Anti-hypoxia effect of adenovirus-mediated expression of heat shock protein 70 (HSP70) on primary cu

• 作 者: DanHu[1]FuqiangChen[1]ChunGuan[1]FangfangYang[2]YanQu[1]
• 机 构: Department of Intensive Care Unit, the Municipal Hospital of Medical College of Qingdao University,Q
• 出 处: 山东病理生理学会危重病医学专业委员会第八届学术大会暨山东省医师协会急救医学医师分会第三届学术大会
• 发表日期: 2012年期

　　Heat shock protein 70 (HSP70) has attracted great attention recently in hypoxia injury due to its close link with recovery after hypoxic-ischemic damage in organs,while the cellular mechanism remains unclear.In this study,we generated 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 h after infection and lasted until 10 days.Neurons with 48h vAd-HSP70 infection were exposed to hypoxia with durations of 0,0.5,1,2,3,or 4 hours,followed with 1 hour reoxygenation.RT-PCR and western blotting (WB) were performed to detect the mRNA and protein level of HSP70 in neurons with hypoxia-reoxygenation.HSP70 mRNA reached maximal level at 1h and started decreasing at 2h after hypoxia-reoxygenation.Protein level of HSP70 showed a similar pattern as that of mRNA level in response to hypoxia injury.MTT assay showed that HSP70 expression significantly increased neuronal viability and it is associated with decreased lactate dehydrogenase (LDH) activity in culture medium after hypoxia-reoxygenation.Neurons with vAd-HSP70-GFP exhibited increased level of mitochondrial eytochrome C (Cyt-C) and decreased level of cytoplasmic Cyt-C compared to vAd-GFP-infected cells.These results suggest a neuroprotective role of exogenous HSP70 against hypoxia-reoxygenation injury possibly via preventing initiation of mitochondrial apoptosis.