Skip to main content
PMC home page
Nutrients logoLink to Nutrients
. 2018 Oct 20;10(10):1558. doi: 10.3390/nu10101558

Sex-Dimorphic Association of Plasma Fatty Acids with Cardiovascular Fitness in Young and Middle-Aged General Adults: Subsamples from NHANES 2003–2004

Pei-Ling Tsou 1, Chang-Jiun Wu 1,*
PMCID: PMC6213878  PMID: 30347828

Abstract

To explore the potential association of plasma fatty acids (FAs) and cardiovascular fitness level (CVFL), data of 449 subjects from 2003–2004 National Health and Nutrition Examination Survey (NHANES) were analyzed. Among these 249 men and 200 women, aged 20–50 years (33.4 ± 8.4 year, mean ± Standard Deviation), 79 low, 166 moderate and 204 high CVFL were categorized by age- and gender- specific percentile, respectively. Twenty-four fatty acids were quantified from fasting plasma. Higher levels of 2 very long-chain saturated FAs (VLSFAs): Arachidic acid (AR1, C20:0) and Docosanoic acid (DA1, C22:0) as well as 2 n-6 polyunsaturated FAs (PUFAs): Arachidonic acid (AA, C20:4n-6) and Docosatetraenoic acid (DTA, C22:4n-6) were observed in the subjects with low CVFL. Notably this association exists only in men. Estimated maximal oxygen consumption (VO2max), the marker for cardiorespiratory fitness, was used for further regression analysis. After the adjustment of potential confounding factors (age, smoking, hypertension status, body mass index (BMI), insulin resistance status, and C-reactive protein (CRP), AA was the only FA correlated with low VO2max in women; while in men AR1, DA1, AA, and DTA remain negatively associated with VO2max. This preliminary analysis suggests a sex-dimorphic relationship between these plasma VLSFAs and n-6 PUFAs with CVFL and merits further investigation.

Keywords: fatty acids, omega-6 polyunsaturated fatty acid, very long chain saturated fatty acid, cardiovascular disease, cardiovascular fitness, plasma

1. Introduction

Cardiovascular fitness, the ability to carry out moderate to high intensity exercise over a prolonged period of time, is measured as maximal oxygen consumption (VO2max) [1]. Numerous epidemiological studies have indicated an inverse association between cardiovascular fitness and coronary heart disease (CHD) or all-cause mortality in healthy participants [2,3,4,5]. The benefits of higher midlife cardiovascular fitness are reported to extend well into the later part of life, up to 4 decades [6].

Plasma FAs serve as physiologically important energy substrates and their release from the adipose tissue by lipolysis is regulated according to the energy demands of the body. In fact, under physiologic conditions, plasma free FAs provide 60–70% of the adenosine triphosphate required by myocardial metabolism [7]. Moreover, FAs are active components of biological membranes. An elevated level of free fatty acids (FFAs) with subsequent increased β-oxidation in the cardiomyocytes can be detrimental for the heart in numerous ways. For example, increased FFAs can lead to damage to the plasma membrane and disturbances of the ion channels of the cardiomyocytes [7].

While fatty acids are known to play important roles in cardiovascular pathophysiology [8,9,10], limited data are available on the relation between plasma fatty acids and cardiovascular fitness. A large body of literature studying FAs effects on cardiovascular health relies on dietary assessment from food frequency questionnaire instead of direct measurement of FAs from plasma. Dietary assessment of individual fatty acid intake, although been used mostly, do not reflect the inter-individual differences in absorption, de novo synthesis, metabolism and other genetic variations leading to different levels of circulating FAs. Measurement of plasma as an aggregate of both dietary and biological processed of FAs may be more appropriate for assessing FAs effects as biomarkers [11].

Despite increasing appreciation of the values of circulating FAs for their biomedical relevance, many studies measure only the total amount but not individual FA levels. However, since different fatty acids have distinct biochemical properties and can execute various biological and metabolic tasks, it is possible that different FAs have their specific changes under diverse pathophysiological settings. For example, γ-linolenic acid (GLA) and dihomo-γ-linolenic (DGLA) have been shown to be positively associated, whereas levels of linoleic acid (LA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are inversely associated, with biomarkers of inflammation [12,13]. Additionally, levels of palmitoleic acid are reported to be positively associated, while levels of LA are inversely associated, with ischemic stroke and insulin resistance [14,15]. Likewise, while plasma total FAs concentrations are known to be associated with cardiovascular and metabolic diseases, emerging evidence has begun to demonstrate that levels of certain FAs may be more informative regarding cardiovascular situation than total amount of FAs. For instance, low levels of C20:4n-3 (eicosatetraenoic acid) and in particular high levels of C18:1n-7 (vaccenic acid) were significantly associated with total mortality in heart failure population [16]. Specifically, among n-6 PUFAs levels in heart failure patients, there was a decrease in the content of C18:2n-6 (linoleic acid), increased C22:5n-6 (DTA) and C22:4n-6 (DPA n-6) and no changes in C20:4n-6 (AA) [16].

Given the importance of cardio-metabolic functions exerted by plasma FAs, it is possible that some circulating FAs could potentially serve as biomarkers reflecting cardiovascular health status. The primary goal of this study was to examine the potential associations of plasma FAs level with CVFL, a modifiable risk factor that has been linked to long-term morbidity and mortality.

2. Materials and Methods

2.1. Study Population

National Health and Nutrition Examination Survey (NHANES) is a cross-sectional observational study conducted by the National Center for Health Statistics of the Centers for Disease Control and Prevention that uses a stratified, multistage probability design to obtain a nationally representative sample of the U.S. population [17]. In the NHANES 2003–2004 cohort (n = 10,122), 1845 people (age >20 years old) have measurement of plasma concentrations of fatty acids, and 2809 people completed a submaximal treadmill exercise test. Current analyses include men and women who have valid data for both examinations (n = 449).

2.2. Examinations and Laboratory Measurements

2.2.1. Cardiorespiratory Fitness Assessment

Per NHANES protocol [18], a multiple-stage submaximal treadmill exercise test was employed to estimate maximal oxygen consumption (VO2max), the marker for cardiorespiratory fitness. Briefly, the exercise test included a two-minute warm-up on the treadmill, two separate three-minute submaximal exercise stages and a two-minute cool-down. Heart rate associated with known workloads at the end of each stage was used to estimate VO2max. Inclusion criteria for exercise testing were based on medical conditions, medication and physical limitations determined during household interviews, and limited to adults 18–49 years old who did not have any known cardiac conditions. Based on gender and age specific criteria, the estimated VO2max is also categorized and reported as “low”, “moderate” or “high” level of cardiovascular fitness in the present dataset. The reference cut-points used for adults are based on data from the Aerobics Center Longitudinal Study (ACLS) [19]. Low level of coefficients of variance (CV) fitness is defined as an estimated VO2max below the 20th percentile of the ACLS data of the same gender and age group; moderate fitness is defined as a value between the 20th and 59th percentile, and high fitness level is defined as at or above the 60th percentile.

2.2.2. Measurement of Plasma Fatty Acids

Fasting plasma fatty acid concentrations were measured by gas chromatography-mass spectrometry (GC-MS) based on a modification of the method of Lagerstedt et al. [20]. All unsaturated fatty acids are assumed to be in the cis-configuration.

2.2.3. Anthropometric Measures

A wall-mounted stadiometer and a digital floor scale were used to measure height and weight, and calibrated as previously described [21].

2.2.4. Smoking Status

Serum cotinine, a major metabolite of nicotine and a biomarker of tobacco smoke exposure, was measured by an isotope dilution-HPLC (high performance liquid chromatography)/atmospheric pressure chemical ionization tandem mass spectrometry [22]. Individuals with serum cotinine levels >10 ng/mL were coded as being current smokers.

2.2.5. Blood Pressure and Hypertension Status

After resting quietly in a sitting position for 5 min and determining the maximum inflation level, three and sometimes 4 blood pressure (BP) determinations (systolic and diastolic) were taken. The arithmetic mean of 3 or 4 measurements were calculated as mean systolic and diastolic pressure. Hypertension was defined if the mean systolic ≥130 mmHg or the mean diastolic ≥80 mmHg.

