Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Overexpression of adiponectin promotes focal angiogenesis in the mouse brain following middle cerebral artery occlusion

Abstract

Recent studies indicate that overexpression of adiponectin (APN) could attenuate ischemic brain injury. However, the mechanism of APN effect remains unclear. In this study, we investigated the cellular mechanisms of APN action during cerebral ischemia. Adult mice (n=120) received an intracerebral injection of adeno-associated viral vector carrying the APN gene (AAV-APN). The mice were subjected to a transient ispilateral middle cerebral artery occlusion (tMCAO) after 7-day AAV-APN gene transfer. Cortical atrophy volume, neurological function, microvessels counts, phospho-AMPK and downstream angiogenic factor vascular endothelial growth factor (VEGF) were examined. Overexpression of APN was observed in the mouse brain following AAV-APN gene transfer. Cortical atrophy volume was attenuated in the AAV-APN-transduced mice compared with the AAV-GFP and saline-treated mice (7.9±0.6%, 19.8±0.3% and 20.3±1.1%, respectively, P<0.05), with significant improvement in neurological function and an increased number of microvessels (199±5 vs 151±4 and 148±4 mm–2, P<0.01). Furthermore, the expression of phospho-AMPK and VEGF were increased in the AAV-APN-transduced compared with the control mice (P<0.01), whereas inhibiting phospho-AMPK, reducing VEGF expression and attenuating the effect of APN on brain atrophy and angiogenesis (P<0.01). APN overexpression attenuates ischemia-induced brain atrophy and has improvement in neurological function. The consequence is related to promotion of focal angiogenesis. The AMPK signaling pathway has an important role in upregulating angiogenic factor VEGF.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Gladstone DJ, Black SE, Hakim AM . Toward wisdom from failure: lessons from neuroprotective stroke trials and new therapeutic directions. Stroke 2002; 33: 2123–2136.

    Article  Google Scholar 

  2. The national institute of neurological disorders and stroke rt-pa stroke study group. Tissue plasminogen activator for acute ischemic stroke. N Engl J Med 1995; 333: 1581–1587.

    Article  Google Scholar 

  3. Hacke W, Kaste M, Bluhmki E, Brozman M, Davalos A, Guidetti D et al. Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med 2008; 359: 1317–1329.

    Article  CAS  Google Scholar 

  4. Krupinski J, Kaluza J, Kumar P, Wang M, Kumar S . Prognostic value of blood vessel density in ischaemic stroke. Lancet 1993; 342: 742.

    Article  CAS  Google Scholar 

  5. Cramer SC, Nelles G, Benson RR, Kaplan JD, Parker RA, Kwong KK et al. A functional mri study of subjects recovered from hemiparetic stroke. Stroke 1997; 28: 2518–2527.

    Article  CAS  Google Scholar 

  6. Hao Q, Su H, Palmer D, Sun B, Gao P, Yang GY et al. Bone marrow-derived cells contribute to vascular endothelial growth factor-induced angiogenesis in the adult mouse brain by supplying matrix metalloproteinase-9. Stroke 2011; 42: 453–458.

    Article  CAS  Google Scholar 

  7. Zhu W, Mao Y, Zhao Y, Zhou LF, Wang Y, Zhu JH et al. Transplantation of vascular endothelial growth factor-transfected neural stem cells into the rat brain provides neuroprotection after transient focal cerebral ischemia. Neurosurgery 2005; 57: 325–333; discussion 325–333.

    Article  Google Scholar 

  8. Zhu Y, Lee C, Shen F, Du R, Young WL, Yang GY . Angiopoietin-2 facilitates vascular endothelial growth factor-induced angiogenesis in the mature mouse brain. Stroke 2005; 36: 1533–1537.

    Article  CAS  Google Scholar 

  9. Wang Y, Kilic E, Kilic U, Weber B, Bassetti CL, Marti HH et al. Vegf overexpression induces post-ischaemic neuroprotection, but facilitates haemodynamic steal phenomena. Brain 2005; 128: 52–63.

    Article  Google Scholar 

  10. Sun Y, Jin K, Xie L, Childs J, Mao XO, Logvinova A et al. Vegf-induced neuroprotection, neurogenesis, and angiogenesis after focal cerebral ischemia. J Clin Invest 2003; 111: 1843–1851.

