Atherosclerosis, a progressive disease characterized by lipid accumulation and inflammatory responses within arterial walls, is a major contributor to cardiovascular diseases (CVDs), including myocardial infarction and stroke (1). The development of atherosclerotic plaques is influenced by dysregulated lipid metabolism, oxidative stress, and immune system activation, leading to endothelial dysfunction and arterial narrowing (2). Conventional lipid profiling, which includes measuring total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglycerides, provides valuable insights into cardiovascular risk but does not fully capture the complexity of lipid-mediated disease mechanisms (3).

Lipidomics, an emerging field of metabolomics, allows for a comprehensive analysis of lipid species and their functional roles in disease pathogenesis (4). By employing high-resolution mass spectrometry and advanced bioinformatics, lipidomic profiling can identify lipid subclasses such as sphingolipids, phospholipids, and glycerolipids, which contribute to atherosclerotic plaque formation and progression (5). Altered lipidomic signatures have been associated with endothelial dysfunction, inflammation, and foam cell formation—key events in atherosclerosis development (6).

Recent studies have highlighted the role of specific lipid species, including ceramides and lysophosphatidylcholines, in promoting vascular inflammation and atherogenesis (7). Furthermore, the phosphatidylcholine (PC) to phosphatidylethanolamine (PE) ratio has been identified as a potential biomarker for cardiovascular risk stratification (8). Despite these advancements, the integration of lipidomics into clinical practice remains limited, necessitating further research to validate lipidomic biomarkers and their prognostic utility in atherosclerosis (9).

This study aims to evaluate the role of lipidomics in understanding the biochemical mechanisms underlying atherosclerosis and identifying potential lipid-based biomarkers for cardiovascular risk assessment. Through comprehensive lipid profiling, this research seeks to provide insights into lipid alterations that contribute to disease progression and offer potential therapeutic targets for cardiovascular prevention and management.

Study Design and Participants

This study was a case-control investigation aimed at assessing lipidomic alterations in individuals with atherosclerosis. A total of 100 patients diagnosed with atherosclerosis and 50 age- and sex-matched healthy controls were recruited from a tertiary care hospital. Participants with metabolic disorders, chronic inflammatory diseases, or on lipid-lowering therapy were excluded to minimize confounding variables. Ethical approval was obtained from the institutional ethics committee, and all participants provided written informed consent before enrollment.

Sample Collection and Preparation

Fasting blood samples (10 mL) were collected from each participant using sterile vacutainers. Serum was separated by centrifugation at 3000 rpm for 10 minutes and stored at -80°C until further analysis. Lipid extraction was performed using the Bligh and Dyer method, ensuring efficient recovery of various lipid classes for subsequent analysis.

Lipidomic Analysis

Lipid profiling was conducted using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). A reverse-phase C18 column was utilized for lipid separation, and the mass spectrometer operated in both positive and negative ionization modes to detect a broad range of lipid species. Internal standards were added to quantify lipid subclasses, including phospholipids, sphingolipids, and neutral lipids.

Data Processing and Statistical Analysis

Raw spectral data were processed using LipidSearch software for lipid identification and quantification. Lipid levels were normalized against total lipid content, and statistical analysis was performed using SPSS version 25.0. Differences between groups were assessed using an independent t-test or Mann-Whitney U test for non-normally distributed data. A p-value of <0.05 was considered statistically significant.

Lipid Profile Alterations in Atherosclerosis

A significant difference was observed in the lipid profiles of atherosclerosis patients compared to healthy controls. Total cholesterol levels were markedly elevated in patients (220 ± 15 mg/dL) compared to controls (180 ± 12 mg/dL, p < 0.001). Similarly, low-density lipoprotein (LDL) levels were significantly higher in the atherosclerotic group (140 ± 10 mg/dL) than in controls (100 ± 8 mg/dL, p < 0.001). In contrast, high-density lipoprotein (HDL) levels were reduced in patients (42 ± 6 mg/dL) compared to controls (55 ± 7 mg/dL, p < 0.05) (Table 1).

