Proteomics analysis of coronary atherosclerotic heart disease with different Traditional Chinese Medicine syndrome types before and after percutaneous coronary intervention

doi:10.19852/j.cnki.jtcm.20240408.001

Abstract

Abstract:

OBJECTIVE: To investigate the underlying protein molecular mechanisms of "Qi stagnation and blood stasis syndrome" (QS) and "Qi deficiency and blood stasis syndrome" (QD), as two subtypes of coronary artery disease (CAD) in Traditional Chinese Medicine (TCM), following percutaneous coronary intervention (PCI).

METHODS: In this study, a total of 227 CAD patients with QS and 211 CAD patients with QD were enrolled; all participants underwent PCI. Label-free quantification proteomics were employed to analyze the changes in serum in two subtypes of CAD patients before and 6 months after PCI, aiming to elucidate the intervention mechanism of PCI in treating CAD characterized by two different TCM syndromes.

RESULTS: Biochemical analysis revealed significant changes in tumor necrosis factor-α, high density lipoprotein cholesterol, blood stasis clinical symptoms observation, and Gensini levels in both patient groups post-PCI; Proteomic analysis identified 79 and 95 differentially expressed proteins in the QS and QD patient groups, respectively, compared to their control groups. complement C8 alpha chain, complement factor H, apolipoprotein H, apolipoprotein B, plasminogen, carbonic anhydrase 2, and complement factor I were altered in both comparison groups. Furthermore, enrichment analysis demonstrated that cell adhesion and connectivity-related processes underwent changes in QS patients post-PCI, whereas lipid metabolism-related pathways, including the peroxisome proliferator-activated receptor signaling pathway and extracellular matrix receptor interaction, underwent changes in the QD group. The protein-protein interaction network analysis further enriched 52 node proteins, including apolipoprotein B, lipoprotein (a), complement C5, apolipoprotein A4, complement C8 alpha chain, complement C8 beta chain, complement C8 gamma chain, apolipoprotein H, apolipoprotein A-Ⅱ, albumin, complement C4-B, apolipoprotein C3, among others. The functional network of these proteins is posited to contribute to the pathophysiology of CAD characterized by TCM syndromes.

CONCLUSION: The current quantitative proteomic study has preliminarily identified biomarkers of CAD in different TCM subtypes treated with PCI, potentially laying the groundwork for understanding the protein profiles associated with the treatment of various TCM subtypes of CAD.

Key words: percutaneous coronary intervention, proteomics, peptide mapping, coronary atherosclerotic heart disease, Traditional Chinese Medicine syndrome

WANG Zhibo, LI Ying, WANG Daoping, MA Bo, MIAO Lan, REN Junguo, LIU Jinghua, LIU Jianxun. Proteomics analysis of coronary atherosclerotic heart disease with different Traditional Chinese Medicine syndrome types before and after percutaneous coronary intervention[J]. Journal of Traditional Chinese Medicine, 2024, 44(3): 554-563.

Figures/Tables 5

Table 1 Baseline characteristics of the study participants

Item	QD (n = 211)	QS (n = 227)
Age (years)^a	60.3±9.0	60.8±9.1
Males [n (%)]	149 (70.6)	183 (80.6)
Body mass index (kg/m²)^a	24.9±3.2	25.0±2.9
Systolic pressure (mm Hg)^a	130.2±20.4	131.0±20.9
Diastolic pressure (mm Hg)^a	74.9±12.3	75.8±12.8
Resting heart rate (bpm)^a	72.0±11.0	72.0±13.0
Unstable angina [n (%)]	123 (58.3)	151 (66.5)
Non-ST-segment elevation myocardial infarction [n (%)]	32 (15.2)	24 (10.6)
ST-segment elevation myocardial infarction [n (%)]	56 (26.5)	52 (22.9)
History of hypertension [n (%)]	127 (60.2)	136 (59.9)
History of hyperlipidemia [n (%)]	59 (28.0)	81 (35.7)
History of diabetes [n (%)]	67 (31.8)	74 (32.6)
History of stroke [n (%)]	15 (7.1)	23 (10.1)
History of smoking [n (%)]	99 (46.9)	116 (51.1)
Ejection fraction [n (%)]	62.2±9.2	62.5±7.4
Left ventricular internal dimension in diastole (mm)^a	47.7±5.2	47.8±4.8
Erythrocyte (×10¹²/L)^b	4.6 (4.3-4.9)	4.6 (4.3-5.0)
Hemoglobin (g/L)^a	139.3±16.0	141.1±15.7
White blood cell (×10⁹/L)^a	7.6±2.7	7.4±2.7
Percentages of mature neutrophils (%)^b	63.7 (55.6-71.7)	63.1 (57.0-70.8)
Platelet count (×10⁹/L)^a	216.4±55.7	210.9±60.7
Uric acid (μmol/L)^a	347.8±98.2	355.2±96.3
Total cholesterol (mmol/L)^a	4.3±1.1	4.1±1.1
Triglycerides (mmol/L)^b	1.4 (1.0-2.1)	1.39 (1.0-2.0)
Prothrombin time (s)^b	11.3 (10.6-12.5)	11.3 (10.6-12.5)
Activated partial thromboplastin time (s)^a	34.7±15.9	34.1±18.2
D-dimer (ng/mL)^a	101.6±54.3	102.4±36.6
Fibrinogen (μg/mL)^b	3.2 (2.8-3.7)	3.1 (2.6-3.7)

