Anti‐hypoxia effect of adenovirus‐mediated expression of heat shock protein 70 (HSP70) on primary cultured neurons. Issue 9 (17th May 2013)

• Record Type: Journal Article
• Authors: Hu, Dan; Chen, Fuqiang; Guan, Chun; Yang, Fangfang; Qu, Yan
• Is Part Of: Journal of neuroscience research. Volume 91:Issue 9(2013:Sep.)
• Page Start: 1174
• Page End: 1182
• Publication Date: 2013-05-17
• Subjects: Neurobiology -- Periodicals; 612
• DOI: 10.1002/jnr.23240
• Languages: English
• ISSNs: 0360-4012

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Heat shock protein 70 (HSP70) has attracted great attention recently in hypoxia injury because of its close link to the recovery after hypoxic–ischemic damage in organs. However, the cellular mechanism underlying its protective roles remains unclear. In this study, we developed a recombinant adenovirus containing HSP70‐GFP (vAd‐HSP70‐GFP) and studied the effect of virus‐mediated expression of exogenous HSP70 gene on neurons in response to hypoxia–reoxygenation injury. Virus‐mediated expression of HSP70 was detected as early as 24 hr and lasted until 10 days after infection. Neurons with 48 hr vAd‐HSP70‐GFP infection were exposed to 0, 0.5, 1, 2, 3, or 4 hr hypoxia followed by 1 hr reoxygenation. The mRNA and protein levels of HSP70 in neurons exposed to different lengths of hypoxia were compared by using RT‐PCR and Western blotting (WB). The 1‐hr hypoxia exposure showed the most significant increases in the HSP70 mRNA and protein level compared with other exposure durations. MTT assay showed that HSP70 overexpression significantly increased the neuronal viability, accompanied by decreased lactate dehydrogenase (LDH) activity in the culture medium after hypoxia–reoxygenation. Neurons with vAd‐HSP70‐GFP exhibited increased levels of mitochondrial cytochrome C (Cyt‐C) and decreased levels of cytoplasmic Cyt‐C compared with vAd‐GFP‐infected cells. These results suggest a neuroprotective role of exogenous HSP70 against hypoxia–reoxygenation injury, possibly via preventing initiation of mitochondrial apoptosis. © 2013 Wiley Periodicals, Inc.</p> </abstract>