Triglyceride and Sphingolipid Alterations

Triglyceride concentrations showed a notable increase in atherosclerotic patients (180 ± 12 mg/dL) compared to controls (130 ± 10 mg/dL, p < 0.001). Additionally, sphingomyelin and ceramide levels were significantly elevated in patients (2.8 ± 0.5 µmol/L and 1.5 ± 0.4 µmol/L, respectively) compared to controls (1.9 ± 0.3 µmol/L and 0.9 ± 0.2 µmol/L, p < 0.01) (Table 1).

Phospholipid Ratio and Cardiovascular Risk

The phosphatidylcholine (PC) to phosphatidylethanolamine (PE) ratio, a potential biomarker of cardiovascular risk, was significantly higher in atherosclerotic patients (1.8 ± 0.3) compared to healthy controls (1.2 ± 0.2, p < 0.01). This suggests a strong correlation between lipidomic alterations and cardiovascular disease progression (Table 1).

These findings indicate that lipidomic profiling can provide valuable insights into the biochemical alterations associated with atherosclerosis, supporting its potential role in early disease detection and risk assessment. ​​

Table 1: Lipid Profile Comparison Between Groups

The findings of this study emphasize the crucial role of lipidomics in understanding the biochemical mechanisms underlying atherosclerosis. Elevated total cholesterol, LDL, and triglyceride levels, along with altered sphingolipid and phospholipid ratios, indicate significant disruptions in lipid metabolism among atherosclerosis patients. These observations align with previous studies demonstrating that dysregulated lipid homeostasis is a key driver of atherosclerotic plaque formation and cardiovascular disease progression (1,2).

Dyslipidemia, particularly elevated LDL levels, has been widely recognized as a major risk factor for atherosclerosis (3). Oxidized LDL triggers endothelial dysfunction, promotes inflammation, and contributes to foam cell formation, which accelerates plaque development (4). In the present study, LDL levels were significantly higher in atherosclerotic patients than in healthy controls, reinforcing the role of LDL in disease pathogenesis. Conversely, HDL, which facilitates cholesterol efflux and exerts anti-inflammatory effects, was found to be lower in the atherosclerotic group, further supporting its protective role against cardiovascular disease (5,6).

Apart from conventional lipid markers, lipidomics provides deeper insights into the role of bioactive lipids such as sphingolipids and phospholipids in atherosclerosis. Elevated sphingomyelin and ceramide levels observed in the present study suggest their involvement in inflammatory pathways and vascular dysfunction. Ceramides, in particular, have been implicated in promoting apoptosis, endothelial activation, and insulin resistance, all of which contribute to atherosclerotic progression (7,8). Previous research has demonstrated that ceramide accumulation correlates with increased cardiovascular risk, making it a potential therapeutic target for lipid-lowering strategies (9).

Phospholipid composition alterations also play a role in atherogenesis. The phosphatidylcholine (PC) to phosphatidylethanolamine (PE) ratio has been identified as a potential marker of metabolic and cardiovascular disorders (10). In this study, a significantly higher PC/PE ratio in atherosclerosis patients suggests an imbalance in lipid membrane composition, which may influence lipid signaling and inflammation. This finding is consistent with reports that PC/PE ratio modifications affect membrane fluidity and contribute to oxidative stress-induced vascular damage (11,12).

Lipidomic analysis is emerging as a valuable tool in precision medicine, allowing for early detection of atherosclerotic changes before clinical manifestations appear. Recent advances in mass spectrometry-based lipidomics have enabled the identification of lipid biomarkers associated with cardiovascular risk, paving the way for targeted therapeutic interventions (13). While conventional lipid profiles remain a cornerstone of cardiovascular risk assessment, incorporating lipidomic biomarkers may enhance disease prediction and management strategies (14).

Despite its promising potential, lipidomics still faces challenges in clinical translation. Standardization of lipidomic methodologies, establishment of reference ranges, and validation of lipid biomarkers in large-scale cohorts are necessary for its widespread application in routine clinical practice (15). Future research should focus on integrating lipidomic data with genomic and proteomic insights to develop comprehensive risk assessment models for atherosclerosis.

