AdipoRon Protects Against Secondary Brain Injury After Intracerebral Hemorrhage via Alleviating Mitochondrial Dysfunction: Possible Involvement of AdipoR1–AMPK–PGC1α Pathway

  • Jun Yu
  • Jingwei Zheng
  • Jianan Lu
  • Zeyu Sun
  • Zefeng WangEmail author
  • Jianmin ZhangEmail author
Original Paper


Intracerebral hemorrhage (ICH) is a stroke subtype that is associated with high mortality and disability rate. Mitochondria plays a crucial role in neuronal survival after ICH. This study first showed that activation of adiponectin receptor 1 (AdipoR1) by AdipoRon could attenuate mitochondrial dysfunction after ICH. In vivo, experimental ICH model was established by autologous blood injection in mice. AdipoRon was injected intraperitoneally (50 mg/kg). Immunofluorescence staining were performed to explicit the location of AdipoR1, AMP-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor-γ coactivator-1a (PGC1α). The PI staining was used to quantify neuronal survival. The expression of AdipoR1 and its downstream signaling molecules were detected by Western blotting. In vitro, 10 μM oxyhemoglobin (OxyHb) was used to induce the neuronal injury in SH-SY5Y cells. Annexin V-FITC/PI staining was used to detect the neuronal apoptosis and necrosis. Mitochondrial membrane potential (Δψm) was measured by a JC-1 kit and mitochondrial mass was quantified by mitochondrial fluorescent probe. In vivo, PI staining showed that the administration of AdipoRon could reduce neuronal death at 72 h after ICH in mice. AdipoRon treatment enhanced ATP levels and reduced ROS levels in perihematoma tissues, and increased the protein expression of AdipoR1, P-AMPK, PGC1α, NRF1 and TFAM. In vitro, the JC-1 staining and Mito-tracker™ Green showed that AdipoRon significantly alleviated OxyHb-induced collapse of Δψm and enhanced mitochondrial mass. Moreover, flow cytometry analysis indicated that the neurons treated with AdipoRon showed low necrotic and apoptotic rate. AdipoRon alleviates mitochondrial dysfunction after intracerebral hemorrhage via the AdipoR1–AMPK–PGC1α pathway.


Mitochondrial dysfunction ICH AdipoRon ROS 



Intracerebral hemorrhage


Secondary brain injury


Reactive oxygen species


AMP-activated protein kinase




Fetal bovine serum


3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide


Phosphate buffered saline


Polyvinylidene fluoride


Magnetic resonance imaging


Peroxisome proliferator-activated receptor-γ coactivator-1a


Mitochondrial membrane potential


Nuclear respiratory factor 1


Mitochondrial transcription factor A


Author contributions

JMZ and ZFW are the principal investigators. JWZ and JY contributed to the study design, performance and manuscript draft. JWZ and JNL analyzed the experimental data revised the manuscript and polish the language.


This study was supported by Basic Public Interests Research Plan of Zhejiang Province (GF18H090006) to Jun Yu; The Major Research and Development Project of Zhejiang Province (2017C03021) to Jianmin Zhang.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Ethics Approval

All animal experimental protocols were in compliance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and were approved by the Institutional Animal Care and Use Committee of Zhejiang University.