    Article  CAS  Google Scholar 

  11. Zhang R, Wang L, Zhang L, Chen J, Zhu Z, Zhang Z et al. Nitric oxide enhances angiogenesis via the synthesis of vascular endothelial growth factor and cgmp after stroke in the rat. Circ Res 2003; 92: 308–313.

    Article  CAS  Google Scholar 

  12. Ouchi N, Walsh K . Adiponectin as an anti-inflammatory factor. Clin Chim Acta 2007; 380: 24–30.

    Article  CAS  Google Scholar 

  13. Takemura Y, Walsh K, Ouchi N . Adiponectin and cardiovascular inflammatory responses. Curr Atheroscler Rep 2007; 9: 238–243.

    Article  CAS  Google Scholar 

  14. Tilg H, Moschen AR . Role of adiponectin and pbef/visfatin as regulators of inflammation: involvement in obesity-associated diseases. Clin Sci (Lond) 2008; 114: 275–288.

    Article  CAS  Google Scholar 

  15. Chen MP, Tsai JC, Chung FM, Yang SS, Hsing LL, Shin SJ et al. Hypoadiponectinemia is associated with ischemic cerebrovascular disease. Arterioscler Thromb Vasc Biol 2005; 25: 821–826.

    Article  CAS  Google Scholar 

  16. Stott DJ, Welsh P, Rumley A, Robertson M, Ford I, Sattar N et al. Adipocytokines and risk of stroke in older people: a nested case-control study. Int J Epidemiol 2009; 38: 253–261.

    Article  Google Scholar 

  17. Efstathiou SP, Tsioulos DI, Tsiakou AG, Gratsias YE, Pefanis AV, Mountokalakis TD . Plasma adiponectin levels and five-year survival after first-ever ischemic stroke. Stroke 2005; 36: 1915–1919.

    Article  CAS  Google Scholar 

  18. Bang OY, Saver JL, Ovbiagele B, Choi YJ, Yoon SR, Lee KH . Adiponectin levels in patients with intracranial atherosclerosis. Neurology 2007; 68: 1931–1937.

    Article  CAS  Google Scholar 

  19. Sasaki M, Kawano T, Saito T, Yuzawa M, Saito T, Ikoma A et al. Hypoadiponectinemia in patients with cerebral infarction: comparison with other atherosclerotic disorders. Am J Med Sci 2007; 333: 140–144.

    Article  Google Scholar 

  20. Nishimura M, Izumiya Y, Higuchi A, Shibata R, Qiu J, Kudo C et al. Adiponectin prevents cerebral ischemic injury through endothelial nitric oxide synthase dependent mechanisms. Circulation 2008; 117: 216–223.

    Article  CAS  Google Scholar 

  21. Chen B, Liao WQ, Xu N, Xu H, Wen JY, Yu CA et al. Adiponectin protects against cerebral ischemia-reperfusion injury through anti-inflammatory action. Brain Res 2009; 1273: 129–137.

    Article  CAS  Google Scholar 

  22. Kobayashi H, Ouchi N, Kihara S, Walsh K, Kumada M, Abe Y et al. Selective suppression of endothelial cell apoptosis by the high molecular weight form of adiponectin. Circ Res 2004; 94: e27–e31.

    Article  CAS  Google Scholar 

  23. Shibata R, Ouchi N, Kihara S, Sato K, Funahashi T, Walsh K . Adiponectin stimulates angiogenesis in response to tissue ischemia through stimulation of amp-activated protein kinase signaling. J Biol Chem 2004; 279: 28670–28674.

    Article  CAS  Google Scholar 

  24. Shimano M, Ouchi N, Shibata R, Ohashi K, Pimentel DR, Murohara T et al. Adiponectin deficiency exacerbates cardiac dysfunction following pressure overload through disruption of an ampk-dependent angiogenic response. J Mol Cell Cardiol 2010; 49: 210–220.

    Article  CAS  Google Scholar 

  25. Yang GY, Xu B, Hashimoto T, Huey M, Chaly Jr T, Wen R et al. Induction of focal angiogenesis through adenoviral vector mediated vascular endothelial cell growth factor gene transfer in the mature mouse brain. Angiogenesis 2003; 6: 151–158.