This study highlights the significance of lipidomics in understanding the biochemical alterations associated with atherosclerosis. The observed elevations in total cholesterol, LDL, triglycerides, sphingomyelin, and ceramide levels, along with a reduced HDL concentration and an altered PC/PE ratio, suggest critical disruptions in lipid metabolism that contribute to disease progression. These findings reinforce the role of specific lipid subclasses as potential biomarkers for early detection and risk assessment of atherosclerosis.

1. O'Donnell VB, Dennis EA, Wakelam MJO, Subramaniam S. LIPID MAPS: Serving the next generation of lipid researchers with tools, resources, data, and training. Sci Signal. 2019;12(563):eaaw2964.
2. Meikle PJ, Christopher MJ. Lipidomics is providing new insight into the metabolic syndrome and its sequelae. Curr Opin Lipidol. 2011;22(3):210-5.
3. Quehenberger O, Dennis EA. The human plasma lipidome. N Engl J Med. 2011;365(19):1812-23.
4. Stegemann C, Pechlaner R, Willeit P, Langley SR, Mangino M, Mayr U, et al. Lipidomics profiling and risk of cardiovascular disease in the prospective population-based Bruneck study. Circulation. 2014;129(18):1821-31.
5. Laaksonen R, Ekroos K, Sysi-Aho M, Hilvo M, Vihervaara T, Kauhanen D, et al. Plasma ceramides predict cardiovascular death in patients with stable coronary artery disease and acute coronary syndromes beyond LDL-cholesterol. Eur Heart J. 2016;37(25):1967-76.
6. Chaurasia B, Summers SA. Ceramides - lipotoxic inducers of metabolic disorders. Trends Endocrinol Metab. 2015;26(10):538-50.
7. Gencer S, Montecucco F, Imboden H, Vicari M, Beer JH, Lüscher TF, et al. Prognostic value of lipoprotein-associated phospholipase A2 in patients with acute coronary syndromes. Eur Heart J. 2011;32(14):1647-55.
8. Meikle PJ, Summers SA. Sphingolipids and phospholipids in insulin resistance and related metabolic disorders. Nat Rev Endocrinol. 2017;13(2):79-91.
9. Han X. Lipidomics for studying metabolism. Nat Rev Endocrinol. 2016;12(11):668-79.
10. Wang DD, Toledo E, Hruby A, Rosner BA, Willett WC, Sun Q, et al. Plasma Ceramides, Mediterranean Diet, and Incident Cardiovascular Disease in the PREDIMED Trial (Prevención con Dieta Mediterránea). Circulation. 2017;135(21):2028-40.
11. Tarasov K, Ekroos K, Suoniemi M, Kauhanen D, Sylvänne T, Hurme R, et al. Molecular lipids identify cardiovascular risk and are efficiently lowered by simvastatin and PCSK9 deficiency. J Clin Endocrinol Metab. 2014;99(1):E45-52.
12. Hammad SM, Pierce JS, Soodavar F, Smith KJ, Al Gadban MM, Rembiesa B, et al. Blood sphingolipidomics in healthy humans: impact of sample collection methodology. J Lipid Res. 2010;51(10):3074-87.
13. Mielke MM, Bandaru VV, Han D, An Y, Resnick SM, Ferrucci L, et al. Factors affecting longitudinal trajectories of plasma sphingomyelins: the Baltimore Longitudinal Study of Aging. Aging Cell. 2015;14(1):112-21.
14. Bismuth J, Lin P, Yao Q, Chen C. Ceramide: a common pathway for atherosclerosis? Atherosclerosis. 2008;196(2):497-504.
15. Sasset L, Zhang Y, Dunn W, Ozcan S. Lipidomics and transcriptomics analysis of triglyceride accumulation induced by oleic acid in human hepatoma cells. BMC Genomics. 2016;17:127.