  1. 1.
    Keep RF, Hua Y, Xi G (2012) Intracerebral haemorrhage: mechanisms of injury and therapeutic targets. Lancet Neurol 11(8):720–731CrossRefGoogle Scholar
  2. 2.
    Aronowski J, Zhao X (2011) Molecular pathophysiology of cerebral hemorrhage: secondary brain injury. Stroke 42(6):1781–1786CrossRefGoogle Scholar
  3. 3.
    Qureshi AI, Mendelow AD, Hanley DF (2009) Intracerebral haemorrhage. Lancet 373(9675):1632–1644CrossRefGoogle Scholar
  4. 4.
    Prentice H, Modi JP, Wu JY (2015) Mechanisms of neuronal protection against excitotoxicity, endoplasmic reticulum stress, and mitochondrial dysfunction in stroke and neurodegenerative diseases. Oxid Med Cell s 2015:964518Google Scholar
  5. 5.
    Zheng J, Shi L, Liang F, Xu W, Li T, Gao L et al (2018) Sirt3 ameliorates oxidative stress and mitochondrial dysfunction after intracerebral hemorrhage in diabetic rats. Front Neurosci 12:414CrossRefGoogle Scholar
  6. 6.
    Brunswick AS, Hwang BY, Appelboom G, Hwang RY, Piazza MA, Connolly ES Jr (2012) Serum biomarkers of spontaneous intracerebral hemorrhage induced secondary brain injury. J Neurol Sci 321(1–2):1–10CrossRefGoogle Scholar
  7. 7.
    Kim-Han JS, Kopp SJ, Dugan LL, Diringer MN (2006) Perihematomal mitochondrial dysfunction after intracerebral hemorrhage. Stroke 37(10):2457–2462CrossRefGoogle Scholar
  8. 8.
    Okada-Iwabu M, Yamauchi T, Iwabu M, Honma T, Hamagami K, Matsuda K et al (2013) A small-molecule AdipoR agonist for type 2 diabetes and short life in obesity. Nature 503(7477):7493CrossRefGoogle Scholar
  9. 9.
    Thundyil J, Pavlovski D, Sobey CG, Arumugam TV (2012) Adiponectin receptor signalling in the brain. Br J Pharmacol 165(2):313–327CrossRefGoogle Scholar
  10. 10.
    Duan J, Yin Y, Cui J, Yan J, Zhu Y, Guan Y et al (2016) Chikusetsu saponin IVa ameliorates cerebral ischemia reperfusion injury in diabetic mice via adiponectin-mediated AMPK/GSK-3beta pathway in vivo and in vitro. Mol Neurobiol 53(1):728–743CrossRefGoogle Scholar
  11. 11.
    Song J, Kang SM, Kim E, Kim CH, Song HT, Lee JE (2015) Adiponectin receptor-mediated signaling ameliorates cerebral cell damage and regulates the neurogenesis of neural stem cells at high glucose concentrations: an in vivo and in vitro study. Cell Death Dis 6(6):e1844CrossRefGoogle Scholar
  12. 12.
    Jiang T, Yu JT, Zhu XC, Zhang QQ, Tan MS, Cao L et al (2015) Ischemic preconditioning provides neuroprotection by induction of AMP-activated protein kinase-dependent autophagy in a rat model of ischemic stroke. Mol Neurobiol 51(1):220–229CrossRefGoogle Scholar
  13. 13.
    Zhang BB, Zhou G, Li C (2009) AMPK: an emerging drug target for diabetes and the metabolic syndrome. Cell Metab 9(5):407–416CrossRefGoogle Scholar
  14. 14.
    Mukherjee P, Mulrooney TJ, Marsh J, Blair D, Chiles TC, Seyfried TN (2008) Differential effects of energy stress on AMPK phosphorylation and apoptosis in experimental brain tumor and normal brain. Mol Cancer 7:37CrossRefGoogle Scholar
  15. 15.
    Sheng B, Wang X, Su B, Lee HG, Casadesus G, Perry G et al (2012) Impaired mitochondrial biogenesis contributes to mitochondrial dysfunction in Alzheimer’s disease. J Neurochem 120(3):419–429CrossRefGoogle Scholar
  16. 16.
    You Y, Hou YH, Zhai X, Li ZY, Li LY, Zhao Y et al (2016) Protective effects of PGC-1 alpha via the mitochondrial pathway in rat brains after intracerebral hemorrhage. Brain Res 1646:34–43CrossRefGoogle Scholar
  17. 17.
    Zhang XF, Ren XQ, Zhang Q, Li ZY, Ma SP, Bao JT et al (2016) PGC-1 alpha/ERR alpha-Sirt3 pathway regulates DAergic neuronal death by directly deacetylating SOD2 and ATP synthase beta. Antioxid Redox Signal 24(6):312–328CrossRefGoogle Scholar
  18. 18.
    Yu LM, Gong B, Duan WX, Fan CX, Zhang J, Li Z et al (2017) Melatonin ameliorates myocardial ischemia/reperfusion injury in type 1 diabetic rats by preserving mitochondrial function: role of AMPK-PGC-1 alpha-SIRT3 signaling. Sci Rep 7:41337CrossRefGoogle Scholar
  19. 19.
    Wang YJ, Liang B, Lau WB, Du YH, Guo R, Yan ZY et al (2017) Restoring diabetes-induced autophagic flux arrest in ischemic/reperfused heart by ADIPOR (adiponectin receptor) activation involves both AMPK-dependent and AMPK-independent signaling. Autophagy 13(11):1855–1869CrossRefGoogle Scholar
  20. 20.
