017.

GUI D, CUI Z, ZHANG L, et al . Salidroside attenuates hypoxia-induced pulmonary arterial smooth muscle cell proliferation and apoptosis resistance by upregulating autophagy through the AMPK-mTOR-ULK1 pathway[J ] . BMC Pulm Med, 2017, 17(1): 191. DOI: 10.1186/s12890-017-0477-4.

DROMPARIS P, MICHELAKIS E D. Mitochondria in vascular health and disease[J]. Annu Rev Physiol, 2013, 75: 95-126. DOI: 10.1146/annurev-physiol-030212-183804.

DASGUPTA A, WU D C, TIAN L, et al . Mitochondria in the pulmonary vasculature in health and disease: oxygen-sensing, metabolism, and dynamics[J ] . Compr Physiol, 2020, 10(2): 713-765. DOI: 10.1002/cphy.c190027.

COLLINS L V, HAJIZADEH S, HOLME E, et al . Endogenously oxidized mitochondrial DNA induces in vivo and in vitro inflammatory responses[J ] . J Leukoc Biol, 2004, 75(6): 995-1000. DOI: 10.1189/jlb.0703328..

MAEKAWA H, INOUE T, OUCHI H, et al . Mitochondrial damage causes inflammation via cGAS-STING signaling in acute kidney injury[J ] . Cell Rep, 2019, 29(5): 1261-1273. e6. DOI: 10.1016/j.celrep.2019.09.050..

OKA T, HIKOSO S, YAMAGUCHI O, et al . Mitochondrial DNA that escapes from autophagy causes inflammation and heart failure[J ] . Nature, 2012, 485(7397): 251-255. DOI: 10.1038/nature10992.

CAI X, CHIU Y H, CHEN Z J. The cGAS-cGAMP-STING pathway of cytosolic DNA sensing and signaling[J]. Mol Cell, 2014, 54(2): 289-296. DOI: 10.1016/j.molcel.2014.03.040.

KOVACS L, CAO Y P, HAN W H, et al . PFKFB 3 in smooth muscle promotes vascular remodeling in pulmonary arterial hypertension[J ] . Am J Respir Crit Care Med, 2019, 200(5): 617-627. DOI: 10.1164/rccm.201812-2290OC..

CHEN M X, DING Z Y, ZHANG F Y, et al . A20 attenuates hypoxia-induced pulmonary arterial hypertension by inhibiting NF-κB activation and pulmonary artery smooth muscle cell proliferation[J ] . Exp Cell Res, 2020, 390(2): 111982. DOI: 10.1016/j.yexcr.2020.111982.

KWON M Y, HWANG N, PARK Y J, et al . NOD2 deficiency exacerbates hypoxia-induced pulmonary hypertension and enhances pulmonary vascular smooth muscle cell proliferation[J ] . Oncotarget, 2018, 9(16): 12671-12681. DOI: 10.18632/oncotarget.23912.

WU D, DASGUPTA A, READ A D, et al . Oxygen sensing, mitochondrial biology and experimental therapeutics for pulmonary hypertension and cancer[J ] . Free Radic Biol Med, 2021, 170: 150-178. DOI: 10.1016/j.freeradbiomed.2020.12.452.

WU G N, ZHU Q Q, ZENG J L, et al . Extracellular mitochondrial D NA promote NLRP3 inflammasome activation and induce acute lung injury through TLR9 and NF-κB[J ] . J Thorac Dis, 2019, 11(11): 4816-4828. DOI: 10.21037/jtd.2019.10.26.

RIBEIRO JUNIOR R F, DABKOWSKI E R, SHEKAR K C, et al . MitoQ improves mitochondrial dysfunction in heart failure induced by pressure overload[J ] . Free Radic Biol Med, 2018, 117: 18-29. DOI: 10.1016/j.freeradbiomed.2018.01.012.