2.2.6. Assessment of Lipid Profile, Insulin Resistance Status and C-Reactive Protein

Detailed instructions on specimen collection and processing can be found on the NHANES website [17]. Briefly, blood samples were obtained after a 6- or 9-hour fasting by trained medical personnel in mobile examination centers. Total cholesterol is measured enzymatically in serum or plasma in a series of coupled reactions that hydrolyze cholesteryl esters and oxidize the 3-OH group of cholesterol. HDL (high-density lipoprotein) is measured using direct immunoassay. Triglycerides are measured enzymatically in serum using a series of coupled reactions. LDL (low-density lipoprotein)-cholesterol is calculated from measured values of total cholesterol, triglycerides, and HDL-cholesterol according to the Friedewald calculation:

[LDL−cholesterol]=[total cholesterol]−[HDL−cholesterol]−[triglycerides/5]

where [triglycerides/5] is an estimate of VLDL (very low-density lipoprotein)-cholesterol and all values are expressed in mg/dL. Plasma glucose concentration was determined by hexokinase enzyme method. Plasma insulin concentrations were measured with Tosoh AIA-PACK IRI (Toyama, Japan) two-site immunoenzymometric assay. C-peptide concentrations were measured using 125I-labeled radioimmunoassay. HOMA-IR (homeostatic model assessment for insulin resistance the product of fasting plasma glucose and insulin concentrations divided by 22.5 [mM×μIU/mL/22.5]) was used as a composite index for insulin resistance status. C-reactive Protein (CRP) is quantified by latex-enhanced nephelometry.

2.3. Statistical Methods

Descriptive statistics including means, standard deviations, and range for continuous variables and frequencies for categorical variables were reported. To test if the levels of an individual demographic variable are different among three fitness groups, ANOVA (Analysis of Variance) test was used for continuous variables and Fisher’s exact test was used for categorical variables. A two-tailed p value < 0.05 was considered statistically significant. The differences of plasma FAs between CVFL groups were assessed by ANOVA followed with False Discovery Rate (FDR) estimation for multiple comparisons correction. An FDR value <0.30 was considered statistically significant. Post-hoc pairwise t-test analysis was implemented on significant FAs to understand which groups are different. Univariate and multivariate linear regression models were also implemented with VO2max as dependent variable and each plasma FA as main independent variable. Both VO2max and plasma FA levels were logarithmically transformed due to skewed distribution. In multivariate models, appropriate confounding variables were included. All statistical analyses were conducted using R software (R version 3.3.0, Vienna, Austria).

3. Results

3.1. Subject Characteristics

Table 1 presents the demographic, anthropometric, and biochemical characteristics of the whole study population, as well as information stratified by 3 CVFL groups—“low”, “moderate”, and “high”—with gender and age specific criteria (see Methods 2.2.1). The mean age of the all subjects was 33.4 years and there were slightly more men (55.5%) than women (44.5%). The mean maximal oxygen uptake (VO2max) was 32.5, 38.9, and 48.8 mL/kg/min for men and 26.8, 31.5, and 42.3 mL/kg/min for women categorized as low, moderate, and high CVFL, respectively. The distribution of age and gender among 3 CVFL groups were not significantly different. The percentage of current smoking (defined by serum cotinine level) was similar among 3 CVFL groups. The average waist circumference (WC) and body mass index was 90.6 cm and 25.95 kg/m2 while subjects with lower CVFL tended to have higher WC and BMI. Additionally, both average systolic and diastolic blood pressure were significantly higher in the low CVFL group, though the percentage of hypertension (defined by average SBP ≥130 mmHg or average DBP ≥80 mmHg) only showed a trend without statistical significance. Fasting plasma glucose concentration and lipid profile (including total cholesterol, triglyceride, HDL and LDL) did not differ among 3 CVFL groups. Notably, plasma insulin, C-peptide concentrations and derived insulin resistance status (HOMA-IR) were negatively correlated with CVFL. The level of plasma C-reactive Protein (CRP) did not show difference among 3 groups.

Table 1.

Demographic characteristics of the study population: overall and stratified by cardiovascular fitness levels. CVFLs were categorized into Low, Moderate, and High by estimated VO2max with gender and age specific percentile. Significant values are shown in bold and italic fonts.

All Subjects Cardiovascular Fitness Level (CVFL)
Low Moderate High ANOVA
Range (Mean ± S.D) n Range (Mean ± S.D) n Range (Mean ± S.D) n Range (Mean ± S.D) n p value
VO2max(ml/kg/min) 20.9–78.9 (39.3 ± 8.7) 448 20.9–37.1 (29.8 ± 4) 79 27.1–43.9 (36 ± 4.4) 166 33.1–78.9 (45.7 ± 7.8) 204 <0.001
Age (yr) 20.1–50 (33.4 ± 8.4) 448 20.2–49.9 (31.9 ± 8.3) 78 20.2–49.4 (34.1 ± 7.9) 166 20.1–50 (33.4 ± 8.9) 204 0.161
Gender (M,%) 55.5 53.2 78 60.2 166 52.5 204 ¶0.294
Smoking (%) 29.4 449 25.6 78 32.5 166 28.2 202 0.486
Waist Circumfrence (WC, cm) 66.3–122 (90.6 ± 11.9) 449 68.4–120 (94.0 ± 12.0) 54 66.5–117 (90.5 ± 11.2) 129 66.3–122 (89.5 ± 12.2) 164 0.049
Body Mass Index (kg/m2) 16.7–40.9 (26.0 ± 4.2) 347 17.6–39.5 (27.3 ± 4.1) 54 16.7–40 (25.9 ± 4.2) 130 17.4–40.9 (25.5 ± 4.2) 165 0.026
Hypertension (%) 19.8 349 31.5 54 18.5 130 17.0 165 ¶0.060
Systolic Blood Pressure ( SBP, mmHg) 87–189 (114 ± 11) 349 98–189 (117 ± 14.7) 54 87–143 (114.7 ± 10.8) 130 89–145 (112.7 ± 10.6) 165 0.029
Diastolic Blood Pressure ( DBP, mmHg) 40–95 (69 ± 10) 349 51–93 (72.6 ± 10.5) 54 40–95(69.52 ± 9.49) 130 44–95 (68.6 ± 10.1) 165 0.037
Plasma Triglyceride (TG, mg/dL ) 32–1779 (131 ± 132) 347 32–776 (136 ± 105) 54 35–648 (134. ± 96) 129 33–1779 (128 ± 161) 164 0.892
Plasma HDL (mg/dL) 27–112 (54.6 ± 15.9) 348 29–102 (54.9 ± 17.6) 54 27–103 (52.2 ± 13.4) 130 27–112 (56.5 ± 17) 164 0.065
Plasma LDL (mg/dL) 35–217 (115 ± 33.6) 435 35–201 (118 ± 32.6) 78 41–211 (115 ± 34.6) 160 41–217 (113 ± 33.1) 197 0.466
Plasma Total Cholesterol (mg/dL) 93–426 (193 ± 39.4) 447 93–426 (199 ± 44.1) 79 109–295 (192 ± 39.2) 166 99–306 (192 ± 37.6) 202 0.390
Plasma Glucose (GLU, mg /dL) 63.4–416 (93.5 ± 19.8) 449 72.6–416 (98.3 ± 44.9) 54 75.6–142 (93.7 ± 8.9) 130 63.4–165 (91.9 ± 10.6) 165 0.115
Plasma C-peptide ( nmol/L ) 0.71–71.1 (9.8 ± 8.8) 446 2.17–42.5 (12.0 ± 8.5) 79 1.16–71.1 (9.9 ± 9) 166 0.71–54.5 (8.9 ± 8.5) 201 0.023
Plasma Insulin( INS, μU/mL ) 0.02–2.77 (0.73 ± 0.35) 446 0.3–2.1 (0.83 ± 0.36) 79 0.24–2.77 (0.74 ± 0.35) 166 0.02–2.47 (0.68 ± 0.34) 201 0.005
HOMA-IR 0.12–20 (2.36 ± 2.33) 446 0.47–11.3 (2.90 ± 2.24) 79 0.25–20 (2.39 ± 2.31) 166 0.12–16.7 (2.13 ± 2.36) 201 0.044
Plasma C-Reactive Protein (CRP, mg/dL) 0.01–16.5 (0.36 ± 0.9) 448 0.02–2.91 (0.44 ± 0.53) 79 0.01–3.41 (0.33 ± 0.42) 166 0.01–16.50 (0.36 ± 1.24) 203 0.670
Open in a new tab

Abbreviations: VO2max: Estimated maximal oxygen uptake; HOMA-IR: Homeostatic Model Assessment - Insulin Resistance; ANOVA: Analysis of Variance; n: number. ¶ Fisher’s exact test.

3.2. Twenty-four Fatty Acid Levels in Fasting Plasma

Plasma concentrations of 24 FAs, including 6 SFAs, 7 MUFAs, and 11 PUFAs from all 449 subjects and levels of 3 CVFL groups and are listed in Table 2. All 24 FAs, including the so-called heart healthy n-3 FAs, exhibited negative relationships with CVFL, although only 2 SFAs and 3 n-6 PUFAs reached statistical significance. Two VLSFA, Arachidic acid (AR1, C20:0) and Docosanoic acid (DA1, C22:0) as well as one n-6 PUFA, Arachidonic acid (AA, C20:4n-6), were inversely associated with CVFL (ANOVA FDR < 0.30). None of the derivative variables of interest, including the sum of 24 FAs, the sum of 6 SFAs, the sum of 7 MUFA, the sum of 11 PUFA, the sum of 5 n-3 PUFAs, the sum of 6 n-6 PUFA, the ratio of n-6 to n-3 PUFA, or the ratio of C16:1 to C16:0 (Δ9 desaturase index, a surrogate marker of the activity of Stearoyl-coenzyme A desaturase) showed significance difference among the 3 groups.

