In-Depth Proteomic Analysis of the Hippocampus in a Rat Model after Cerebral Ischaemic Injury and Repair by Danhong Injection (DHI)

Abstract

Stroke is the second most common cause of death worldwide. A systematic description and characterization of the strokes and the effects induced in the hippocampus have not been performed so far. Here, we analysed the protein expression in the hippocampus 24 h after cerebral ischaemic injury and repair. Drug intervention using Danhong injection (DHI), which has been reported to have good therapeutic effects in a clinical setting, was selected for our study of cerebral ischaemia repair in rat models. A larger proteome dataset and total 4091 unique proteins were confidently identified in three biological replicates by combining tissue extraction for rat hippocampus and LC-MS/MS analysis. A label-free approach was then used to quantify the differences among the four experimental groups (Naive, Sham, middle cerebral artery occlusion (MCAO) and MCAO + DHI groups) and showed that about 2500 proteins on average were quantified in each of the experiment group. Bioinformatics analysis revealed that in total 280 unique proteins identified above were differentially expressed (P < 0.05). By combining the subcellular localization, hierarchical clustering and pathway information with the results from injury and repair phase, 12 significant expressed proteins were chosen and verified with respect to their potential as candidates for cerebral ischaemic injury by Western blot. The primary three signalling pathways of the candidates related may be involved in molecular mechanisms related to cerebral ischaemic injury. In addition, a glycogen synthase kinase-3β (Gsk-3β) inhibitor of the candidates with the best corresponding expression trends between western blotting (WB) and label-free quantitative results were chosen for further validation. The results of Western blot analysis of protein expression and 2,3,5- chloride three phenyl tetrazole (TTC) staining of rat brains showed that DHI treatment and Gsk-3β inhibitor are both able to confer protection against ischaemic injury in rat MCAO model. The observations of the present study provide a novel understanding regarding the regulatory mechanism of cerebral ischaemic injury.

Transcriptomic Study Reveals Recovery of Impaired Astrocytes Contribute to Neuroprotective Effects of Danhong Injection Against Cerebral Ischemia/Reperfusion-Induced Injury

“…Importantly, we performed microarray assay and provided global transcriptional data, which enabled us to explore molecular mechanisms contributing to protective effects of DHI against cerebral I/R injury. Although bioinformatics approaches, including proteomic analysis ( Cui et al, 2017 ) and systematic survey for transcription factors (TFs) changes ( Wei et al, 2016 ) have been applied, ours is to focus on the transcriptomic analysis, and on a late time point of 14 days.…”

Proteomic Assessment of iTRAQ-Based NaoMaiTong in the Treatment of Ischemic Stroke in Rats

“…As we all know, few studies exposed the mechanism of NMT on ischemic stroke. In this study, we used iTRAQ labeling and 2D LC-MS/MS to identify DEPs and altered signaling pathways following ischemic stroke in MCAO rat models [19]. We first confirmed that NMT reduced neurological damage and infarct area compared to the MCAO model group (p< 0.05, n=6) [20].…”

Work Patterns of MamXY Proteins during Magnetosome Formation in Magnetospirillum gryphiswaldense MSR-1

“…represents the interaction between MamZ and Mms6 that needs to be proven further. trospray ion source (Thermo Fisher) for proteomic analysis (34). Chromatography columns were packed with Ultimate XB-C 18 3-m resin (Welch Materials, MD, USA).…”