    Article  CAS  Google Scholar 

  26. Choi VW, McCarty DM, Samulski RJ . Aav hybrid serotypes: improved vectors for gene delivery. Curr Gene Ther 2005; 5: 299–310.

    Article  CAS  Google Scholar 

  27. Conway EM, Collen D, Carmeliet P . Molecular mechanisms of blood vessel growth. Cardiovasc Res 2001; 49: 507–521.

    Article  CAS  Google Scholar 

  28. Yancopoulos GD, Davis S, Gale NW, Rudge JS, Wiegand SJ, Holash J . Vascular-specific growth factors and blood vessel formation. Nature 2000; 407: 242–248.

    Article  CAS  Google Scholar 

  29. Rosenstein JM, Mani N, Silverman WF, Krum JM . Patterns of brain angiogenesis after vascular endothelial growth factor administration in vitro and in vivo. Proc Natl Acad Sci USA 1998; 95: 7086–7091.

    Article  CAS  Google Scholar 

  30. Nagata D, Mogi M, Walsh K . Amp-activated protein kinase (ampk) signaling in endothelial cells is essential for angiogenesis in response to hypoxic stress. J Biol Chem 2003; 278: 31000–31006.

    Article  CAS  Google Scholar 

  31. Ouchi N, Shibata R, Walsh K . Amp-activated protein kinase signaling stimulates vegf expression and angiogenesis in skeletal muscle. Circ Res 2005; 96: 838–846.

    Article  CAS  Google Scholar 

  32. Shen F, Fan Y, Su H, Zhu Y, Chen Y, Liu W et al. Adeno-associated viral vector-mediated hypoxia-regulated vegf gene transfer promotes angiogenesis following focal cerebral ischemia in mice. Gene Therapy 2008; 15: 30–39.

    Article  CAS  Google Scholar 

  33. Yang G, Chan PH, Chen J, Carlson E, Chen SF, Weinstein P et al. Human copper-zinc superoxide dismutase transgenic mice are highly resistant to reperfusion injury after focal cerebral ischemia. Stroke 1994; 25: 165–170.

    Article  Google Scholar 

  34. Leon H, Atkinson LL, Sawicka J, Strynadka K, Lopaschuk GD, Schulz R . Pyruvate prevents cardiac dysfunction and amp-activated protein kinase activation by hydrogen peroxide in isolated rat hearts. Can J Physiol Pharmacol 2004; 82: 409–416.

    Article  CAS  Google Scholar 

  35. Fan Y, Shen F, Frenzel T, Zhu W, Ye J, Liu J et al. Endothelial progenitor cell transplantation improves long-term stroke outcome in mice. Ann Neurol 2010; 67: 488–497.

    Article  CAS  Google Scholar 

  36. Zhang L, Schallert T, Zhang ZG, Jiang Q, Arniego P, Li Q et al. A test for detecting long-term sensorimotor dysfunction in the mouse after focal cerebral ischemia. J Neurosci Methods 2002; 117: 207–214.

    Article  Google Scholar 

  37. Shen F, Su H, Liu W, Kan YW, Young WL, Yang GY . Recombinant adeno-associated viral vector encoding human vegf165 induces neomicrovessel formation in the adult mouse brain. Front Biosci 2006; 11: 3190–3198.

    Article  CAS  Google Scholar 

  38. Cui X, Chopp M, Zacharek A, Zhang C, Roberts C, Chen J . Role of endothelial nitric oxide synthetase in arteriogenesis after stroke in mice. Neuroscience 2009; 159: 744–750.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by the 973 Program 2011CB504405 (GYY, YW), the National Natural Science Foundation of China Project #30973097 (GYY), the Young Scientists Fund of the National Natural Science Foundation of China Project # 81100633 (LHS) and a grant from Shanghai Municipal Health Bureau #2010090 (LHS). We thank Ms Karena Shun and Yan Wang for their editorial assistance and staffs of the Neuroscience and Neuroengineering Center for their collaborative support.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Y Zhao or G-Y Yang.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shen, L., Miao, J., Yuan, F. et al. Overexpression of adiponectin promotes focal angiogenesis in the mouse brain following middle cerebral artery occlusion. Gene Ther 20, 93–101 (2013). https://doi.org/10.1038/gt.2012.7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/gt.2012.7

Keywords

This article is cited by

Search

Quick links