Investigation of brain damage mechanism in middle cerebral artery occlusion/reperfusion rats based on i-TRAQ quantitative proteomics

• Published in: Experimental brain research 2021-04, Vol.239 (4), p.1247-1260
• Main Authors: Ma, Quantao, Wang, Chunguo, Wang, Min, Li, Yaqi, Li, Pengfei, Wang, Jingkang, Cheng, Long, An, Yongcheng, Dai, Hongyu, Duan, Yuhui, Wang, Ting, Zhao, Baosheng
• Format: Article
• Language: English
• Subjects: AKT2 protein; Apoptosis; Arterial occlusions; Biomedical and Life Sciences; Biomedicine; Brain damage; Brain injury; Brain research; Cdc42 protein; Cerebral blood flow; Cerebral cortex; Cerebral infarction; Cerebral ischemia; Complications and side effects; Development and progression; DNA damage; Gene regulation; Health aspects; Inflammation; Ischemia; Liquid chromatography; Mass spectroscopy; Metabolic pathways; Neurology; Neurosciences; NF-κB protein; Oxidative phosphorylation; Oxidative stress; Phosphorylation; Physiological aspects; Potentiation; Protein expression; Protein-protein interactions; Proteins; Proteomics; Reperfusion; Research Article; Rodents; Signal transduction; Therapeutic targets; Transcription; Triphenyltetrazolium chloride
• ISSN: 0014-4819; 1432-1106
• DOI: 10.1007/s00221-021-06054-3

The objective of this study is to analyze the differential protein expression profile in cerebral cortex of rats with middle cerebral ischemia/reperfusion (MCAO/R), explore the brain damage mechanism of MCAO/R at protein level, and provide experimental foundation for searching specific marker proteins of MCAO/R. Rat model of MCAO/R was established by modified suture-occluded method, and the model was evaluated by the results of brain 2,3,5-triphenyltetrazolium chloride (TTC) and hematoxylin–eosin (HE) staining. Cerebral cortex of rats from sham-operated group (Sham) and MCAO/R groups was used for FASP enzymatic hydrolysis, i-TRAQ quantitative labeling, and reverse-phase liquid chromatography purification and separation. Orbitrap Q Exactive mass spectrometry was used for qualitative and quantitative analyses of total differential protein expression profiles. MCAO/R rats had obvious cerebral infarction lesions, and the relative surface area of cerebral infarction was significantly different compared with sham rats, suggesting that MCAO/R rat model was successfully prepared. There were 199 significant difference proteins (MCAO/R vs Sham, p   1.2), including 104 up-regulated proteins and 95 down-regulated proteins. Gene ontology (GO) enrichment analysis showed that the up-regulated proteins were mainly concentrated in the biological processes of positive regulation of NF-κB transcription and I-κB kinase-NF-κB, etc. Down-regulated proteins were mainly concentrated in long-term synaptic potentiation, cellular response to DNA damage stimulus, etc. KEGG pathway analysis showed that the pathway involved in differential proteins includes oxidative phosphorylation, metabolic pathway, and Ras signaling pathway. Network analysis of differential proteins showed that Alb, ndufb5, ndufs7, ApoB, Cdc42, Ndufa3, Igf1r, P4hb, Mbp, Gc, Nme1, Akt2, and other proteins may play an important role in regulating oxidative stress, apoptosis, and inflammatory response in MCAO/R. Quantitative proteomics based on i-TRAQ labeling reveals the effect of inflammation and apoptosis in brain damage mechanism of MCAO/R. Besides, this research provide some experimental foundation for search and determination of potential therapeutic targets of MCAO/R.