Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion <italic style="font-style: italic">via</italic> the ROS-RIP1/RIP3-exosome axis

Xue Zeng; Yun-Dong Zhang; Rui-Yan Ma; Yuan-Jing Chen; Xin-Ming Xiang; Dong-Yao Hou; Xue-Han Li; He Huang; Tao Li; Chen-Yang Duan

doi:10.1186/s40779-022-00383-2

Your Location：

Home >

Browse articles >

Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis

RESEARCH | Updated：2023-03-20

- Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis
- Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis
- 解放军医学杂志（英文版） 2022年9卷第6期页码：668-685
- Affiliations：
  
  1.Department of Anaesthesiology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China
  2.Department of Neurology, the Third Affiliated Hospital of Chongqing Medical University, Chongqing 401120, China
  3.Department of Cardiovascular Surgery, Xinqiao Hospital, Army Medical University, Chongqing 400037, China
  4.State Key Laboratory of Trauma, Burns and Combined Injury, Department of Shock and Transfusion, Daping Hospital, Army Medical University, Chongqing 400042, China
- Author bio：
  
  * 13708385559@163.com;
  lt200132@126.com;
  duanchenyang1991@cqmu.edu.cn
- Funds：
  
  the National Natural Science Foundation of China(81700429);the China Postdoctoral Science Foundation(2021MD703924);the Chongqing Postdoctoral Innovative Talents Support Program(CQBX2021018);the Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University
- DOI：10.1186/s40779-022-00383-2
  中图分类号：
- 纸质出版：2022-12
- Accepted：
Scan QR Code
Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis[J]. 解放军医学杂志（英文版）, 2022,9(6):668-685.

Zeng X, Zhang YD, Ma RY, Chen YJ, Xiang XM, Hou DY, et al. Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis. Mil Med Res. 2022;9(1):25.
Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis[J]. 解放军医学杂志（英文版）, 2022,9(6):668-685. DOI： 10.1186/s40779-022-00383-2.

Zeng X, Zhang YD, Ma RY, Chen YJ, Xiang XM, Hou DY, et al. Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis. Mil Med Res. 2022;9(1):25. DOI： 10.1186/s40779-022-00383-2.

摘要

Abstract

Background:

Cerebral ischemia-reperfusion injury (CIRI) refers to a secondary brain injury that can occur when the blood supply to the ischemic brain tissue is restored. However

the mechanism underlying such injury remains elusive.

Methods:

The 150 male C57 mice underwent middle cerebral artery occlusion (MCAO) for 1 h and reperfusion for 24 h

Among them

50 MCAO mice were further treated with Mitochondrial division inhibitor 1 (Mdivi-1) and 50 MCAO mice were further treated with N-acetylcysteine (NAC). SH-SY5Y cells were cultured in a low-glucose culture medium for 4 h under hypoxic conditions and then transferred to normal conditions for 12 h. Then

cerebral blood flow

mitochondrial structure

mitochondrial DNA (mtDNA) copy number

intracellular and mitochondrial reactive oxygen species (ROS)

autophagic flux

aggresome and exosome expression profiles

cardiac tissue structure

mitochondrial length and cristae density

mtDNA and ROS content

as well as the expression of Drp1-Ser616/Drp1

RIP1/RIP3

LC3 II/I

TNF-α

IL-1β

etc.

were detected under normal or Drp1 interference conditions.

Results:

The mtDNA content

ROS levels

and Drp1-Ser616/Drp1 were elevated by 2.2

1.7 and 2.7 times after CIRI (

0.05). However

the high cytoplasmic LC3 II/I ratio and increased aggregation of p62 could be reversed by 44% and 88% by Drp1 short hairpin RNA (shRNA) (

0.05). The low fluorescence intensity of autophagic flux and the increased phosphorylation of RIP3 induced by CIRI could be attenuated by ROS scavenger

NAC (

0.05). RIP1/RIP3 inhibitor Necrostatin-1 (Nec-1) restored 75% to a low LC3 II/I ratio and enhanced 2 times to a high RFP-LC3 after Drp1 activation (

0.05). In addition

although CIRI-induced ROS production caused no considerable accumulation of autophagosomes (

0.05)

it increased the packaging and extracellular secretion of exosomes containing p62 by 4–5 times

which could be decreased by Mdivi-1

Drp1 shRNA

and Nec-1 (

0.05). Furthermore

TNF-α and IL-1β increased in CIRI-derived exosomes could increase RIP3 phosphorylation in normal or oxygen–glucose deprivation/reoxygenation (OGD/R) conditions (

0.05).

