Abstract

Context: Hypoxia-inducible factor-1α (HIF-1α)-induced genes can improve blood circulation.

Objective: To investigate brain protective effect of recombinant adenovirus-mediated HIF-1α (AdHIF-1α) expression and its mechanism.

Materials and methods: Male SD rats were used to establish focal cerebral ischaemia-reperfusion (CIR) injury models and randomly divided into normal, sham, CIR, Ad and AdHIF-1α groups. Ad or AdHIF-1α (10⁸ pfu/10 µL) were administered into lateral ventricle of rats in Ad and AdHIF-1α groups. Modified neurological severity score (mNSS), brain water content (BWC) and cerebral infarct volumes (CIVs) were analyzed, and HE staining was performed using the brain tissues. Furthermore, the expression of caspase-3 and HSP90 was analyzed using qRT-PCR and Western blotting.

Results: Compared to CIR (mNSS, 8.52 ± 0.52; CIV, 0.22 ± 0.01) and Ad groups (mNSS, 8.83 ± 0.41; CIV, 0.22 ± 0.02), mNSS and CIV were significantly decreased in AdHIF-1α group (mNSS, 6.03 ± 0.61; CIV, 0.11 ± 0.01) at 72 h (p < 0.05). With prolonged reperfusion time (6 h to 72 h), BWC of all rats increased gradually, although the increase was markedly less in AdHIF-1α group (78.15 ± 0.16 to 87.01 ± 0.31) compared to that in CIR (78.77 ± 0.60 to 89.74 ± 0.34) and Ad groups (78.77 ± 0.35 to 89.71 ± 0.27) (p < 0.01). There were significantly greater pathological changes in the neurons in AdHIF-1α group at 72 h following CIR. Furthermore, expression of caspase-3 (p < 0.01) down-regulated and HSP90 up-regulated (p < 0.05) at mRNA and protein levels in AdHIF-1α group.

Discussion and conclusions: HIF‑1α gene therapy is neuroprotective towards the CIR rat model. HIF-1α may be a candidate gene for the treatment of ischaemic brain injury.

摘要

背景：缺氧诱导因子-1α（HIF-1α）诱导的基因可以改善血液循环。

目的：探讨重组腺病毒介导的HIF-1α（AdHIF-1α）表达的脑保护作用及其机制。

材料和方法：雄性SD大鼠用于建立局灶性脑缺血再灌注（CIR）损伤模型，并随机分为正常，假，CIR，Ad和AdHIF-1α组。将Ad或AdHIF-1α（10 ⁸ pfu / 10 µL）注入Ad和AdHIF-1α组的大鼠侧脑室。分析修改后的神经系统严重程度评分（mNSS），脑含水量（BWC）和脑梗死体积（CIVs），并使用脑组织进行HE染色。此外，使用qRT-PCR和Western印迹分析了caspase-3和HSP90的表达。

结果：与CIR（mNSS，8.52±0.52; CIV，0.22±0.01）和Ad组（mNSS，8.83±0.41; CIV，0.22±0.02）相比，AdHIF-1α组（mNSS，6.03）的mNSS和CIV显着降低在72 h时±0.61; CIV，0.11±0.01）（p  <0.05）。随着再灌注时间的延长（6 h至72 h），所有大鼠的BWC逐渐增加，尽管AdHIF-1α组的增加明显少于CIR（78.77±0.60至89.74，从78.15±0.16至87.01±0.31）。 ±0.34）和广告组（78.77±0.35至89.71±0.27）（p  <0.01）。CIR后72小时，AdHIF-1α组神经元的病理变化明显更大。此外，caspase-3（p  <0.01）的表达下调而HSP90的表达上调（p <0.05）在AdHIF-1α组的mRNA和蛋白水平上。

讨论和结论： HIF-1α基因治疗对CIR大鼠模型具有神经保护作用。HIF-1α可能是治疗缺血性脑损伤的候选基因。