Effects of nitric oxide synthase inhibitor Nω-nitro-L-arginine methyl ester on the expressions of autophagy in the penumbra of rats following focal cerebral ischemia/reperfusion

doi:10.3969/j.issn.1006-7795.2018.03.009

Abstract

Abstract: Objective To explore the effects of nitric oxide synthase(NOS) inhibitor Nω-nitro-L-arginine methyl ester (L-NAME) on the production of nitric oxide (NO) and the expression of Autophagy-related proteins Beclin1 and LC3B in the brain tissues of rats with middle cerebral artery occlusion (MCAO)/reperfusion injury and to investigate the mechanism of neuroprotective effect induced by L-NAME.Methods Eighteen male Sprague Dawley rats were divided into 3 groups randomly:Sham group, MCAO group, and L-NAME group (n=6). MCAO was induced by suture method. The rats were underwent 90 min of right MCAO, and then reperfusion by withdrawing filament. Rectal temperature was monitored and kept in normal range during the operation. The rats were sacrificed and the brains were harvested at 24 h after reperfusion. The expressions of Beclin1 and LC3B were detected by immunofluorescent staining. And the spatial location of 3-nitrotyrosine (3-NT), the biomarker of NO-mediated protein damage, and Beclin1/LC3B were detected respectively by double immunofluorescence labeling.Results In Sham group, no 3-NT positive cells were observed. Little Beclin1 positive cells and LC3B positive cells distributed in the brain tissues. Compared with Sham group, the number of Beclin1 positive cells and LC3B positive cells increased significantly in the penumbra of MCAO group (P<0.05). Compared with MCAO group, the number of Beclin1 positive cells and LC3B positive cells decreased significantly in the penumbra of rats after received L-NAME on 24 h after reperfusion (P<0.05). Beclin1 and LC3B-positive immunoreactive cells were colocalized with 3-NT separately in the penumbra of ischemia/reperfusion rats.Conclusion L-NAME reduced the production of NO by inhibiting the activity of NOS, which could prevent neurons from oxidative stress injury directly. At the same time, L-NAME also inhibited the expression of autophagy, which might play an indirect role in neuroprotective effect.

Key words: cerebral ischemia, nitric oxide synthase, autophagy, middle cerebral artery occlusion, rat

CLC Number:

R743.32

Huang Yuyou, Fang Yalan, Shi Wenjuan, Liu Kejian, Zhao Yongmei. Effects of nitric oxide synthase inhibitor Nω-nitro-L-arginine methyl ester on the expressions of autophagy in the penumbra of rats following focal cerebral ischemia/reperfusion[J]. Journal of Capital Medical University, 2018, 39(3): 355-359.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2018.03.009

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2018/V39/I3/355

References

[1] Kalkonde Y V, Deshmukh M D, Sahane V, et al. Stroke is the leading cause of death in rural gadchiroli, india:a prospective community-based study[J]. Stroke, 2015, 46(7):1764-1768.
[2] Bar-Or D, Bar-Or R, Rael L T, et al. Oxidative stress in severe acute illness[J]. Redox Biol, 2015, 4:340-345.
[3] Chamorro Á, Dirnagl U, Urra X, et al. Neuroprotection in acute stroke:targeting excitotoxicity, oxidative and nitrosative stress, and inflammation[J]. Lancet Neurol, 2016, 15(8):869-881.
[4] 黄语悠, 房亚兰, 刘克建等. 一氧化氮及其衍生物在缺血性脑血管病中的作用[J]. 首都医科大学学报, 2017, 38(1):67-71.
[5] Zhao H, Tao Z, Wang R, et al. MicroRNA-23a-3p attenuates oxidative stress injury in a mouse model of focal cerebral ischemia-reperfusion[J]. Brain Res, 2014, 1592:65-72.
[6] Liu H, Li J, Zhao F, et al.Nitric oxide synthase in hypoxic or ischemic brain injury[J].Rev Neurosci, 2015,26(1):105-117.
[7] Yuan Z, Liu W, Liu B, et al. Normobarichyperoxia delays and attenuates early nitric oxide production in focal cerebral ischemic rats[J].Brain Res, 2010,1352:248-254.
[8] Yang H, Chen Y, Yu L, et al. Esculentoside A exerts anti-inflammatory activity in microglial cells[J]. Int Immunopharmacol, 2017, 51:148-157.
[9] Yin Y, Zhao Y, Han S, et al. Autophagy-ERK1/2-Involved Disinhibition of Hippocampal Neurons Contributes to the Pre-Synaptic Toxicity Induced by Aβ42 Exposure[J]. J Alzheimers Dis, 2017, 59(3):851-869.
[10] Haiying L, Jiang W, Haitao S, et al. Autophagy in hemorrhagic stroke:mechanisms and clinical implications[J]. Prog Neurobiol,2017, 4(2):1-19.
[11] Feng J, Chen X, Guan B, et al. Inhibition of peroxynitrite-induced mitophagy activation attenuates cerebral ischemia-reperfusion injury[J]. Mol Neurobiol, 2018,[Epub ahead of print].
[12] Coelho C H, Martins T F, Oliveira-Pelegrin G R, et al. Inhibition of neuronal nitric oxide synthase activity does not alter vasopressin secretion in septic rats[J]. Pituitary, 2017,20(3):333-339.
[13] Shin J K, Kang J W, Lee S M. Enhanced nitric oxide-mediated autophagy contributes to the hepatoprotective effects of ischemic preconditioning during ischemia and reperfusion[J]. Nitric Oxide, 2016, 58:10-19.
[14] Bu Q, Liu X, Zhu Y, et al. w007B protects brain against ischemia-reperfusion injury in rats through inhibiting inflammation, apoptosis and autophagy[J]. Brain Res, 2014, 1558:100-108.
[15] Zhang X, Yan H, Yuan Y, et al. Cerebral ischemia-reperfusion-induced autophagy protects against neuronal injury by mitochondrial clearance[J]. Autophagy, 2013, 9(9):1321-1333.
[16] Ye L, Feng Z, Doycheva D, et al. CpG-ODN exerts a neuroprotective effect via the TLR9/pAMPK signaling pathway by activation of autophagy in a neonatal HIE rat model[J]. Exp Neurol, 2018, 301(Pt A):70-80.
[17] Xue T F, Ding X, Ji J, et al. PD149163 induces hypothermia to protect against brain injury in acute cerebral ischemic rats[J]. J Pharmacol Sci, 2017, 135(3):105-113.
[18] 曾晓莉, 张文娟,赵博,等.慢性脑缺血神经保护机制及药物研究进展[J].中国脑血管病杂志,2017,14(11):607-611.