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Antisense oligodeoxynucleotides of glial glutamate transporter--1 inhibits the neuro--protection of cerebral ischemic preconditioning in rats

Geng Jinxia, Cai Jingsong, Zhang Min, Li Shuqin, Sun Xiaocai, Xian Xiaohui, Hu Yuyan, Li Wenbin, Li Qingjun

Department of Pathophysiology,Hebei Medical University.Shijiazhuang 050017,Hebei

Abstract

The present study was undertaken to investigate the role of glial glutamate transporter-1 (GLT-1) in the brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP) by observing the effect of GLT-1 antisense oligodeoxynucleotides (AS-ODNs) on the neuro-protection of CIP against brain ischemic insult in rats. Wistar rats with permanently occluded bilateral vertebral arteries were randomly assigned to 7 groups: (1) Sham group: the bilateral common carotid arteries (BCCA) were separated, but without occluding the blood flow; (2) CIP group: the BCCA were clamped for 3 min; (3) Brain ischemic insult group: the BCCA were clamped for 8 min;(4) CIP + brain ischemic insult group: 3 min CIP was preformed 2 d prior to 8 min ischemic insult; (5) Double distilled water group: 5 #mu#L double distilled water was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively; (6) AS-ODNs group: 5 #mu#L AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively. This group was further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs; (7) AS-ODNs + CIP + brain ischemic insult group: 5 #mu#L AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after CIP, respectively. This group was also further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs. The other treatments were the same as those in CIP + brain ischemic insult group. The effect of the AS-ODNs on the expression of GLT-1 was assayed by using Western blot analysis. The profile of delayed neuronal death (DND) of pyramidal neurons in the CA1 hippocampus was evaluated by using thionin staining under light microscope by determining the neuronal density (ND) and histological grade (HG). Western blot analysis showed that AS-ODNs injected into the lateral cerebroventricle inhibited the expression of GLT-1 in the CA1 hippocampus in a dose-dependent manner. Neuropathological evaluation showed that there was no apparent DND in sham and CIP groups. Obvious DND of pyramidal neurons was found in brain ischemic insult group, which was represented by an increase in HG and a decrease in ND. CIP effectively protected the pyramidal neurons in the CA1 hippocampus against DND normally induced by ischemic insult, which indicating that CIP induced ischemic tolerance on the pyramidal neurons in the CA1 hippocampus. However, the injection of AS-ODNs into the lateral cerebroventricle blocked the neuro-protection of CIP against DND induced by brain ischemic insult. These results further proved the role of GLT-1 in the brain ischemic tolerance induced by CIP in rats.

Key words: cerebral ischemic preconditioning;brain ischemic tolerance;Hippocampus;glial glutamate transporter-1;antisense oligodeoxynucleotides;Rats

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Citing This Article：

Geng Jinxia, Cai Jingsong, Zhang Min, Li Shuqin, Sun Xiaocai, Xian Xiaohui, Hu Yuyan, Li Wenbin, Li Qingjun. Antisense oligodeoxynucleotides of glial glutamate transporter--1 inhibits the neuro--protection of cerebral ischemic preconditioning in rats. Acta Physiol Sin 2008; 60 (4): 497-503 (in Chinese with English abstract).