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. 2021 Apr 28:12:666429.
doi: 10.3389/fphar.2021.666429. eCollection 2021.

A Quantitative Serum Proteomic Analysis Helps to Explore the Comprehensive Mechanism and Identify Serum Biomarkers of Shengmai Injection's Effect on Isoproterenol-Induced Myocardial Ischemia in Rats

Xiaoping Zhang  1 , Jie Zhang  2 , Xiangyu Ji  2 , Zhenzhen Wei  2 , Baoyue Ding  2 , Guoqiang Liu  2 , Xiaoqing Lv  2 , Yongxia Zheng  2 , Shuyu Zhan  2
Affiliations

A Quantitative Serum Proteomic Analysis Helps to Explore the Comprehensive Mechanism and Identify Serum Biomarkers of Shengmai Injection's Effect on Isoproterenol-Induced Myocardial Ischemia in Rats

Xiaoping Zhang et al. Front Pharmacol. .

Abstract

Shengmai injection (SMI), a traditional Chinese medicine formula with the nature of multicomponent and multi-target, has been widely used in clinic for treating cardiovascular diseases in China; however, its comprehensive mechanism of action remains unclear. In this study, a TMT-based quantitative serum proteomics was performed to explore SMI's global mechanism and help identify serum biomarkers of its effect on isoproterenol (ISO)-induced myocardial ischemia rats. The results of TMT-based proteomic analysis identified 227, 100, and 228 differentially expressed proteins (DEPs) for the model compared to the control group, SMI pretreatment + model compared to the model group, and SMI pretreatment + model compared to the control group, respectively. Based on bioinformatics analyses of gene ontology (GO), KEGG pathways, and the protein-protein interaction (PPI) networks for the DEPs, it is concluded that the comprehensive mechanism of SMI's effect on ISO-induced myocardial ischemia injury includes regulation of energy metabolism, reducing endothelial cell permeability, regulation of vessel and cardiac contractility, anti-inflammation, and prevention of cell apoptosis. Furthermore, 10 common DEPs were found, and six of them were regulated in model vs. control group, while back-regulated in SMI pretreatment + model vs. model group. Among them, three functional proteins of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Fas apoptotic inhibitory molecule 3 (FAIM3), and uncharacterized protein (M0R5J4), which were verified by the PRM analysis, might be the potential serum biomarkers on SMI's effects. Overall, this serum proteomics of SMI not only provides insights into the comprehensive mechanism underlying SMI's effects on ischemic heart disease but also helps identify serum biomarkers for directing SMI's cardioprotective effects.

Keywords: Fas apoptotic inhibitory molecule 3; Shengmai injection; TMT; glyceraldehyde-3-phosphate dehydrogenase; myocardial ischemia; serum biomarkers; serum proteomics.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Workflow for exploring the comprehensive mechanism and identifying serum biomarkers of SMI’s cardioprotective effects.
FIGURE 2
FIGURE 2
Cardioprotective effect of SMI on ISO-induced myocardial ischemia injury in rats. (A) Representative H&E images (200×) of myocardial histology analysis for control, model, SMI-high, and SMI-low group rats. (B) Serum LDH, CK-MB, and SOD levels in control, model, SMI-high, and SMI-low group rats. (Data were expressed as Mean ± SD (n = 10), ## p < 0.01 vs. control, * p < 0.05 vs. model).
FIGURE 3
FIGURE 3
Differentially expressed proteins (DEPs) profiles by TMT-based proteomic analysis. (A, B, C) Heat maps of the overall differential concentrations of proteins quantified in Mod: Con (A), SMI: Mod (B), and SMI: Con (C), respectively. (D) Venn diagram showing overlap of all DEPs among Mod: Con, SMI: Mod, and SMI: Con.
FIGURE 4
FIGURE 4
Gene ontology (GO) analysis of all differentially expressed proteins (DEPs). Proteins were classified according to biological process (BP), molecular function (MF), and cellular component (CC) by Blast2GO software (version 3.3.5).
FIGURE 5
FIGURE 5
Top 20 enriched GO terms of significantly differentially expressed proteins (DEPs) in SMI: Mod. p value (more lower more intense in red color) represents the enriched degree. Each number above the bar charts is the richFactor (richFactor ≤1), which is the ratio of the number of proteins that have been annotated in this category.
FIGURE 6
FIGURE 6
Top 20 enriched KEGG pathways in SMI: Mod. p value (more lower more intense in red color) represents the enriched degree. Each number above the bar charts is the richFactor (richFactor ≤1), which is the ratio of the number of proteins that have been annotated in this category.
FIGURE 7
FIGURE 7
Seven protein-protein interaction (PPI) networks in SMI: Mod. The yellow node represents the differentially expressed proteins (DEPs), and the blue node represents the nonchanged protein. The most complex network involves 30 DEPs involved and Actb (ProteinID-P60711) has the highest connectivity degree of 15.
FIGURE 8
FIGURE 8
Average protein relative expression folds of the three proteins (M0R5J4, M0R590, and Q5M871) in Mod: Con, SMI: Mod, and SMI: Con by TMT and PRM, respectively. n = 3 samples for each group (3 data points for each bar), each consisting of serum of three rats.
See this image and copyright information in PMC

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