Effect of autophagy inhibitor 3-MA on inflammatory response in the brain of rats with cerebral ischemia/reperfusion injury

doi:10.3969/j.issn.1006-7795.2024.06.010

Abstract

Abstract: Objective To investigate the effects of the autophagy inhibitor 3-methyladenine (3-MA) on C-X-C chemokine receptor type 2 (CXCR2), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and cyclooxygenase-2 (COX2) in the ischemic brain tissue of rats subjected to transient focal cerebral ischemia/reperfusion, in order to explore the role of autophagy in inflammation during cerebral ischemia/reperfusion injury. Methods The male Sprague Dawley rats were randomized into: sham group, middle cerebral artery occlusion (MCAO) group and MCAO+3-MA (3-MA) group, with n=5 for each group. A model of MCAO was induced by using the intraluminal suture method. The rats underwent 90 minutes of ischemia. Rats in 3-MA group were intraperitoneally injected with 10 mg/kg of 3-MA before MCAO. All groups of rats underwent reperfusion for 24 h. After reperfusion time, the rats were quickly decapitated, and their brains were removed. The levels of CXCR2 protein in the ischemic brain tissue were detected by Western blotting. The number of positive cells of TNF-α, IL-1β, and COX2 were detected by immunofluorescence staining in the ischemic penumbra, and the cellular localization of IL-1β and COX2 was evaluated respectively. Results ①Compared with sham group, the level of CXCR2 protein was upregulated obviously in the ischemic brain tissue of MCAO rats after 24 h reperfusion (P < 0.05). The level of CXCR2 protein decreased significantly in the ischemic brain tissue of 3-MA group compared with that of MCAO group (P < 0.05). ②There were few TNF-α, IL-1β, and COX2 positive cells in the sham group. Compared with sham group, the number of TNF-α, IL-1β, and COX2 positive cells in the ischemic penumbra of MCAO group rats obviously increased after 24 h of reperfusion (P < 0.05). Compared with MCAO group, the number of positive cells of TNF-α, IL-1β and COX2 decreased significantly in the ischemic penumbra of the MCAO+3-MA group rats (P < 0.05). ③The IL-1β and COX2 positive cells were colocalized with NeuN, a general neuronal marker, in the ischemic penumbra of cerebral ischemia/reperfusion rats. Conclusions The autophagy inhibitor 3-MA may attenuate inflammation by inhibiting the expression levels of chemokine receptor CXCR2, as well as the number of positive cells of TNF-α, IL-1β, and COX2 inflammatory factors in ischemic brain tissue of MCAO rats after reperfusion, indicating that autophagy is one of the factors that promote the inflammatory response after ischemia.

Key words: cerebral ischemia/reperfusion injury, inflammation, autophagy

CLC Number:

R743.3

An Qi, Yang Nan, Zhu Yuequan, Shi Wenjuan, Huang Minqi, Zhao Yongmei. Effect of autophagy inhibitor 3-MA on inflammatory response in the brain of rats with cerebral ischemia/reperfusion injury[J]. Journal of Capital Medical University, 2024, 45(6): 1008-1015.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2024.06.010

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2024/V45/I6/1008

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