Conclusions:

CIRI activated Drp1 and accelerated the p62-mediated formation of autophagosomes while inhibiting the transition of autophagosomes to autolysosomes

via

the RIP1/RIP3 pathway activation. Undegraded autophagosomes were secreted extracellularly in the form of exosomes

leading to inflammatory cascades that further damaged mitochondria

resulting in excessive ROS generation and the blockage of autophagosome degradation

triggering a vicious cycle.

关键词

Keywords

references

Campbell BCV , De Silva DA , Macleod MR , Coutts SB , Schwamm LH , Davis SM , et al . Ischaemic stroke . Nat Rev Dis Primers . 2019 ; 5 ( 1 ): 70 .

Wu X , Lin L , Qin JJ , Wang L , Wang H , Zou Y , et al . CARD3 promotes cerebral ischemia-reperfusion injury via activation of TAK1 . J Am Heart Assoc . 2020 ; 9 ( 9 ): e014920 .

Gong L , Tang Y , An R , Lin M , Chen L , Du J . RTN1-C mediates cerebral ischemia/reperfusion injury via ER stress and mitochondria-associated apoptosis pathways . Cell Death Dis . 2017 ; 8 ( 10 ): e3080 .

Nakamura Y , Lo EH , Hayakawa K . Placental mitochondria therapy for cerebral ischemia-reperfusion injury in mice . Stroke . 2020 ; 51 ( 10 ): 3142 - 6 .

Song J , Herrmann JM , Becker T . Quality control of the mitochondrial proteome . Nat Rev Mol Cell Biol . 2021 ; 22 ( 1 ): 54 - 70 .

Anzell AR , Maizy R , Przyklenk K , Sanderson TH . Mitochondrial quality control and disease: insights into ischemia-reperfusion injury . Mol Neurobiol . 2018 ; 55 ( 3 ): 2547 - 64 .

Haileselassie B , Joshi AU , Minhas PS , Mukherjee R , Andreasson KI , Mochly-Rosen D . Mitochondrial dysfunction mediated through dynamin-related protein 1 (Drp1) propagates impairment in blood brain barrier in septic encephalopathy . J Neuroinflammation . 2020 ; 17 ( 1 ): 36 .

Duan C , Kuang L , Xiang X , Zhang J , Zhu Y , Wu Y , et al . Drp1 regulates mitochondrial dysfunction and dysregulated metabolism in ischemic injury via Clec16a-, BAX-, and GSH- pathways . Cell Death Dis . 2020 ; 11 ( 4 ): 251 .

Duan C , Cao Z , Tang F , Jian Z , Liang C , Liu H , et al . miRNA-mRNA crosstalk in myocardial ischemia induced by calcified aortic valve stenosis . Aging . 2019 ; 11 ( 2 ): 448 - 66 .

Duan C , Wang L , Zhang J , Xiang X , Wu Y , Zhang Z , et al . Mdivi-1 attenuates oxidative stress and exerts vascular protection in ischemic/hypoxic injury by a mechanism independent of Drp1 GTPase activity . Redox Biol . 2020 ; 37 : 101706 .

Duan C , Kuang L , Xiang X , Zhang J , Zhu Y , Wu Y , et al . Activated Drp1-mediated mitochondrial ROS influence the gut microbiome and intestinal barrier after hemorrhagic shock . Aging . 2020 ; 12 ( 2 ): 1397 - 416 .

Chen L , Chen XY , Wang QL , Yang SJ , Zhou H , Ding LS , et al . Astragaloside IV derivative (LS-102) alleviated myocardial ischemia reperfusion injury by inhibiting Drp1Ser616 phosphorylation-mediated mitochondrial fission . Front Pharmacol . 2020 ; 11 : 1083 .

Yu X , Jia L , Yu W , Du H . Dephosphorylation by calcineurin regulates translocation of dynamin-related protein 1 to mitochondria in hepatic ischemia reperfusion induced hippocampus injury in young mice . Brain Res . 2019 ; 1711 : 68 - 76 .

Pugsley HR . Assessing autophagic flux by measuring LC3, p62, and LAMP1 co-localization using multispectral imaging flow cytometry . J Vis Exp . 2017 ; 125 : 55637 .