    Ni W, Mao S, Xi G, Keep RF, Hua Y (2016) Role of erythrocyte CD47 in intracerebral hematoma clearance. Stroke 47(2):505–511CrossRefGoogle Scholar
  21. 21.
    Zhou K, Zhong Q, Wang YC, Xiong XY, Meng ZY, Zhao T et al (2017) Regulatory T cells ameliorate intracerebral hemorrhage-induced inflammatory injury by modulating microglia/macrophage polarization through the IL-10/GSK3 beta/PTEN axis. J Cereb Blood Flow Metab 37(3):967–979CrossRefGoogle Scholar
  22. 22.
    Gao MH, Wang JJ, Wang WX, Liu JS, Wong CW (2011) Phosphatidylinositol 3-kinase affects mitochondrial function in part through inducing peroxisome proliferator-activated receptor gamma coactivator-1 beta expression. Br J Pharmacol 162(4):1000–1008CrossRefGoogle Scholar
  23. 23.
    Rodriguez C, Sobrino T, Agulla J, Bobo-Jimenez V, Ramos-Araque ME, Duarte JJ et al (2017) Neovascularization and functional recovery after intracerebral hemorrhage is conditioned by the Tp53 Arg72Pro single-nucleotide polymorphism. Cell Death Differ 24(1):144–154CrossRefGoogle Scholar
  24. 24.
    Beray-Berthat V, Delifer C, Besson VC, Girgis H, Coqueran B, Plotkine M et al (2010) Long-term histological and behavioural characterisation of a collagenase-induced model of intracerebral haemorrhage in rats. J Neurosci Methods 191(2):180–190CrossRefGoogle Scholar
  25. 25.
    Sun J, Wei ZZ, Gu X, Zhang JY, Zhang Y, Li J et al (2015) Intranasal delivery of hypoxia-preconditioned bone marrow-derived mesenchymal stem cells enhanced regenerative effects after intracerebral hemorrhagic stroke in mice. Exp Neurol 272:78–87CrossRefGoogle Scholar
  26. 26.
    Liu Z, Zhang RL, Li Y, Cui Y, Chopp M (2009) Remodeling of the corticospinal innervation and spontaneous behavioral recovery after ischemic stroke in adult mice. Stroke 40(7):2546–2551CrossRefGoogle Scholar
  27. 27.
    Galho AR, Cordeiro MF, Ribeiro SA, Marques MS, Antunes MF, Luz DC et al (2016) Protective role of free and quercetin-loaded nanoemulsion against damage induced by intracerebral haemorrhage in rats. Nanotechnology 27(17):175101CrossRefGoogle Scholar
  28. 28.
    Piantadosi CA, Suliman HB (2012) Redox regulation of mitochondrial biogenesis. Free Radic Biol Med 53(11):2043–2053CrossRefGoogle Scholar
  29. 29.
    Huang JL, Manaenko A, Ye ZH, Sun XJ, Hu Q (2016) Hypoxia therapy–a new hope for the treatment of mitochondrial dysfunctions. Med Gas Res 6(3):174–176CrossRefGoogle Scholar
  30. 30.
    Zhou Y, Wang S, Li Y, Yu S, Zhao Y (2017) SIRT1/PGC-1alpha signaling promotes mitochondrial functional recovery and reduces apoptosis after intracerebral hemorrhage in rats. Front Mol Neurosci 10:443CrossRefGoogle Scholar
  31. 31.
    Wang Z, Zhou F, Dou Y, Tian X, Liu C, Li H et al (2018) Melatonin alleviates intracerebral hemorrhage-induced secondary brain injury in rats via suppressing apoptosis, inflammation, oxidative stress, DNA damage, and mitochondria injury. Transl Stroke Res 9(1):74–91CrossRefGoogle Scholar
  32. 32.
    Zeng J, Chen Y, Ding R, Feng L, Fu Z, Yang S et al (2017) Isoliquiritigenin alleviates early brain injury after experimental intracerebral hemorrhage via suppressing ROS- and/or NF-kappaB-mediated NLRP3 inflammasome activation by promoting Nrf2 antioxidant pathway. J Neuroinflamm 14(1):119CrossRefGoogle Scholar
  33. 33.
    Eleftheriadis T, Pissas G, Liakopoulos V, Stefanidis I (2016) Cytochrome c as a potentially clinical useful marker of mitochondrial and cellular damage. Front Immunol 7:279CrossRefGoogle Scholar
  34. 34.
    Finkel T, Holbrook NJ (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408(6809):239–247CrossRefGoogle Scholar
  35. 35.
    Bause AS, Haigis MC (2013) SIRT3 regulation of mitochondrial oxidative stress. Exp Gerontol 48(7):634–639CrossRefGoogle Scholar
  36. 36.
    Handschin C, Spiegelman BM (2006) Peroxisome proliferator-activated receptor gamma coactivator 1 coactivators, energy homeostasis, and metabolism. Endocr Rev 27(7):728–735CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Neurosurgery, Second Affiliated Hospital, School of MedicineZhejiang UniversityHangzhouChina
  2. 2.Brain Research InstituteZhejiang UniversityHangzhouChina
  3. 3.Collaborative Innovation Center for Brain ScienceZhejiang UniversityHangzhouChina

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