Table 2.

Twenty-four Fatty Acid Levels in Fasting Plasma and 8 derivative variables of interest: overall and stratified by cardiovascular fitness levels

All Subjects Cardiovascular Fitness Level (CVFL)
Low Moderate High ANOVA
Range (Mean ± S.D) n Range (Mean ± S.D) n Range (Mean ± S.D) n Range (Mean ± S.D) n p value FDR
SFA C14:0 Myristic Acid 24.8–2410 (137 ± 139) 442 40.2–370 (130 ± 72) 79 24.8–620 (138 ± 93.6) 164 29.2–2410 (140 ± 185) 199 0.88 1.00
C16:0 Palmitic Acid 1310–8240 (2715 ± 960) 444 1310–8240 (2840 ± 990) 79 1380–7120 (2767 ± 996) 165 1330–7970 (2623 ± 914) 200 0.16 1.00
C18:0 Stearic Acid 364–1720 (685 ± 180) 444 404–1720 (708 ± 186) 79 364–1470 (694 ± 182) 165 373–1490 (669 ± 175) 200 0.19 1.00
C20:0 Arachidic Acid (AR1) 6.25–83.7 (23 ± 5.8) 433 16.1–83.7 (24.9 ± 8.1) 78 12.4–36.3 (22.6 ± 5) 159 6.25–43.7 (22.6 ± 5.1) 196 0.007 0.23
C22:0 Docosanoic Acid (DA1) 17–171 (68.3 ± 16.5) 431 38.2–171 (73.5 ± 18.2) 78 38–129 (66.9 ± 16.4) 158 17–117 (67.5 ± 15.6) 195 0.009 0.26
C24:0 Lignoceric Acid 20.9–132 (54.2 ± 13.3) 430 27.1–112 (56.5 ± 13.7) 76 28.8–132 (53.1 ± 13.2) 161 20.9–104 (54.1 ± 13.2) 193 0.18 1.00
MUFA C14:1 n-5 Myristoleic Acid 0.70–186 (8.8 ± 11.9) 445 0.98–29.4 (7.8 ± 5.3) 79 0.9–75.7 (9.2 ± 10) 165 0.70–186 (8.9 ± 14.8) 201 0.69 1.00
C16:1 n-7 Palmitoleic Acid 39–2500 (238 ± 199) 445 57.7–645 (232 ± 122) 79 51.9–1930 (245 ± 198) 165 39–2500 (236 ± 224) 201 0.86 1.00
C18:1 n-7 cis-Vaccenic Acid 54.1–550 (144 ± 59.1) 435 62.9–550 (148 ± 60.4) 78 59.7–489 (146 ± 57.8) 161 54.1–454 (141 ± 59.7) 196 0.6 1.00
C18:1 n-9 Oleic Acid 885–15800 (2107 ± 1175) 443 885–8560 (2111 ± 926) 79 933–13300 (2157 ± 1193) 164 911–15800 (2065 ± 1249) 200 0.76 1.00
C20:1 n-9 Eicosenoic Acid 2.89–61.1 (13.8 ± 6.7) 443 4.76–61.1 (14.2 ± 7.3) 79 5.09–42 (14 ± 6.8) 164 2.89–46.3 (13.6 ± 6.3) 200 0.72 1.00
C22:1 n-9 Docosenoic Acid 0.21–23.8 (4.5 ± 3.5) 395 0.21–23.8 (4.9 ± 3.7) 65 0.21–18.2 (4.3 ± 3.3) 145 0.21–23.80 (4.39 ± 3.60) 185 0.48 1.00
C24:1 n-9 Nervonic Acid 18.3–162 (76 ± 18.8) 428 38.6–133 (79.3 ± 19.4) 76 34.1–162 (76.7 ± 20.1) 157 18.3–117 (74.1 ± 17.4) 195 0.1 1.00
PUFA C18:2 n-6 Linoleic Acid (LA) 1770–9200 (3503 ± 881) 445 1810–9200 (3571 ± 1000) 79 2130–6970 (3454 ± 844) 165 1770–7450 (3517 ± 863) 201 0.6 1.00
C18:3 n-6 gamma-Linolenic Acid (GLA) 6.25–141 (48.4 ± 23) 442 7.2–136 (48.8 ± 26) 79 10.50–141 (49.2 ± 23.2) 164 6.25–122 (47.5 ± 21.7) 199 0.77 1.00
C20:2 n-6 Eicosadienoic Acid 5.66–123 (22.4 ± 10.3) 443 9.9–52 (22.6 ± 8.5) 79 8.3–64.9 (22.2 ± 9.1) 164 5.66–123 (22.5 ± 11.7) 200 0.92 1.00
C20:3 n-6 Dihomo-gamma-Linolenic Acid (DGLA) 49–716 (156 ± 63.9) 445 53.1–330 (158 ± 51.3) 79 49–612 (158.34 ± 64.57) 165 50.9–716 (153 ± 67.8) 201 0.71 1.00
C20:4 n-6 Arachidonic Acid (AA) 341–1770 (784 ± 205) 445 443–1770 (839 ± 232) 79 380–1390 (796 ± 201) 165 341–1590 (754 ± 194) 201 0.005 0.16
C22:4 n-6 Docosatetraenoic Acid (DTA) 6.72–66.1 (26.2 ± 8.7) 445 13.3–63.3 (28.2 ± 9.5) 79 11.3–66.1 (26.8 ± 9) 165 6.72–58.8 (25 ± 8.1) 201 0.013 0.37
C22:5 n-6 Docosapentaenoic-6 Acid (DPA n-6) 5.31–61.7 (21.5 ± 8.9) 445 9.5–52.5 (24 ± 9.4) 79 5.94–56 (21.5 ± 9.1) 165 5.31–61.7 (20.5 ± 8.3) 201 0.013 0.37
C18:3 n-3 alpha-Linolenic Acid (ALA) 16.1–421 (68.2 ± 42.1) 445 23.1–341 (67.1 ± 44) 79 16.1–272 (67.9 ± 37.4) 165 20.2–421 (68.8 ± 45) 201 0.95 1.00
C20:5 n-3 Eicosapentaenoic Acid (EPA) 6.–236 (43.4 ± 28.8) 444 15.7–130 (41.3 ± 23.9) 79 6.60–236 (45.5 ± 33.8) 165 6.03–143 (42.4 ± 26.1) 200 0.46 1.00
C22:5 n-3 Docosapentaenoic-3 Acid (DPA n-3) 12.1–106 (41.5 ± 13.6) 445 17.4–106 (41.4 ± 14.8) 79 15.1–90.4 (42.5 ± 14.2) 165 12.1–98.5 (40.6 ± 12.6) 201 0.42 1.00
C22:6 n-3 Docosahexaenoic Acid (DHA) 43.2–549 (131 ± 62.6) 445 55.1–468 (134 ± 61.3) 79 46.6–549 (135 ± 70.8) 165 43.2–324 (126 ± 55.5) 201 0.35 1.00
Σ24FFA 13187–78613 (27060 ± 7739) 445 13981–78613 (27800 ± 8222) 79 15832–67736 (27218 ± 7729) 165 13187–67035 (26639 ± 7563) 201 0.5 1.00
Σ6SFA 1840–10670 (3669 ± 1216) 445 1851–10677 (3829 ± 1246) 79 1977–9243 (3735 ± 1260) 165 1840–10323 (3552 ± 1162) 201 0.16 1.00
Σ7MUFA 345–17590 (2577 ± 1388) 445 1129–9823 (2592 ± 1078) 79 345–14755 (2631 ± 1421) 165 480–17590 (2527 ± 1470) 201 0.77 1.00
Σ11PUFA 2375–12529 (4845 ± 1124) 445 2662–12529 (4975 ± 1288) 79 2891–8697 (4818 ± 1075) 165 2375–9739 (4816 ± 1096) 201 0.52 1.00
Σw3PUFA ¶ 92.6–1071 (284 ± 119) 445 142–1030 (284 ± 118) 79 92.6–1071 (291 ± 129) 165 107–786 (278 ± 110) 201 0.57 1.00
Σw6PUFA § 2255–11499 (4561.6 ± 1048) 445 2472–11499 (4692 ± 1199) 79 2781–8229 (4528 ± 1002) 165 2255–8953 (4539 ± 1024) 201 0.47 1.00
w6/w3 5.29–37.7 (17.5 ± 4.7) 445 8.2–31.6 (17.7 ± 4.6) 79 5.29–34.4 (17.1 ± 4.7) 165 8.02–37.7 (17.8 ± 4.9) 201 0.41 1.00
C16:1/C16:0 1.82–37.4 (14.1 ± 5.1) 444 5.99–24.1 (13.8 ± 4.2) 79 3.69–33.8 (13.8 ± 4.9) 165 1.82–37.4 (14.3 ± 5.5) 200 0.62 1.00
Open in a new tab

All values are in μmol/L. Significant values are shown in bold and italic fonts. Abbreviations: SFA: Saturated Fatty Acid; MUFA: Mono-Unsaturated Fatty Acid; PUFA: Poly-Unsaturated Fatty Acid. ¶ Σw3PUFA: Σ (SSALN_N, SSEPA_N, SSDP3_N, SSDHA_N ). Σ6SFA: Sum of 6 SFAs; Σ7MUFA: Sum of 7 MUFAs; Σ11PUFA: Sum of 11 PUFAs; Σ24FFA: Σ6SFA + Σ7MUFA + Σ11PUFA. ¶ Σw3PUFA: Σ (SSALN_N, SSEPA_N, SSDP3_N, SSDHA_N ); § Σw6PUFA: Σ (SSLNA_N, SSGLA_N, SSED1_N, SSHGL_N, SSARA_N, SSDTA_N, SSDP6_N).

