Proteomics Analysis of Mechanism of Shenxiong Glucose Injection in Antagonizing H2O2-Induced Oxidative Damage in H9c2 Cells
10.13422/j.cnki.syfjx.20211417
- VernacularTitle:基于蛋白质组学分析参芎葡萄糖注射液拮抗H2O2诱导H9c2细胞氧化损伤的作用机制
- Author:
Ding-yan LU
1
;
Zhong-xiu WU
2
;
Jia SUN
2
;
Yuan LU
2
;
Yong-lin WANG
2
;
Yong-jun LI
1
;
Lin ZHENG
2
;
Ting LIU
2
Author Information
1. Engineering Research Center for the Development and Application of Ethnic Medicine and Traditional Chinese Medicine，Ministry of Education，State Key Laboratory of Functions and Applications of Medicinal Plants，Guizhou Medical University，Guiyang 550004，China
2. Provincial Key Laboratory of Pharmaceutics in Guizhou Province，Guizhou Medical University， Guiyang 550004，China
- Publication Type:Journal Article
- Keywords:
Shenxiong glucose injection （SGI）;
oxidative damage;
proteomics;
peroxisome proliferator-activated receptor （PPAR） pathway;
focal adhesion pathway;
phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）pathway;
Ras-related protein 1 （Rap1） pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(1):141-149
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the mechanism of Shenxiong glucose injection （SGI） in inhibiting hydrogen peroxide （H2O2）-induced oxidative damage in H9c2 cells by tandem mass tags （TMT）-labeled quantitative proteomics. MethodH9c2 cells cultured in vitro were exposed to H2O2 for inducing oxidative damage. The cell viability was measured by cell proliferation and cytotoxicity assay （MTS）， followed by peptide fractionation by high performance liquid chromatography （HPLC） and protein expression detection in H9c2 cells by ultrahigh performance liquid chromatography-mass spectrometry. MaxQuant （v1.5.2.8） was utilized for data retrieval， and the high-resolution mass spectrometry was conducted to screen out differentially expressed proteins， which were then subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis. The protein expression levels of perilipin 2 （Plin2） and tropomyosin 1 （Tpm1） in cells were measured by Western blot. ResultThe spectral analysis yielded 48 608 specific peptide fragments and 5 903 quantifiable proteins. Compared with the model group，the SGI group exhibited 82 differentially expressed proteins，of which 22 were up-regulated and 60 were down-regulated. GO analysis results showed that the differentially expressed proteins were mainly involved in biological processes such as programmed cell death regulation，regulation of cell proliferation，cardiovascular system development， and cell migration. As revealed by KEGG analysis， these proteins were mainly related to peroxisome proliferator-activated receptor （PPAR），focal adhesion，phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt），and Ras-related protein 1 （Rap1） pathways. Western blot results demonstrated that compared with the model group，SGI significantly increased the Plin2 protein expression and decreased the Tpm1 protein expression （P<0.01），consistent with the proteomics results. ConclusionSGI may inhibit cell apoptosis and antagonize H2O2-induced cell oxidative damage by regulating PPAR，focal adhesion，PI3K/Akt and Rap1 pathways，which should be further verified by subsequent experiments.
