基于TMT和PRM技术筛选车叶草苷干预CUMS大鼠海马组织的差异表达蛋白

doi:10.16431/j.cnki.1671-7236.2023.11.020

Abstract

Abstract: 【Objective】 This study was aimed to explore the protective mechanism of asperuloside on hippocampus in chronic unpredictable mild stress (CUMS) rats based on proteomics, and provide a basis for the development and utilization of asperuloside.【Method】 Forty-five male SD rats were chosen and randomly divided into three groups after one week pre-feed with 15 rats in each group, including the blank, model and asperuloside groups.In the blank group, 3 rats were raised in one cage, while in the other groups, one cage for one rat.The experimental period was 5 weeks, stimulated mice while medication administration starting from the 3rd week of the experiment.The rats in blank and model groups were given equal volume of saline by oral administration while in the asperuloside group was given a dose of 0.070 g/kg BW by intravenous injection once a day.The total hippocampus protein was extracted and the proteomics analysis of the hippocampus of depression rats was carried out by tandem mass tags (TMT)-LC-MS/MS technique to screen potential differential expression proteins.In addition, parallel reaction monitoring (PRM) was used to verify the expression of selected biomarkers in hippocampus.【Result】 A total of 6 up-regulated and 17 down-regulated proteins were screened between asperuloside and model groups.GO function and KEGG pathway enrichment analysis showed that the differentially expressed proteins were involved in signaling pathways such as neuroactive ligand-receptor interaction, complementary and synergistic regulation cascade and gastric acid secretion.PRM was used to verify the expression trend of bradykinin, immunoglobulin (IGHM and ILD) and proteasome activator complex subunit 1 (PSME1) were consistent with TMT quantitative results, it might be potential target proteins for the regulation of depression by asperuloside.Asperuloside could protect hippocampus through various pathways such as controlling key targets involved in regulating neuroactive ligand receptor interactions signaling pathways, exerting direct or indirect protective effect on neurons by down-regulating BK protein expression, up-regulating PSME1 protein levels significantly correlated with IgG leads to immunosuppression and normalization of immune protein concentration.【Conclusion】 Asperuloside might alleviate depression symptoms through multiple synergistic effects.The results could provide a references for further exploring the pathological mechanisms of the CUMS model and the proteins or pathways regulated by asperuloside in hippocampus.

Key words: tandem mass tags (TMT); parallel reaction monitoring (PRM); asperuloside; proteomics; hippocampus

CLC Number:

S816.41

YIN Li, LU Chengshu, ZENG Guofang, ZENG Shiyuan, LYU Jiayu, WANG Huakun. Screening of Differentially Expressed Proteins in Asperuloside Intervention CUMS Rats Hippocampus Based on TMT and PRM Techniques[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4513-4525.

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Screening of Differentially Expressed Proteins in Asperuloside Intervention CUMS Rats Hippocampus Based on TMT and PRM Techniques

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