3.3. FAs Levels among 3 CV Fitness Groups Stratified by Sex

Next, we set out to determine whether the relationships of FAs and CVFLs were different in men and women. The levels of 24 FAs among 3 CV fitness groups were further stratified by sex and listed in Table 3. Notably, none of the 24 FAs showed significant difference among 3 CVFL groups in women. On the other hand, the negative associations of CVFL with 2 VLSFAs (AR1, DA1) and one n-6 PUFA (AA) were even stronger in the analysis of men then in the whole 449 subjects (Table 3 and Figure 1). Additionally, the negative association of CVFL with Docosatetraenoic acid (DTA, C22:4n-6) could only be revealed when examined the levels specifically in men but not from the entire population.

Table 3.

Twenty-four Fatty Acid Levels in Fasting Plasma and 8 derivative variables of interest among 3 cardiovascular levels further stratified by sex.

Men Women
Cardiovascular Fitness Level Cardiovascular Fitness Level
Low Moderate High ANOVA Low Moderate High ANOVA
mean ± S.D n mean ± S.D n mean ± S.D n p FDR mean ± S.D n mean ± S.D n mean ± S.D n p FDR
SFA C14:0 Myristic Acid 141 ± 78.5 42 141 ± 103 99 134 ± 105 103 0.88 1.00 118 ± 62 37 133 ± 77.9 65 145 ± 244 96 0.71 1.00
C16:0 Palmitic Acid 2999 ± 1098 42 2769 ± 1114 100 2678 ± 971 105 0.25 1.00 2659 ± 828 37 2764 ± 789 65 2561 ± 847 95 0.31 1.00
C18:0 Stearic Acid 756 ± 212 42 694 ± 203 100 685 ± 185 105 0.14 1.00 654 ± 134 37 694 ± 147 65 640 ± 161 95 0.18 1.00
C20:0 Arachidic Acid (AR1) 26.3 ± 10.4 41 21.8 ± 4.8 99 21.8 ± 4.7 102 <0.001 0.004 23.2 ± 3.8 37 23.9 ± 5.0 60 23.5 ± 5.4 94 0.77 1.00
C22:0 Docosanoic Acid (DA1) 76.7 ± 21.1 41 64.4 ± 16.2 99 65.8 ± 15.6 101 <0.001 0.01 70.0 ± 13.8 37 71.0 ± 15.9 59 69.2 ± 15.4 94 0.78 1.00
C24:0 Lignoceric Acid 60.8 ± 14.7 40 53.7 ± 14.2 100 55.9 ± 13.4 102 0.028 0.75 51.7 ± 10.8 36 52.0 ± 11.3 61 52.2 ± 12.8 91 0.98 1.00
MUFA C14:1 n-5 Myristoleic Acid 8.3 ± 5.5 42 9.9 ± 12.0 100 8.4 ± 9.3 105 0.48 1.00 7.3 ± 5.1 37 8.1 ± 5.8 65 9.4 ± 19.1 96 0.69 1.00
C16:1 n-7 Palmitoleic Acid 244 ± 125 42 255 ± 237 100 246 ± 271 105 0.95 1.00 219 ± 118 37 229 ± 112 65 226 ± 169 96 0.93 1.00
C18:1 n-7 cis-Vaccenic Acid 160 ± 75.5 41 147 ± 64.1 99 143 ± 62.4 103 0.37 1.00 135 ± 33.4 37 145 ± 46.4 62 139 ± 56.8 93 0.58 1.00
C18:1 n-9 Oleic Acid 2290 ± 1147 42 2241 ± 1441 100 2095 ± 846 104 0.55 1.00 1908 ± 529 37 2026 ± 631 64 2032 ± 1577 96 0.85 1.00
C20:1 n-9 Eicosenoic Acid 14.9 ± 8.7 42 14.0 ± 7.3 100 13.6 ± 6.4 105 0.62 1.00 13.4 ± 5.4 37 14.1 ± 5.9 64 13.5 ± 6.2 95 0.81 1.00
C22:1 n-9 Docosenoic Acid 5.3 ± 4.4 35 4.2 ± 3.1 88 4.5 ± 4.1 100 0.33 1.00 4.5 ± 2.4 30 4.5 ± 3.5 57 4.2 ± 3.0 85 0.78 1.00
C24:1 n-9 Nervonic Acid 81.1 ± 22.4 39 75.5 ± 20.6 98 72.9 ± 17.8 103 0.086 1.00 77.5 ± 15.6 37 78.5 ± 19.3 59 75.6 ± 16.9 92 0.58 1.00
PUFA C18:2 n-6 Linoleic Acid (LA) 3725 ± 1127 42 3308 ± 806 100 3516 ± 845 105 0.029 0.76 3397 ± 814 37 3679 ± 859 65 3518 ± 887 96 0.26 1.00
C18:3 n-6 gamma-Linolenic Acid (GLA) 57.9 ± 29.6 42 52.6 ± 25.1 99 53.3 ± 21.4 103 0.47 1.00 38.3 ± 15.9 37 44.1 ± 19.1 65 41.4 ± 20.4 96 0.33 1.00
C20:2 n-6 Eicosadienoic Acid 22.5 ± 8.0 42 20.8 ± 7.9 100 21.4 ± 9.1 105 0.56 1.00 22.7 ± 9.2 37 24.2 ± 10.5 64 23.8 ± 14.0 95 0.84 1.00
C20:3 n-6 Dihomo-gamma-Linolenic Acid (DGLA) 158 ± 50 42 153 ± 68.4 100 153 ± 57.4 105 0.92 1.00 159 ± 53.4 37 166 ± 57.7 65 153 ± 77.9 96 0.48 1.00
C20:4 n-6 Arachidonic Acid (AA) 891 ± 255 42 790 ± 190 100 770 ± 203 105 0.006 0.17 780 ± 188 37 805 ± 219 65 735 ± 182 96 0.079 1.00
C22:4 n-6 Docosatetraenoic Acid (DTA) 31.6 ± 10 42 27.3 ± 9.1 100 26.2 ± 8.5 105 0.005 0.15 24.5 ± 7.3 37 25.9 ± 8.7 65 23.8 ± 7.4 96 0.23 1.00
C22:5 n-6 Docosapentaenoic-6 Acid (DPA n-6) 23.9 ± 9 42 20.6 ± 8.6 100 19.9 ± 8 105 0.037 0.89 24.1 ± 9.9 37 22.9 ± 9.6 65 21.1 ± 8.6 96 0.2 1.00
C18:3 n-3 alpha-Linolenic Acid (ALA) 73.1 ± 53.2 42 66.8 ± 39.0 100 70.0 ± 49.1 105 0.75 1.00 60.3 ± 29.6 37 69.6 ± 34.9 65 67.4 ± 40.3 96 0.46 1.00
C20:5 n-3 Eicosapentaenoic Acid (EPA) 46.1 ± 23.3 42 47.8 ± 34.7 100 45.5 ± 27.5 104 0.85 1.00 35.8 ± 23.7 37 42.0 ± 32.3 65 39.1 ± 24.2 96 0.54 1.00
C22:5 n-3 Docosapentaenoic-3 Acid (DPA n-3) 47.5 ± 16.1 42 43.9 ± 15.0 100 43.1 ± 12.9 105 0.25 1.00 34.5 ± 9.5 37 40.4 ± 12.6 65 38.0 ± 11.8 96 0.054 1.00
C22:6 n-3 Docosahexaenoic Acid (DHA) 130 ± 67.6 42 125 ± 59.8 100 120 ± 52.8 105 0.66 1.00 139 ± 53.9 37 151 ± 83.1 65 132 ± 57.9 96 0.23 1.00
Σ24FFA 29343 ± 9546 42 26962 ± 8491 100 26932 ± 7471 105 0.23 1.00 26050 ± 6061 37 27611 ± 6426 65 26318 ± 7688 96 0.43 1.00
Σ6SFA 4054 ± 1395 42 3741 ± 1409 100 3634 ± 12412 105 0.23 1.00 35734 ± 1010 37 3725.7 ± 996.2 65 3463 ± 1067 96 0.29 1.00
Σ7MUFA 2793 ± 1322 42 2743 ± 1697 100 25597 ± 1125 105 0.54 1.00 2364 ± 655 37 2460 ± 820 65 2492 ± 1779 96 0.89 1.00
Σ11PUFA 5206 ± 1461 42 4654 ± 1041 100 4838 ± 1091 105 0.033 0.83 4715 ± 1015 37 5070 ± 1087 65 4793 ± 1106 96 0.18 1.00
Σw3PUFA ¶ 296 ± 138 42 283 ± 122 100 279 ± 115 105 0.72 1.00 269 ± 91.2 37 303 ± 139 65 277 ± 106 96 0.27 1.00
Σw6PUFA § 4910 ± 1347 42 4371 ± 968 100 4560 ± 1014 105 0.023 0.64 4445 ± 965 37 4767 ± 1012 65 4516 ± 1040 96 0.2 1.00
w6/w3¶ 17.8 ± 4.9 42 17.1 ± 4.8 100 17.9 ± 5.0 105 0.5 1.00 17.5 ± 4.4 37 17.1 ± 4.5 65 17.6 ± 4.8 96 0.79 1.00
C16:1/C16:0 14.1 ± 4.6 42 14.0 ± 5.3 100 14.6 ± 6.0 105 0.72 1.00 13.6 ± 3.7 37 13.6 ± 4.2 65 14.0 ± 4.9 95 0.81 1.00
Open in a new tab