Zhang M , Zhao Z , Shen M , Zhang Y , Duan J , Guo Y , et al . Polydatin protects cardiomyocytes against myocardial infarction injury by activating Sirt3 . Biochim Biophys Acta Mol Basis Dis . 2017 ; 1863 ( 8 ): 1962 - 72 .

Fan RZ , Guo M , Luo S , Cui M , Tieu K . Exosome release and neuropathology induced by alpha-synuclein: new insights into protective mechanisms of Drp1 inhibition . Acta Neuropathol Commun . 2019 ; 7 ( 1 ): 184 .

Wen M , Jin Y , Zhang H , Sun X , Kuai Y , Tan W . Proteomic analysis of rat cerebral cortex in the subacute to long-term phases of focal cerebral ischemia-reperfusion injury . J Proteome Res . 2019 ; 18 ( 8 ): 3099 - 118 .

Song Y , Li Z , He T , Qu M , Jiang L , Li W , et al . M2 microglia-derived exosomes protect the mouse brain from ischemia-reperfusion injury via exosomal miR-124 . Theranostics . 2019 ; 9 ( 10 ): 2910 - 23 .

Ni HM , Williams JA , Ding WX . Mitochondrial dynamics and mitochondrial quality control . Redox Biol . 2015 ; 4 : 6 - 13 .

Yoo SM , Jung YK . A Molecular approach to mitophagy and mitochondrial dynamics . Mol Cells . 2018 ; 41 ( 1 ): 18 - 26 .

Calabrese V , Cornelius C , Dinkova-Kostova AT , Calabrese EJ , Mattson MP . Cellular stress responses, the hormesis paradigm, and vitagenes: novel targets for therapeutic intervention in neurodegenerative disorders . Antioxid Redox Signal . 2010 ; 13 ( 11 ): 1763 - 811 .

Pennisi M , Crupi R , Di Paola R , Ontario ML , Bella R , Calabrese EJ , et al . Inflammasomes, hormesis, and antioxidants in neuroinflammation: role of NRLP3 in Alzheimer disease . J Neurosci Res . 2017 ; 95 ( 7 ): 1360 - 72 .

Zhang Y , He Y , Wu M , Chen H , Zhang L , Yang D , et al . Rehmapicroside ameliorates cerebral ischemia-reperfusion injury via attenuating peroxynitritemediated mitophagy activation . Free Radic Biol Med . 2020 ; 160 : 526 - 39 .

Lee DS , Kim JE . PDI-mediated S-nitrosylation of DRP1 facilitates DRP1-S616 phosphorylation and mitochondrial fission in CA1 neurons . Cell Death Dis . 2018 ; 9 ( 9 ): 869 .

Ikeda Y , Shirakabe A , Maejima Y , Zhai P , Sciarretta S , Toli J , et al . Endogenous Drp1 mediates mitochondrial autophagy and protects the heart against energy stress . Circ Res . 2015 ; 116 ( 2 ): 264 - 78 .

Shirakabe A , Zhai P , Ikeda Y , Saito T , Maejima Y , Hsu CP , et al . Drp1-dependent mitochondrial autophagy plays a protective role against pressure overload-induced mitochondrial dysfunction and heart failure . Circulation . 2016 ; 133 ( 13 ): 1249 - 63 .

Wu W , Lin C , Wu K , Jiang L , Wang X , Li W , et al . FUNDC1 regulates mitochondrial dynamics at the ER-mitochondrial contact site under hypoxic conditions . EMBO J . 2016 ; 35 ( 13 ): 1368 - 84 .

Yamada T , Murata D , Adachi Y , Itoh K , Kameoka S , Igarashi A , et al . Mitochondrial stasis reveals p62-mediated ubiquitination in parkin-independent mitophagy and mitigates nonalcoholic fatty liver disease . Cell Metab . 2018 ; 28 ( 4 ): 588 - 604 .

Yamada T , Dawson TM , Yanagawa T , Iijima M , Sesaki H . SQSTM1/p62 promotes mitochondrial ubiquitination independently of PINK1 and PRKN/parkin in mitophagy . Autophagy . 2019 ; 15 ( 11 ): 2012 - 8 .

Yamada T , Adachi Y , Yanagawa T , Iijima M , Sesaki H . p62/sequestosome-1 knockout delays neurodegeneration induced by Drp1 loss . Neurochem Int . 2018 ; 117 : 77 - 81 .