Table 2 Disease indicators in patients with two syndrome types ($\bar{x} \pm s$)

Variable	QSA	QSB	P value	QDA	QDB	P value
TNF-α	4.98±3.08^a	6.56±3.07	<0.01	5.14±2.91^c	7.32±4.34	<0.01
AST	25.25±11.54	28.67±15.01	0.06	23.59±7.37^c	28.56±13.74	<0.01
HDL-C	1.21±0.24^a	1.04±0.26	<0.01	1.17±0.31^c	1.02±0.27	<0.01
BSCSO	18.03±5.80^a	32.27±12.95	<0.01	17.83±6.32^c	33.53±13.94	<0.01
Gensini	20.12±23.47^a	44.60±31.48	<0.01	21.44±24.17^c	53.51±36.39	<0.01
LDL-C	2.13±0.64^b	2.32±0.75	<0.05	2.25±0.76^c	2.55±1.07	<0.01

Figure 1 Quality control of the data and integrated analysis of DEPs A: Pearson correlation analysis, the replicates in each group are 6, 3 subjects by each sex; B: enrichment and clustering analyses of DEPs, with quantitative data standardized; C: the count of DEPs between two comparison groups; D: volcano plot illustrating the distribution of proteins in two comparison groups: red points indicate upregulated proteins, blue points denote downregulated proteins, and gray points signify proteins not differentially expressed. QSA: CAD patients with Qi stagnation and blood stasis syndrome post-PCI; QSB: CAD patients with Qi stagnation and blood stasis syndrome pre-PCI; QDA: CAD patients with Qi deficiency and blood stasis syndrome post-PCI; QDB: CAD patients with Qi deficiency and blood stasis syndrome pre-PCI. DEPs: differentially expressed proteins; CAD: coronary artery disease; PCI: percutaneous coronary intervention.

Figure 2 GO annotation enrichment analysis A: functional enrichment maps of GO terms for QSA/QSB; B: functional enrichment maps of GO terms for QDA/QDB. Depicted in four concentric circles: outermost circle displays gene item classification with a coordinate ruler, using different colors for various categories. Second circle shows the count of background genes per entry and corresponding P-values; longer bars indicate more genes, and redder hues denote smaller P-values. Third circle presents the count of input list genes found in each category, without differentiation between up- and down-regulated genes. Innermost circle illustrates the rich factor, with each background gridline representing 0.1. QSA: CAD patients with Qi stagnation and blood stasis syndrome post-PCI; QSB: CAD patients with Qi stagnation and blood stasis syndrome pre-PCI; QDA: CAD patients with Qi deficiency and blood stasis syndrome post-PCI; QDB: CAD patients with Qi deficiency and blood stasis syndrome pre-PCI; GO: Gene Ontology; PCI: percutaneous coronary intervention; CAD: coronary artery disease.

Figure 3 Overview of pathway enrichment and PPI analysis A: Bubble charts for KEGG pathway enrichment analysis in QSA/QSB; B: PPI networks for DEPs in QSA/QSB; C: Bubble charts for KEGG pathway enrichment analysis in QDA/QDB; D: PPI networks for DEPs in QDA/QDB. In the bubble chart, the X-axis indicates the enrichment factor, while bubble size and color denote the count of foreground genes and their statistical significance, respectively. The rich factor indicates the ratio of enriched DEPs to all genes annotated in a pathway, with higher values indicating greater enrichment. GPI: glycosylphosphatidylinositol; PPAR: peroxisome proliferator-activated receptor; ECM: extracellular matrix; QSA: CAD patients with Qi stagnation and blood stasis syndrome post-PCI; QSB: CAD patients with Qi stagnation and blood stasis syndrome pre-PCI; QDA: CAD patients with Qi deficiency and blood stasis syndrome post-PCI; QDB: CAD patients with Qi deficiency and blood stasis syndrome pre-PCI. PPI: protein-protein interaction; KEGG: Kyoto Encyclopedia of Genes and Genomes; DEPs: differentially expressed proteins; CAD: coronary artery disease; PCI: percutaneous coronary intervention.

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