All values are in μmol/L. Significant values are shown in bold and italic fonts. Abbreviations: SFA: Saturated Fatty Acid; MUFA: Mono-Unsaturated Fatty Acid; PUFA: Poly-Unsaturated Fatty Acid. ¶ Σw3PUFA: Σ (SSALN_N, SSEPA_N, SSDP3_N, SSDHA_N ). ¶ Σw3PUFA: Σ (SSALN_N, SSEPA_N, SSDP3_N, SSDHA_N ) § Σw6PUFA: Σ (SSLNA_N, SSGLA_N, SSED1_N, SSHGL_N, SSARA_N, SSDTA_N, SSDP6_N).

Figure 1.

Figure 1

Open in a new tab

Boxplots of concentrations of 4 FAs (A: Arachidic acid, B: Docosanoic acid, C: Arachidonic Acid, and D: Docosatetraenoic Acid) with significant association with CVFL stratified by sex. CVFLs were categorized into low (white), moderate (light gray), and high (dark gray). The levels of Men and Women were shown at the left and right side, respectively. In addition to ANOVA test among 3 CVFL groups, post-hoc pairwise t-test analysis was implemented in Men. Significant t-test p values comparing Low vs. Moderate + High CVFLs in men were shown in the figures. FAs: fatty acids; CVFL: cardiovascular fitness level; ANOVA: Analysis of Variance; FDR: false discovery rate.

3.4. Subjects with Low CVFL Have Higher AR1, DA1, AA, and DTA

Notably, as shown in Figure 1 and Table 4, the levels of Arachidic acid (AR1, C20:0) and Docosanoic acid (DA1, C22:0) were higher only in the low CVFL group, whereas the levels were similar in the moderate and high CVFL groups. As seen in Table 3, for AR1, the mean ± S.D was 26.3 ± 10.4 in the low group, 21.8 ± 4.8 and 21.8 ± 4.7 for the moderate and high CVFLs groups, respectively. For DA1, the mean ± S.D was 76.7 ± 21.1 in the low group, 64.4 ± 16.2 and 65.8 ± 15.6 for the moderate and high CVFLs groups, respectively.

Table 4.

ANOVA and pairwise t-test p values comparing Low vs. Moderate + High CVFLs in men.

ANOVA p t-test p
L vs. M vs. H L vs. M L vs. H M vs. H L vs. (M + H)
Arachidic acid <0.001 0.01 0.01 0.998 0.01
Docosanoic acid <0.001 0.001 0.004 0.545 0.002
Arachidonic Acid 0.006 0.024 0.008 0.481 0.01
Docosatetraenoic Acid 0.005 0.021 0.003 0.346 0.037
Open in a new tab

Abbreviations: ANOVA (Analysis of Variance); L: low; M: moderate; H: high.

On the other hand, for n-6 PUFAs (AA and DTA), there seemed a trend of negative associations across the 3 groups. For AA, the mean ± S.D was 891 ± 255 in the low group, 790 ± 190 in the moderate and 770 ± 203 in the high CVFLs groups. For DTA, the mean ± S.D was 31.6 ± 10 in the low group, 27.3 ± 9.1 in the moderate and 26.2 ± 8.5 in the high CVFLs groups. However, the difference between the moderate and the high CVFL group did not reach statistic significance (p = 0.48 for AA, and p = 0.35 for DTA, respectively). Thus, higher levels of these 4 FAs were seen only in the low CVFL group.

3.5. Univariate and Multivariate Linear Regression Analysis

To further study the relationships of FAs and CVFL, we used VO2max as outcome variable and individual FA levels as independent variable in linear regression models. The 3 CVFLs based on estimated VO2max is gender and age adjusted whereas the estimated VO2max itself is not adjusted. Potential confounding variables were selected taking into account univariate analysis of known factors that reported to be associated CVFLs (data not shown). The effects of various combinations of potential confounding factors were adjusted with multivariate models as described in Table 5. For men, consistently, the negative association of Arachidic acid (AR1), Docosanoic acid (DA1), Arachidonic acid (AA), and Docosatetraenoic acid (DTA) remained significant in all models. For women, on the other hand, Arachidonic acid (AA) was the only FA that showed a negative association with VO2max in multivariate model after adjusting confounding factors.

Table 5.

Univariate and multivariate linear regression analysis in Men and Women separately.

