Effects of nitric oxide on nerve growth factor and brain derived neurotrophic factor expressions in rats with focal cerebral ischemia/reperfusion injury

doi:10.3969/j.issn.1006-7795.2020.06.008

Abstract

Abstract: Objective To investigate the dynamic changes of nerve growth factor (NGF) and brain derived neurotrophic factor (BDNF) during reperfusion time in a rat middle cerebral artery occlusion (MCAO)/reperfusion model and to observe the effects of nitric oxide synthase (NOS) inhibitor Nω-nitro-L-argininic methyl ester (L-NAME) on the expression of NGF and BDNF, in order to explore the deep mechanism of cerebral ischemia/reperfusion injury. Methods A total of 42 male Sprague Dawley rats were divided randomly into 7 groups according to random number table: Sham group, MCAO with 0h reperfusion (MCAO 0 h) group, MCAO with 6h reperfusion (MCAO 6 h) group, MCAO with 12 h reperfusion (MCAO 12 h) group, MCAO with 24 h reperfusion (MCAO 24 h) group, MCAO with 72 h reperfusion (MCAO 72 h) group and MCAO 24 h+L-NAME group. L-NAME was administered at a dose of 1 mg/kg by intraperitoneal injection at 30 minutes before MCAO. N=6 for each group. MCAO operation was performed by using suture method, the rats underwent 90 min of right MCAO, and then were reperfused by withdrawal of the filament. The expression of NGF and BDNF in the ischemic penumbra of frozen sections of the brain tissues were detected by immunofluorescence staining. The colocalization of 3-nitrotyrosine (3-NT), the marker of NO-mediated protein damage, with NGF and BDNF, was detected by immunofluorescence double staining respectively. Results There were no NGF and BDNF positive cells in the penumbra of MCAO 0 h and MCAO 6 h groups, while few of NGF and BDNF positive cells were observed in the penumbra of MCAO 12 h group. Compared with the MCAO 12 h group, the expressions of NGF and BDNF in the ischemic penumbra of MCAO 24 h group increased significantly (P<0.05). However, the expressions of NGF and BDNF in the penumbra of MCAO 72 h group were significantly decreased in contrast with MCAO 24 h group (P<0.05). In the penumbra of brain tissue of MCAO 24 h group, 3-NT was colocalized with NGF and BDNF. There were no 3-NT and NGF double staining cell in the sham group. In the MCAO 24 h group, lots of 3-NT and NGF double staining positive cells were observed in the ischemic penumbra. After treated with L-NAME, the number of 3-NT and NGF double staining positive cells in the penumbra of the ischemia/reperfusion rats were significantly decreased compared with that of MCAO 24 h group (P<0.05). There were no 3-NT and BDNF double staining cells in the sham group. In the ischemic penumbra of MCAO 24 h group rats, amounts of 3-NT and BDNF double staining positive cells were observed. Compared with MCAO 24 h group,the numbers of 3-NT and BDNF double staining positive cells in the penumbra of the ischemia/reperfusion rats were significantly decreased after treatment with L-NAME (P<0.05). Conclusions Cerebral ischemia /reperfusion induced the upregulation of the expression of NGF and BDNF in the penumbra of ischemic rats. The initial upregulated expression appeared at about 12 h after reperfusion, the expression of NGF and BDNF increased with the prolongation of reperfusion time and reached peak at 24 h after reperfusion, followed with a significant decrease at 72 h after reperfusion. L-NAME reduced the production of NO by inhibiting the activity of NOS, leading to the reduction of 3-NT, and then reduce the expression of NGF and BDNF, indicating that NO induces the expression of NGF and BDNF during cerebral ischemia/reperfusion injury.

Key words: cerebral ischemia/reperfusion, nitric oxide, nerve growth factor, brain derived neurotrophic factor, rat

CLC Number:

R743.31

Yang Nan, Ding Mao, Huang Yuyou, Fang Yalan, Shi Wenjuan, Zhao Yongmei. Effects of nitric oxide on nerve growth factor and brain derived neurotrophic factor expressions in rats with focal cerebral ischemia/reperfusion injury[J]. Journal of Capital Medical University, 2020, 41(6): 908-913.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2020.06.008

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2020/V41/I6/908

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