BI X J, DU C H, WANG X M, et al . Mitochondrial damage- induced innate immune activation in vascular smooth muscle cells promotes chronic kidney disease-associated plaque vulnerability[J ] . Adv Sci (Weinh), 2021, 8(5): 2002738. DOI: 10.1002/advs.202002738.

GUI X, YANG H, LI T, et al . Autophagy induction via STING trafficking is a primordial function of the cGAS pathway[J ] . Nature, 2019, 567(7747): 262-266. DOI: 10.1038/s41586-019-1006-9..

JING X G, JIANG T C, DAI L L, et al . Hypoxia-induced autophagy activation through NF-κB pathway regulates cell proliferation and migration to induce pulmonary vascular remodeling[J ] . Exp Cell Res, 2018, 368(2): 174-183. DOI: 10.1016/j.yexcr.2018.04.026.

SAKUMA M, TOYODA S, INOUE T, et al . Inflammation in pulmonary artery hypertension[J ] . Vasc Pharmacol, 2019, 118/119: 106562. DOI: 10.1016/j.vph.2019.05.002.

YOU N, LI J, HUANG X, et al . COMMD7 activates CXCL10 production by regulating NF-κB and the production of reactive oxygen species[J ] . Mol Med Rep, 2018, 17(5): 6784-6788. DOI: 10.3892/mmr.2018.8706.

NOZAKI E, KOBAYASHI T, OHNISHI H, et al . C-X-C motif receptor 3A enhances proliferation and invasiveness of colorectal cancer cells, and is mediated by C-X-C motif ligand 10[J ] . Oncol Lett, 2020, 19(3): 2495-2501. DOI: 10.3892/ol.2020.11326.

WU X M, SUN A Y, YU W F, et al . CXCL10 mediates breast cancer tamoxifen resistance and promotes estrogen-dependent and independent proliferation[J ] . Mol Cell Endocrinol, 2020, 512: 110866. DOI: 10.1016/j.mce.2020.110866.

FANG J X, WANG C, SHEN C, et al . The expression of CXCL10/CXCR3 and effect of the axis on the function of T lymphocyte involved in oral lichen planus[J ] . Inflammation, 2019, 42(3): 799-810. DOI: 10.1007/s10753-018-0934-0.

WANG X K, YUE T L, OHLSTEIN E H, et al . Interferon-inducible protein-10 involves vascular smooth muscle cell migration, proliferation, and inflammatory response[J ] . J Biol Chem, 1996, 271(39): 24286-24293. DOI: 10.1074/jbc.271.39.24286.

WANG H J, ZHOU Y, LIU R M, et al . IP-10/CXCR3 axis promotes the proliferation of vascular smooth muscle cells through ERK1/2/CREB signaling pathway[J ] . Cell Biochem Biophys, 2017, 75(1): 139-147. DOI: 10.1007/s12013-017-0782-9.

XIA J B, LIU G H, CHEN Z Y, et al . Hypoxia/ischemia promotes CXCL10 expression in cardiac microvascular endothelial cells by NFκB activation[J ] . Cytokine, 2016, 81: 63-70. DOI: 10.1016/j.cyto.2016.02.007.

GEORGE PM, OLIVER E, DORFMULLER P, et al . Evidence for the involvement of type I interferon in pulmonary arterial hypertension[J ] . Circ Res, 2014, 114(4): 677-688. DOI: 10.1161/CIRCRESAHA.114.302221.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

缺氧通过TAZ-H2A.Z轴调控H2A.Z染色质沉积促进肺泡上皮细胞增殖的研究

DNase 2α在缺氧诱导大鼠肺动脉平滑肌细胞增殖中的作用及机制研究

SOX6-SOCS3通过STAT3信号通路调控缺氧诱导的人肺动脉平滑肌细胞增殖

缺氧通过驱动M1巨噬细胞极化诱发睾丸间质细胞溶酶体依赖性死亡导致生精功能受损

MiR-616在骨肉瘤中的表达及对其细胞增殖、凋亡、迁移侵袭的作用