Men
SSAR1_N C20:0 Arachidic Acid
SSAR1_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.235 6e-7 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.161 0.002 −0.004 0.010 0.002 0.94 −0.003 0.025 −0.001 0.64 --- --- --- --- --- ---
model2 −0.161 0.002 −0.004 0.011 0.002 0.95 −0.003 0.025 −0.001 0.64 −0.001 0.94 --- --- --- ---
model3 −0.154 0.003 −0.004 0.007 0.002 0.94 −0.003 0.038 −0.0004 0.92 −0.001 0.91 −0.007 0.36 --- ---
model4 −0.167 0.004 −0.004 0.009 0.0004 0.99 −0.003 0.036 −0.001 0.84 −0.001 0.92 −0.008 0.33 6.6e-5 0.60
SSDA1_N C22:0 Docosanoic Acid
SSDA1_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.182 6e-5 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.125 0.012 −0.004 0.008 −0.002 0.94 −0.003 0.016 −0.002 0.48 --- --- --- --- --- ---
model2 −0.126 0.012 −0.004 0.008 −0.002 0.93 −0.003 0.017 −0.002 0.49 −0.003 0.77 --- --- --- ---
model3 −0.123 0.014 −0.004 0.004 −0.002 0.95 −0.003 0.029 −0.001 0.84 −0.003 0.74 −0.009 0.23 --- ---
model4 −0.121 0.017 −0.004 0.007 −0.001 0.98 −0.003 0.030 −0.0004 0.91 −0.003 0.74 −0.008 0.33 −5.5e-5 0.64
SSARA_N C20:4 n-6 Arachidonic Acid (AA)
SSARA_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.178 4e-5 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.119 0.018 −0.004 0.008 0.012 0.65 −0.003 0.039 −0.002 0.53 --- --- --- --- --- ---
model2 −0.119 0.018 −0.004 0.008 0.012 0.65 −0.003 0.039 −0.002 0.53 0.0002 0.98 --- --- --- ---
model3 −0.115 0.022 −0.004 0.005 0.012 0.64 −0.002 0.064 −0.0004 0.90 −0.0002 0.98 −0.01 0.21 --- ---
model4 −0.117 0.026 −0.004 0.008 0.012 0.65 −0.002 0.062 −0.0004 0.91 −0.0002 0.98 −0.009 0.28 −1.9e-5 0.88
SSDTA_N C22:4 n-6 Docosatetraenoic Acid (DTA)
SSDTA_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.123 3e-4 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.084 0.021 −0.004 0.004 0.011 0.66 −0.003 0.027 −0.002 0.50 --- --- --- --- --- ---
model2 −0.084 0.021 −0.004 0.004 0.011 0.67 −0.003 0.027 −0.002 0.50 −0.001 0.93 --- --- --- ---
model3 −0.077 0.039 −0.005 0.003 0.010 0.68 −0.002 0.041 −0.001 0.76 −0.001 0.91 −0.007 0.35 --- ---
model4 −0.088 0.045 −0.005 0.004 0.010 0.68 −0.003 0.039 −0.001 0.70 −0.001 0.91 −0.008 0.34 5.6e-5 0.68
Women
SSAR1_N C20:0 Arachidic Acid
SSAR1_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.098 0.21 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.097 0.30 −0.002 0.37 0.025 0.59 −0.003 0.18 −0.008 0.051 --- --- --- --- --- ---
model2 −0.092 0.33 −0.002 0.35 0.026 0.57 −0.003 0.21 −0.008 0.08 −0.019 0.61 --- --- --- ---
model3 −0.090 0.36 −0.002 0.33 0.027 0.57 −0.003 0.22 −0.007 0.13 −0.013 0.73 −0.007 0.64 --- ---
model4 −0.105 0.33 −0.002 0.37 0.026 0.58 −0.003 0.21 −0.008 0.12 −0.016 0.69 −0.009 0.59 1e-4 0.74
SSDA1_N C22:0 Docosanoic Acid
SSDA1_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.165 0.022 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.113 0.17 −0.002 0.43 0.030 0.52 −0.003 0.18 −0.009 0.029 --- --- --- --- --- ---
model2 −0.122 0.14 −0.002 0.42 0.031 0.51 −0.003 0.23 −0.008 0.06 −0.030 0.42 --- --- --- ---
model3 −0.121 0.15 −0.002 0.40 0.031 0.50 −0.003 0.24 −0.008 0.10 −0.024 0.54 −0.007 0.65 --- ---
model4 −0.126 0.15 −0.002 0.42 0.032 0.51 −0.003 0.23 −0.008 0.10 −0.027 0.51 −0.008 0.62 1e-4 0.80
SSARA_N C20:4 n-6 Arachidonic Acid (AA)
SSARA_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.147 0.017 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.15 0.038 −0.002 0.32 0.043 0.33 −0.003 0.21 −0.007 0.10 --- --- --- --- --- ---
model2 −0.147 0.042 −0.002 0.30 0.044 0.32 −0.003 0.25 −0.006 0.15 −0.018 0.61 --- --- --- ---
model3 −0.148 0.045 −0.002 0.30 0.044 0.33 −0.003 0.25 −0.006 0.20 −0.016 0.67 −0.003 0.84 --- ---
model4 −0.163 0.040 −0.002 0.34 0.045 0.31 −0.003 0.21 −0.006 0.17 −0.021 0.59 −0.005 0.75 2e-4 0.59
SSDTA_N C22:4 n-6 Docosatetraenoic Acid (DTA)
SSDTA_N (log) Age (yr) Smoking (Y/N) dBP BMI CRP HOMA_IR TG
β p β p β p β p β p β p β p β p
model0 −0.043 0.40 --- --- --- --- --- --- --- --- --- --- --- --- --- ---
model1 −0.093 0.11 −0.003 0.19 0.046 0.31 −0.003 0.23 −0.007 0.11 --- --- --- --- --- ---
model2 −0.090 0.13 −0.003 0.18 0.046 0.31 −0.003 0.25 −0.006 0.14 −0.013 0.72 --- --- --- ---
model3 −0.089 0.14 −0.003 0.18 0.046 0.31 −0.003 0.26 −0.006 0.18 −0.011 0.77 −0.003 0.84 --- ---
model4 −0.129 0.09 −0.003 0.23 0.052 0.26 −0.003 0.18 −0.007 0.13 −0.017 0.66 −0.006 0.69 4e-4 0.37
Open in a new tab

The effects of various combinations of potential confounding factors were adjusted with multivariate models as follows: Model 0: log VO2max ~ log FA (univariate analysis); Model 1: log VO2max ~ log FA + Age + Smoking + dBP + BMI; Model 2: log VO2max ~ log FA + Age + Smoking + dBP + BMI + CRP; Model 3: log VO2max ~ log FA + Age + Smoking + dBP + BMI + CRP + HOMA-IR; Model 4: log VO2max ~ log FA + Age + Smoking + dBP + BMI + CRP + HOMA-IR + TG; Abbreviations: dBP: diastolic Blood Pressure (mmHg); BMI: Body Mass Index (kg/m2); CRP: Plasma C-Reactive Protein (mg/dL); β: coefficient of variable; HOMA_IR: HOmeostatic Model Assessment – Insulin Resistance; TG: Plasma Triglyceride (mg/dL).

4. Discussion

This study shows for the first-time that plasma concentrations of 2 VLSFAs (AR1, DA1), and 2 n-6 PUFAs (AA, DTA) appear to be higher in young and middle-aged general adults with low CVFL, and intriguingly, these associations are sex-dimorphic. The only FA showed negative correlation with VO2max in women is AA and the significance could only be revealed with multivariate analysis after adjusting potential confounders. In contrast, the relationships between VO2max those 4 FAs were seen consistently in men.

The lack of established reference ranges for saturated, trans, monounsaturated and polyunsaturated FA has resulted in the poor interpretability of human research [23]. Most of the researches concerning FAs and cardiovascular health so far have either used frequent food questionnaire to estimate the FAs levels or measured the whole non-esterified fatty acid (NEFA) levels instead of the individual FAs concentrations. Fortunately, there has been an increasing appreciation of the values of measuring circulating FAs to better translate its biomedical relevance [11,24,25,26].

Previous data suggest that plasma levels of FAs and their composition may influence myocardial function. For example, it has been shown that the composition of FAs in chronic heart failure (HF) is important [16]. Specifically, for n-6 PUFAs levels in HF patients, there was a decrease in the content of C18:2n-6 (linoleic acid), increased C22:5n-6 (DTA) and C22:4n-6 (DPA n-6) but no changes in C20:4n-6 (AA). In the current study, these subjects were “apparently healthy” despite of different CVFLs. While we did not find significant difference of n-3 PUFAs among CVFL groups, interestingly, we did observe increased DTA in the low CVFL group, as seen in HF patients. AA also negatively correlated with CVFL in our cohort but there was no difference in the more advanced setting (HF). Whether these individual n-6 PUFAs have any impact on future cardiovascular events merit further mechanistic study.

The potential beneficial role of n-6 PUFAs has been debated [27,28]. Despite the fact that most dietary recommendations agree that SFAs should be at least partially replaced by unsaturated FAs, in particular vegetable PUFAs, the suggestion to increase the intake of PUFAs in general, and n-6 PUFAs in particular, continues to be controversial [27,28,29,30]. n-6 PUFAs, such as arachidonic acid (AA; 20:4n-6), which is the substrate for the synthesis of a variety of proinflammatory and vasoconstrictive molecules, are believed to be proinflammatory [31,32]. On the other hand, despite the concern that n-6 fatty acids increase inflammation, evidence from human studies does not support this view [29]. Nevertheless, a recent meta-analysis reviewed 19 randomized controlled intervention trials with 6461 participants and concluded that there was no evidence that increasing omega-6 fat reduces cardiovascular outcomes other than myocardial infarction [33].

Two VLSFAs, Arachidic acid (AR1, C20:0), Docosanoic acid (DA1, or Behenic acid, C22:0) were inversely correlated with CVFL in men in the current study. Several prospective investigations have, however, documented potentially beneficial associations of this unique group, saturated fats with a chain length of 20 or more carbon atoms, with cardiometabolic disease [26,34,35]. VL SFAs are derived mainly from dietary sources and may also be produced endogenously from the elongation of 18:0 to 20:0, 22:0, and 24:0 [36,37]. VLSFAs, however, exhibit distinct functions when compared with other long-chain SFAs. For example, VLSFAs are known to influence liver homeostasis, retinal function, and anti-inflammatory functions [37]. Given the reported beneficial association with various conditions, it is hard to interpret the opposite associations with CVFL. Indeed, in contrast to prospective results, evidence generated from cross-sectional studies has been somewhat mixed [35]. One possible source of the differences between studies could be the methodology of FAs levels assessment from either food questionnaire or the compartment where (such as in serum whole blood, plasma phospholipids or membrane of blood cells) the measurements were done. For example, some studies reported that erythrocyte C24:0 or C26:0 were consistently correlated with atherogenic lipoprotein profiles or diabetes status [38,39]. Further research is needed to elucidate the mechanisms underlying the observed associations.