Lan R , Wu JT , Wu T , Ma YZ , Wang BQ , Zheng HZ , et al . Mitophagy is activated in brain damage induced by cerebral ischemia and reperfusion via the PINK1/Parkin/p62 signalling pathway . Brain Res Bull . 2018 ; 142 : 63 - 77 .

Wang X , Li L , Niu X , Dang X , Li P , Qu L , et al . mTOR enhances foam cell formation by suppressing the autophagy pathway . DNA Cell Biol . 2014 ; 33 ( 4 ): 198 - 204 .

Otsubo K , Maeyashiki C , Nibe Y , Tamura A , Aonuma E , Matsuda H , et al . Receptor-interacting protein kinase 3 (RIPK3) inhibits autophagic flux during necroptosis in intestinal epithelial cells . FEBS Lett . 2020 ; 594 ( 10 ): 1586 - 95 .

Zhou K , Shi L , Wang Z , Zhou J , Manaenko A , Reis C , et al . RIP1-RIP3-DRP1 pathway regulates NLRP3 inflammasome activation following subarachnoid hemorrhage . Exp Neurol . 2017 ; 295 : 116 - 24 .

Rayamajhi M , Miao EA . The RIP1-RIP3 complex initiates mitochondrial fission to fuel NLRP3 . Nat Immunol . 2014 ; 15 ( 12 ): 1100 - 2 .

Li C , Mu N , Gu C , Liu M , Yang Z , Yin Y , et al . Metformin mediates cardioprotection against aging-induced ischemic necroptosis . Aging Cell . 2020 ; 19 ( 2 ): e13096 .

Wu X , Showiheen SAA , Sun AR , Crawford R , Xiao Y , Mao X , et al . Exosomes extraction and identification . Methods Mol Biol . 2019 ; 2054 : 81 - 91 .

Yang F , Ning Z , Ma L , Liu W , Shao C , Shu Y , et al . Exosomal miRNAs and miRNA dysregulation in cancer-associated fibroblasts . Mol Cancer . 2017 ; 16 ( 1 ): 148 .

Zhang J , Li S , Li L , Li M , Guo C , Yao J , et al . Exosome and exosomal microRNA: trafficking, sorting, and function . Genom Proteom Bioinform . 2015 ; 13 ( 1 ): 17 - 24 .

Kalluri R , LeBleu VS . The biology, function, and biomedical applications of exosomes . Science . 2020 ; 367 ( 6478 ): eaau6977 .

Islam MN , Das SR , Emin MT , Wei M , Sun L , Westphalen K , et al . Mitochondrial transfer from bone-marrow-derived stromal cells to pulmonary alveoli protects against acute lung injury . Nat Med . 2012 ; 18 ( 5 ): 759 - 65 .

Phinney DG , Di Giuseppe M , Njah J , Sala E , Shiva S , St Croix CM , et al . Mesenchymal stem cells use extracellular vesicles to outsource mitophagy and shuttle microRNAs . Nat Commun . 2015 ; 6 : 8472 .

Babuta M , Furi I , Bala S , Bukong TN , Lowe P , Catalano D , et al . Dysregulated autophagy and lysosome function are linked to exosome production by micro-RNA 155 in alcoholic liver disease . Hepatology . 2019 ; 70 ( 6 ): 2123 - 41 .

Hessvik NP , Overbye A , Brech A , Torgersen ML , Jakobsen IS , Sandvig K , et al . PIKfyve inhibition increases exosome release and induces secretory autophagy . Cell Mol Life Sci . 2016 ; 73 ( 24 ): 4717 - 37 .

浏览量

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

工具集

关联资源

GSTM1 suppresses cardiac fibrosis post-myocardial infarction through inhibiting lipid peroxidation and ferroptosis

TGF-β1 mediates hypoxia-preconditioned olfactory mucosa mesenchymal stem cells improved neural functional recovery in Parkinson’s disease models and patients

ALDH2 in autophagy and cell death: molecular mechanisms and implications for diseases

Immune checkpoint inhibition mediated with liposomal nanomedicine for cancer therapy

开裂预应力混凝土梁的检测、评估和加固

Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis

Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis

DOI：10.1186/s40779-022-00383-2

摘要

Abstract

关键词

Keywords

references