[脑室内注射腺苷A1受体反义寡脱氧核苷酸抑制大鼠脑缺血预处理的神经保护作用]

[Intracerebroventricular administration of adenosine A1 receptor antisense oligodeoxynucleotide inhibites the neuroprotective effect of the cerebral ischemic preconditioning in rats].

作者信息

Yun Xiao-jing, Hu Yu-yan, Xian Xiao-hui, Li Shu-qin, Sun Xiao-cai, Zhang Min, Li Qing-jun, Li Wen-bin

机构信息

Department of Pathophysiology, Institute of Basic Medicine, Hebei Medical University, Shijiazhuang 050017, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2008 Nov;24(4):430-3.

PMID:21158144

Abstract

AIM

To further explore the role of adenosine A1 receptor in the neuroprotective effect of cerebral ischemic preconditioning, the present study was undertaken to observe the effect of inhibiting expression of adenosine Al receptor with adenosine A1 receptor antisense oligodeoxynucleotide (ARA1 As-ODN) on the neuroprotective effect of cerebral ischemic preconditioning against delayed neuronal death (DND) normally induced by lethal brain ischemia.

METHOD

The rat 4-vessel occlusion global cerebral ischemic model was used. Forty-eight male Wistar rats with permanent occlusion of the bilateral vertebral arteries were divided into 8 groups: Sham, CIP, brain ischemic insult, CIP + brain ischemic insult, Distilled water + CIP + brain ischemic insult, ARA1 As-ODN, ARA1 As-ODN +CIP, ARA1 As-ODN+ CIP + brain ischemic insult(two doses of 10 nmol/5 microl and 20 nmol/5 microl were used) groups. ARA1 As-ODN was dissolved in distilled water and injected into the right lateral cerebral ventricle. To illustrate the profile of DND, histological grade (HG) and neuronal density (ND) in the CA1 region of the hippocampus were examined 7 d after the sham operation or the last time of ischemia under thionin staining.

RESULTS

The HG and ND in CIP group were similar to those in sham group. Brain ischemic insult induced obvious DND as represented with the increase in HG and decrease in ND significantly (P < 0.05 vs. sham and CIP groups). In CIP + ischemic insult group,no obvious DND was observed,which indicated that CIP protected pyramidal neurons against the ischemic insult.While the administration of ARA1 As-ODN in ARA1 As-ODN + CIP + brain ischemic insult group caused obvious increase in HG and decrease in ND compared with CIP + brain ischemic insult group (P < 0.05) in a dose dependent manner,which indicated that the neuroprotective effect of CIP against DND of hippocampal pyramidal neurons normally induced by ischemic insult was inhibited by the administration of ARA1 As-ODN.

CONCLUSION

The results further demonstrate the association of up-regulation of adenosine A1 receptors with the induction of CIP-mediated BIT.

摘要

目的

为进一步探讨腺苷A1受体在脑缺血预处理神经保护作用中的作用，本研究旨在观察用腺苷A1受体反义寡脱氧核苷酸（ARA1 As-ODN）抑制腺苷A1受体表达对脑缺血预处理抗致死性脑缺血所致延迟性神经元死亡（DND）神经保护作用的影响。

方法

采用大鼠四动脉闭塞全脑缺血模型。将48只永久性闭塞双侧椎动脉的雄性Wistar大鼠分为8组：假手术组、脑缺血预处理组、脑缺血损伤组、脑缺血预处理+脑缺血损伤组、蒸馏水+脑缺血预处理+脑缺血损伤组、ARA1 As-ODN组、ARA1 As-ODN+脑缺血预处理组、ARA1 As-ODN+脑缺血预处理+脑缺血损伤组（使用10 nmol/5 μl和20 nmol/5 μl两种剂量）。将ARA1 As-ODN溶于蒸馏水中，注入右侧侧脑室。为阐明DND的情况，在假手术或最后一次缺血7天后，用硫堇染色检查海马CA1区的组织学分级（HG）和神经元密度（ND）。

结果

脑缺血预处理组的HG和ND与假手术组相似。脑缺血损伤导致明显的DND，表现为HG增加和ND显著降低（与假手术组和脑缺血预处理组相比，P<0.05）。在脑缺血预处理+缺血损伤组中，未观察到明显的DND，这表明脑缺血预处理可保护锥体神经元免受缺血损伤。而在ARA1 As-ODN+脑缺血预处理+脑缺血损伤组中，与脑缺血预处理+脑缺血损伤组相比，给予ARA1 As-ODN导致HG明显增加和ND降低（P<0.05），且呈剂量依赖性，这表明给予ARA1 As-ODN可抑制脑缺血预处理对缺血损伤所致海马锥体神经元DND的神经保护作用。