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山奈酚通过诱导自噬对大鼠脑缺血再灌注损伤的 AMPK/mTOR 信号通路具有保护作用。

Kaempferol Mediated AMPK/mTOR Signal Pathway Has a Protective Effect on Cerebral Ischemic-Reperfusion Injury in Rats by Inducing Autophagy.

机构信息

Department of Anesthesia, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, Tianjin, 300052, People's Republic of China.

Department of Pathology, Gansu Medical College, Lanzhou, 730050, Gansu, People's Republic of China.

出版信息

Neurochem Res. 2022 Aug;47(8):2187-2197. doi: 10.1007/s11064-022-03604-1. Epub 2022 May 7.

Abstract

Ischemia/reperfusion (I/R) caused by ischemic stroke treatments leads to brain injury and its pathological mechanism is related to autophagy. The underlying mechanism of kaempferol on cerebral I/R injury needs to be explored. To establish I/R injury, we used a middle cerebral artery occlusion-reperfusion (MCAO) model in rats. MCAO rats were treated with the same amount of saline (I/R group); Treatment group rats were treated orally with kaempferol (50, 100, 200 mg/kg) for 7 days before surgery. After reperfusion for 24 h, the scores of neurological deficits and infarct volume in each group were evaluated. LC3, Beclin-1 p62, AMPK and mTOR protein expression levels were examined by TTC staining, immunofluorescence staining, qRT-PCR and western blotting assay. H&E and TTC staining showed that compared with model group, the infarction size of rats in kaempferol group was markedly reduced. Meanwhile, the results showed that kaempferol had a dose-dependent nerve function repairability. Nissl and TUNEL staining showed that kaempferol could reduce neuronal apoptosis and ameliorate neuronal impairment after I/R. Western blotting and qRT-PCR results showed that kaempferol could protect the brain from ischemia reperfusion by activating autophagy. In addition, add AMPK inhibitor, western blotting and immumohistochemical staining showed that kaempferol mediated AMPK/mTOR signal pathway in MCAO rats. Kaempferol could mediate the AMPK signal pathway to regulate autophagy and inhibit apoptosis to protect brain against I/R injury.

摘要

缺血/再灌注(I/R)由缺血性中风治疗引起,导致脑损伤,其病理机制与自噬有关。山奈酚对脑 I/R 损伤的潜在机制需要进一步研究。为了建立 I/R 损伤模型,我们使用了大鼠大脑中动脉闭塞-再灌注(MCAO)模型。MCAO 大鼠用等量生理盐水(I/R 组)处理;治疗组大鼠在手术前用山奈酚(50、100、200mg/kg)口服治疗 7 天。再灌注 24h 后,评估各组神经功能缺损评分和梗死体积。通过 TTC 染色、免疫荧光染色、qRT-PCR 和 Western blot 检测 LC3、Beclin-1 p62、AMPK 和 mTOR 蛋白表达水平。H&E 和 TTC 染色结果表明,与模型组相比,山奈酚组大鼠的梗死面积明显减小。同时,结果表明山奈酚具有剂量依赖性的神经功能修复能力。尼氏染色和 TUNEL 染色结果表明,山奈酚可减少 I/R 后神经元凋亡,改善神经元损伤。Western blot 和 qRT-PCR 结果表明,山奈酚通过激活自噬来保护大脑免受缺血再灌注损伤。此外,添加 AMPK 抑制剂后,Western blot 和免疫组化染色结果表明,山奈酚介导了 MCAO 大鼠的 AMPK/mTOR 信号通路。山奈酚可以通过调节 AMPK 信号通路来调节自噬,抑制细胞凋亡,从而保护大脑免受 I/R 损伤。

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