机构:[1]Department of Neurology, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou, China广东省中医院[2]State Key Laboratory of Dampness Syndrome of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China广东省中医院[3]Department of Second Institute of Clinical Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou, China[4]Guangdong Provincial Key Laboratory of Research on Emergency in TCM, Guangzhou, China[5]Department of Anatomy, Sun Yat-Sen School of Medicine, Sun Yat-Sen University, Shenzhen, China深圳市康宁医院深圳医学信息中心中国医学科学院阜外医院深圳医院[6]Faculty of Medicine, Macau University of Science and Technology, Macau, China
Background: Oxidative stress plays an important role in the pathology of ischemic stroke. Studies have confirmedthat scutellarin has antioxidant effects against ischemic injury, and we also reported that the involvement of Aldose reductase (AR) in oxidative stress and cerebral ischemic injury, in this study we furtherly explicit whether the antioxidant effect of scutellarin on cerebral ischemia injury is related to AR gene regulation and its specific mechanism. Methods: C57BL/6N mice (Wild-type, WT) and AR knockout (AR(-/-)) mice suffered from transient middle cerebral artery occlusion (tMCAO) injury (1 h occlusion followed by 3 days reperfusion), and scutellarin was administered from 2 h before surgery to 3 days after surgery. Subsequently, neurological function was assessed by the modified Longa score method, the histopathological morphology observed with 2,3,5-triphenyltetrazolium chloride (TTC) and hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (Elisa) was used to detect the levels of ROS, 4-hydroxynonenal (4-HNE), 8-hydroxydeoxyguanosine (8-OHDG), Neurotrophin-3 (NT-3), poly ADP-ribose polymerase-1 (PARP1) and 3-nitrotyrosine (3-NT) in the ischemic penumbra regions. Quantitative proteomics profiling using quantitative nano-HPLC-MS/MS were performed to compare the protein expression difference between AR(-/- )and WT mice with or without tMCAO injury. The expression of AR, nicotinamide adenine dinucleotide phosphate oxidases (NOX1, NOX2 and NOX4) in the ipsilateral side of ischemic brain were detected by qRT-PCR, Western blot and immunofluorescence co-staining with NeuN. Results: Scutellarin treatment alleviated brain damage in tMCAO stroke model such as improved neurological function deficit, brain infarct area and neuronal injury and reduced the expression of oxidation-related products, moreover, also down-regulated tMCAO induced AR mRNA and protein expression. In addition, the therapeutic effect of scutellarin on the reduction of cerebral infarction area and neurological function deficits abolished in AR- /- miceunder ischemia cerebral injury, which indicated that the effect of scutellarin treatment on tMCAO injury is through regulating AR gene. Proteomic analysis of AR(-/-)and WT mice indicated AR knockout would affect oxidation reaction even as NADPH related process and activity in mice under cerebral ischemia conditions. Moreover, NOX isoforms (NOX1, NOX2 and NOX4) mRNA and protein expression were significant decreased in neurons of penumbra region in AR(-/- )mice compared with that in WT mice at 3d after tMCAO injury, which indicated that AR should be the upstream protein regulating NOX after cerebral ischemia. Conclusions: We first reported that AR directly regulates NOX subtypes (not only NOX2 but also NOX1 and NOX4) after cerebral ischaemic injury. Scutellarin specifically targets the AR-NOX axis and has antioxidant effects in mice with cerebral ischaemic injury, providing a theoretical basis and accurate molecular targets for the clinical application of scutellarin.
基金:
This study was financially supported by the National Natural Science Foundation of China, No. 81303115, 81774042, 81771353 and 81904104; the Pearl River S & T Nova Program of Guangzhou, No.201806010025; the Specialty Program of Guangdong Province Hospital of Traditional Chinese Medicine of China, No. YN2016MJ07, 2015KT1294, YN2015MS02, and YN2018ZD07 and the Youth Pilot Project of Chinese Society of Traditional Chinese Medicine, No. CACM-2018-QNRC2-C09; the Natural Science Foundation of Guangdong Province of China, No. S2013040016915; the Science and Technology Program of Guangzhou City of China, No. 202102010268; the Postdoctoral Foundation of China, No.2021M690759, BBK42913K09, 201003345, and BBH429151701; Hong Kong Scholar Program; Guangdong Provincial Key Laboratory of Research on Emergency in TCM, No.2017B030314176, 2018-75 and 2019-140; Guangzhou University of TCM 2017 high level University Construction Program, No. A1- AFD018171Z11096; the Administration of Traditional Chinese Medicine of Guangdong Province, No.20211203; and Research Fund for Zhaoyang Talents of Guangdong Provincial Hospital of Chinese Medicine, No. ZY2022KY06.
第一作者机构:[1]Department of Neurology, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou, China[3]Department of Second Institute of Clinical Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou, China
共同第一作者:
通讯作者:
通讯机构:[1]Department of Neurology, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou, China[2]State Key Laboratory of Dampness Syndrome of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China[3]Department of Second Institute of Clinical Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou, China[4]Guangdong Provincial Key Laboratory of Research on Emergency in TCM, Guangzhou, China[*1]Department of Neurology, Guangdong Provincial Hospital of Traditional Chinese Medicine, 111 Dade Road, Guangzhou, 510120, China.
推荐引用方式(GB/T 7714):
Deng Minzhen,Sun Jingbo,Peng Lilin,et al.Scutellarin acts on the AR-NOX axis to remediate oxidative stress injury in a mouse model of cerebral ischemia/reperfusion injury[J].PHYTOMEDICINE.2022,103:doi:10.1016/j.phymed.2022.154214.
APA:
Deng, Minzhen,Sun, Jingbo,Peng, Lilin,Huang, Yan,Jiang, Wen...&Cheng, Xiao.(2022).Scutellarin acts on the AR-NOX axis to remediate oxidative stress injury in a mouse model of cerebral ischemia/reperfusion injury.PHYTOMEDICINE,103,
MLA:
Deng, Minzhen,et al."Scutellarin acts on the AR-NOX axis to remediate oxidative stress injury in a mouse model of cerebral ischemia/reperfusion injury".PHYTOMEDICINE 103.(2022)