Supplementary Material for: Exploring the Potential Role of Palmitoleate as a Risk Factor for Atrial Fibrillation: Findings from a Mendelian Randomization Analysis and a Cross-Sectional Study

Background: Atrial fibrillation (AF) risk factors are not fully understood. Reportedly, a relationship exists between fatty acid intake and AF occurrence. The relationship between fatty acid levels and AF requires further investigation, which could help to elucidate AF risk factors and provide insights into its primary prevention. We investigated the relationship between serum fatty acids and AF using Mendelian randomization (MR) and Gas chromatography-mass spectrometry (GC-MS).
Method: MR was used to investigate the potential effects of fatty acids on AF. Patients hospitalized in the Department of Cardiology between May and June 2024 were enrolled and divided into AF and non-AF groups. Fatty acids were extracted from the serum using the Methanol, n-hexane, methyl tert-butyl ether (MTBE) method and methylated using acetyl chloride. GC-MS analysis was used to separate and quantify these methylated mixtures.
Results: Preliminary MR analysis showed that palmitoleic acid and (2 or 3)-decenoic acid are risk factors for AF, while nonadecanoic, oleic, and α/γ-linolenic acids are protective factors. Palmitoleic acid remained a risk factor for AF (OR=1.35, 95% CI=1.19–1.50) after Bonferroni correction, whereas α/γ-linolenic acid remained a protective factor (OR=0.94, 95% CI=0.89–0.98). GC-MS analysis showed that palmitoleic (p=0.028) and stearic (p=0.005) acid levels were significantly higher in the AF group than in the non-AF group. Receiver-operating characteristics curve analysis for palmitoleic acid showed an area under the curve of 0.727, with a sensitivity and specificity of 100% and 50%, respectively.
Conclusion: Palmitoleic acid concentrations were higher in these patients. Fatty acids, including palmitoleic acid, may influence this disease.