The sex dimorphism in cardiovascular risks, including lipid metabolism and insulin resistance has been well documented [40,41,42,43,44]. Moreover, it has also been widely accepted that biological sex is an important modifier of the development of cardiovascular disease [45,46]: sex differences are apparent at every level of cardiovascular physiology from action potential duration and mitochondrial energetics to cardiomyocytes and whole heart contractile function [47]. Premenopausal women have reduced levels of CVD compared with their age-matched counterparts [48]. Additionally, baseline FAs levels, FA disposal, and even the clinical effects of FAs supplementation are different in women and man. In general, women have greater nonoxidative FFA disposal and recycling than men [49,50], and the rate of FFA release into plasma is greater in women than men [51]. Additionally, women in the general population have higher circulating DHA concentrations, but lower circulating EPA and DPA concentrations compared with men and this difference is independent of dietary intake [52,53,54]. Interestingly, a recent meta-analysis including 31 eligible randomized controlled trials with a total of 1848 participants reported that a significant improvement in insulin resistance with n-3 PUFA supplementation longer than 6 weeks was only seen in women but not in men [55]. Similarly, n-3 PUFA supplementation enhances muscle function in older women but not in older men after resistance exercise training [56]. Considering that many known cardiovascular risk factors are sex-dimorphic, and that many aspects of FAs are different in women and men, we thus also further stratified the subjects by gender, and examine the association of CVFL with FA levels separately. Not expectedly, all the associations observed came from men.

Among the 4 FAs of interest, men have higher DTA while the levels other 3 FAs were similar. The absolute concentrations of individual FA per se might not be the explanation of the dimorphism. Another possibility is that greater variation of FAs concentration exists in women and masks its biological relevance. This is probably not the case since the coefficients of variance CV (%) of FAs of women were not greater than those of men. Additionally, it has been reported that plasma FAs concentrations were not affected by menstrual cycle [57]. Limited by the nature of the current study, we could not explain the mechanism for the sex-specific associations of FAs and CVFLs. In addition, even though the molecular basis underlying this sex dimorphism remains to be elucidated, further study may provide important insights for gender-specific prevention or intervention strategy.

The present study has a few limitations. FA levels in the current study were measured only in plasma but not other compartments, such as lipoproteins or plasma phospholipids on membranes of blood cells. Therefore, apart from being used as biomarkers, these levels could not provide the whole pictures of FAs metabolism. Also, as a cross-sectional epidemiologic study, these statistical associations do not necessarily imply any causal relationships, and the mechanisms underlying these observations could not be established. Moreover, generalizability of the results seems difficult because the subsample was not chosen randomly but due to the data availability of both CVFLs and FAs. Importantly, although we adjusted for factors that have previously reported to be associated with CVFL [58,59], we cannot exclude the role of residual or unmeasured confounding in the findings. Further researches are required to investigate factors that may modulate effects of these fatty acids.

5. Conclusions

In conclusion, in young and middle-aged adults with low CVFL, plasma concentrations of 2 VLSFAs (AR1, DA1) and 2 n-6 PUFAs (AA1, DTA) were found to be higher than those of moderate and high CVFL. These associations were seen only in men but not women. Further studies are required to verify the presence of the observed sexual dimorphism in the association of FAs and CVFL, and to investigate the potential impact of these associations on long-term cardiovascular outcome.

Acknowledgments

The authors would like to thank all participants involved in NHANES surveys.

Author Contributions

Conceptualization, P.-L.T.; Data curation, P.-L.T.; Formal analysis, P.-L.T. and C.-J.W.; Investigation, P.-L.T. and C.-J.W.; Methodology, P.-L.T. and C.-J.W.; Supervision, C.-J.W.; Writing – original draft, P.-L.T.; Writing – review & editing, P.-L.T. and C.-J.W.

Funding

This research received no external funding.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  • 1.Sallis J.F., Patterson T.L., Buono M.J., Nader P.R. Relation of cardiovascular fitness and physical activity to cardiovascular disease risk factors in children and adults. Am. J. Epidemiol. 1988;127:933–941. doi: 10.1093/oxfordjournals.aje.a114896. [DOI] [PubMed] [Google Scholar]
  • 2.Blair S.N., Kohl H.W., Paffenbarger R.S., Clark D.G., Cooper K.H., Gibbons L.W. Physical fitness and all-cause mortality. A prospective study of healthy men and women. JAMA. 1989;262:2395–2401. doi: 10.1001/jama.1989.03430170057028. [DOI] [PubMed] [Google Scholar]
  • 3.Carnethon M.R., Gulati M., Greenland P. Prevalence and cardiovascular disease correlates of low cardiorespiratory fitness in adolescents and adults. JAMA. 2005;294:2981–2988. doi: 10.1001/jama.294.23.2981. [DOI] [PubMed] [Google Scholar]
  • 4.Ekelund L.G., Haskell W.L., Johnson J.L., Whaley F.S., Criqui M.H., Sheps D.S. Physical fitness as a predictor of cardiovascular mortality in asymptomatic North American men. The lipid research clinics mortality follow-up study. New Engl. J. Med. 1988;319:1379–1384. doi: 10.1056/NEJM198811243192104. [DOI] [PubMed] [Google Scholar]
  • 5.Kodama S., Saito K., Tanaka S., Maki M., Yachi Y., Asumi M., Sugawara A., Totsuka K., Shimano H., Ohashi Y., et al. Cardiorespiratory fitness as a quantitative predictor of all-cause mortality and cardiovascular events in healthy men and women: A meta-analysis. JAMA. 2009;301:2024–2035. doi: 10.1001/jama.2009.681. [DOI] [PubMed] [Google Scholar]
  • 6.Clausen J.S.R., Marott J.L., Holtermann A., Gyntelberg F., Jensen M.T. Midlife cardiorespiratory fitness and the long-term risk of mortality: 46 years of follow-up. J. Am. Coll. Cardiol. 2018;72:987–995. doi: 10.1016/j.jacc.2018.06.045. [DOI] [PubMed] [Google Scholar]
  • 7.Oliver M.F. Sudden cardiac death: The lost fatty acid hypothesis. QJM Mon. J. Assoc. Physicians. 2006;99:701–709. doi: 10.1093/qjmed/hcl084. [DOI] [PubMed] [Google Scholar]
  • 8.Djousse L., Benkeser D., Arnold A., Kizer J.R., Zieman S.J., Lemaitre R.N., Tracy R.P., Gottdiener J.S., Mozaffarian D., Siscovick D.S., et al. Plasma free fatty acids and risk of heart failure: The cardiovascular health study. Circ. Heart Fail. 2013;6:964–969. doi: 10.1161/CIRCHEARTFAILURE.113.000521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Rider O.J., Holloway C.J., Emmanuel Y., Bloch E., Clarke K., Neubauer S. Increasing plasma free fatty acids in healthy subjects induces aortic distensibility changes seen in obesity. Circ. Cardiovasc. Imaging. 2012;5:367–375. doi: 10.1161/CIRCIMAGING.111.971804. [DOI] [PubMed] [Google Scholar]
  • 10.Zhu N., Jiang W., Wang Y., Wu Y., Chen H., Zhao X. Plasma levels of free fatty acid differ in patients with left ventricular preserved, mid-range, and reduced ejection fraction. BMC Cardiovasc. Disord. 2018;18:104. doi: 10.1186/s12872-018-0850-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Abdelmagid S.A., Clarke S.E., Nielsen D.E., Badawi A., El-Sohemy A., Mutch D.M., Ma D.W. Comprehensive profiling of plasma fatty acid concentrations in young healthy Canadian adults. PLoS ONE. 2015;10:e0116195. doi: 10.1371/journal.pone.0116195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Sibbons C.M., Brenna J.T., Lawrence P., Hoile S.P., Clarke-Harris R., Lillycrop K.A., Burdge G.C. Effect of sex hormones on n-3 polyunsaturated fatty acid biosynthesis in HepG2 cells and in human primary hepatocytes. Prostaglandins Leukot. Essent. Fat. Acids. 2014;90:47–54. doi: 10.1016/j.plefa.2013.12.006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Steffen B.T., Steffen L.M., Tracy R., Siscovick D., Jacobs D., Liu K., He K., Hanson N.Q., Nettleton J.A., Tsai M.Y. Ethnicity, plasma phospholipid fatty acid composition and inflammatory/endothelial activation biomarkers in the Multi-Ethnic Study of Atherosclerosis (MESA) Eur. J. Clin. Nutr. 2012;66:600–605. doi: 10.1038/ejcn.2011.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Kurotani K., Sato M., Ejima Y., Nanri A., Yi S., Pham N.M., Akter S., Poudel-Tandukar K., Kimura Y., Imaizumi K., et al. High levels of stearic acid, palmitoleic acid, and dihomo-gamma-linolenic acid and low levels of linoleic acid in serum cholesterol ester are associated with high insulin resistance. Nutr. Res. 2012;32:669–675. doi: 10.1016/j.nutres.2012.07.004. [DOI] [PubMed] [Google Scholar]
  • 15.Yamagishi K., Folsom A.R., Steffen L.M., Investigators A.S. Plasma fatty acid composition and incident ischemic stroke in middle-aged adults: The Atherosclerosis Risk in Communities (ARIC) Study. Cerebrovasc. Dis. 2013;36:38–46. doi: 10.1159/000351205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Oie E., Ueland T., Dahl C.P., Bohov P., Berge C., Yndestad A., Gullestad L., Aukrust P., Berge R.K. Fatty acid composition in chronic heart failure: Low circulating levels of eicosatetraenoic acid and high levels of vaccenic acid are associated with disease severity and mortality. J. Intern. Med. 2011;270:263–272. doi: 10.1111/j.1365-2796.2011.02384.x. [DOI] [PubMed] [Google Scholar]
  • 17.NHANES 2003–2004. [(accessed on 24 July 2018)]; Available online: https://wwwn.cdc.gov/nchs/nhanes/ContinuousNhanes/Default.aspx?BeginYear=2003.
  • 18.National Health and Examination Survey: Cardiovascular Fitness Procedures Manual. [(accessed on 24 July 2018)]; Available online: https://wwwn.cdc.gov/nchs/data/nhanes/2003-2004/manuals/cv_99-04.pdf.
  • 19.Pate R.R., Pratt M., Blair S.N., Haskell W.L., Macera C.A., Bouchard C., Buchner D., Ettinger W., Heath G.W., King A.C., et al. Physical activity and public health. A recommendation from the centers for disease control and prevention and the American college of sports medicine. JAMA. 1995;273:402–407. doi: 10.1001/jama.1995.03520290054029. [DOI] [PubMed] [Google Scholar]
  • 20.Lagerstedt S.A., Hinrichs D.R., Batt S.M., Magera M.J., Rinaldo P., McConnell J.P. Quantitative determination of plasma c8–c26 total fatty acids for the biochemical diagnosis of nutritional and metabolic disorders. Mol. Genet. Metab. 2001;73:38–45. doi: 10.1006/mgme.2001.3170. [DOI] [PubMed] [Google Scholar]
  • 21.Strath S.J., Holleman R.G., Ronis D.L., Swartz A.M., Richardson C.R. Objective physical activity accumulation in bouts and nonbouts and relation to markers of obesity in US adults. Prev. Chronic Dis. 2008;5:131. [PMC free article] [PubMed] [Google Scholar]
  • 22.Dwoskin L.P., Buxton S.T., Jewell A.L., Crooks P.A. S (-)-nornicotine increases dopamine release in a calcium-dependent manner from superfused rat striatal slices. J. Neurochem. 1993;60:2167–2174. doi: 10.1111/j.1471-4159.1993.tb03502.x. [DOI] [PubMed] [Google Scholar]
  • 23.Baum S.J. A Survey of internists and cardiologists: Are discoveries in fatty acids truly being translated into clinical practice? Prostaglandins Leukot. Essent. Fat. Acids. 2013;88:3–4. doi: 10.1016/j.plefa.2012.05.001. [DOI] [PubMed] [Google Scholar]
  • 24.Adjepong M., Pickens C.A., Jain R., Harris W.S., Annan R.A., Fenton J.I. Association of whole blood n-6 fatty acids with stunting in 2-to-6-year-old Northern Ghanaian children: A cross-sectional study. PLoS ONE. 2018;13:e0193301. doi: 10.1371/journal.pone.0193301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.de Oliveira Otto M.C., Wu J.H., Baylin A., Vaidya D., Rich S.S., Tsai M.Y., Jacobs D.R., Jr., Mozaffarian D. Circulating and dietary omega-3 and omega-6 polyunsaturated fatty acids and incidence of CVD in the Multi-Ethnic Study of Atherosclerosis. J. Am. Heart Assoc. 2013;2:e000506. doi: 10.1161/JAHA.113.000506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Fretts A.M., Mozaffarian D., Siscovick D.S., Djousse L., Heckbert S.R., King I.B., McKnight B., Sitlani C., Sacks F.M., Song X., et al. Plasma phospholipid saturated fatty acids and incident atrial fibrillation: The cardiovascular health study. J. Am. Heart Assoc. 2014;3:e000889. doi: 10.1161/JAHA.114.000889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Nettleton J.A., von Schacky C., Brouwer I.A., Koletzko B. International society for the study of fatty acids and lipids 2016 debate: For science-based dietary guidelines on fats, meta-analysis and systematic reviews are decisive. Ann. Nutr. Metab. 2017;71:26–30. doi: 10.1159/000478794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Sacks F.M., Lichtenstein A.H., Wu J.H.Y., Appel L.J., Creager M.A., Kris-Etherton P.M., Miller M., Rimm E.B., Rudel L.L., Robinson J.G., et al. Dietary fats and cardiovascular disease: A presidential advisory from the American heart association. Circulation. 2017;136:1–23. doi: 10.1161/CIR.0000000000000510. [DOI] [PubMed] [Google Scholar]
  • 29.Maki K.C., Eren F., Cassens M.E., Dicklin M.R., Davidson M.H. Omega-6 polyunsaturated fatty acids and cardiometabolic health: Current evidence, controversies, and research gaps. Adv. Nutr. 2018 doi: 10.1093/advances/nmy038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.U.S. Department of Health and Human Services and U.S. Department of Agriculture . 2015–2020 Dietary Guidelines for Americans. 8th ed. Skyhorse Publishing; New York, NY, USA: 2015. [Google Scholar]
  • 31.Harris W.S., Mozaffarian D., Rimm E., Kris-Etherton P., Rudel L.L., Appel L.J., Engler M.M., Engler M.B., Sacks F. Omega-6 fatty acids and risk for cardiovascular disease: A science advisory from the American heart association nutrition subcommittee of the council on nutrition, physical activity, and metabolism; council on cardiovascular nursing; and council on epidemiology and prevention. Circulation. 2009;119:902–907. doi: 10.1161/CIRCULATIONAHA.108.191627. [DOI] [PubMed] [Google Scholar]
  • 32.Node K., Huo Y., Ruan X., Yang B., Spiecker M., Ley K., Zeldin D.C., Liao J.K. Anti-inflammatory properties of cytochrome P450 epoxygenase-derived eicosanoids. Science. 1999;285:1276–1279. doi: 10.1126/science.285.5431.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Hooper L., Al-Khudairy L., Abdelhamid A.S., Rees K., Brainard J.S., Brown T.J., Ajabnoor S.M., O’Brien A.T., Winstanley L.E., Donaldson D.H., et al. Omega-6 fats for the primary and secondary prevention of cardiovascular disease. Cochrane Database Syst. Rev. 2018;7:CD011094. doi: 10.1002/14651858.CD011094.pub3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Forouhi N.G., Koulman A., Sharp S.J., Imamura F., Kroger J., Schulze M.B., Crowe F.L., Huerta J.M., Guevara M., Beulens J.W., et al. Differences in the prospective association between individual plasma phospholipid saturated fatty acids and incident type 2 diabetes: The EPIC-InterAct case-cohort study. Lancet Diabetes Endocrinol. 2014;2:810–818. doi: 10.1016/S2213-8587(14)70146-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Malik V.S., Chiuve S.E., Campos H., Rimm E.B., Mozaffarian D., Hu F.B., Sun Q. Circulating very-long-chain saturated fatty acids and incident coronary heart disease in US men and women. Circulation. 2015;132:260–268. doi: 10.1161/CIRCULATIONAHA.114.014911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Jakobsson A., Westerberg R., Jacobsson A. Fatty acid elongases in mammals: Their regulation and roles in metabolism. Prog. Lipid Res. 2006;45:237–249. doi: 10.1016/j.plipres.2006.01.004. [DOI] [PubMed] [Google Scholar]
  • 37.Kihara A. Very long-chain fatty acids: Elongation, physiology and related disorders. J. Biochem. 2012;152:387–395. doi: 10.1093/jb/mvs105. [DOI] [PubMed] [Google Scholar]
  • 38.Antoku Y., Tsukamoto K., Miyoshi Y., Nagino H., Anezaki M., Suwa K., Narabe Y. Correlations of elevated levels of hexacosanoate in erythrocyte membranes with risk factors for atherosclerosis. Atherosclerosis. 2000;153:169–173. doi: 10.1016/S0021-9150(00)00393-2. [DOI] [PubMed] [Google Scholar]
  • 39.Matsumori R., Miyazaki T., Shimada K., Kume A., Kitamura Y., Oshida K., Yanagisawa N., Kiyanagi T., Hiki M., Fukao K., et al. High levels of very long-chain saturated fatty acid in erythrocytes correlates with atherogenic lipoprotein profiles in subjects with metabolic syndrome. Diabetes Res. Clin. Pract. 2013;99:12–18. doi: 10.1016/j.diabres.2012.10.025. [DOI] [PubMed] [Google Scholar]
  • 40.Alfredsson J., Green J.B., Stevens S.R., Reed S.D., Armstrong P.W., Angelyn Bethel M., Engel S.S., McGuire D.K., Van de Werf F., Hramiak I., et al. Sex differences in management and outcomes of patients with type 2 diabetes and cardiovascular disease: A report from TECOS. Diabetes Obes. Metab. 2018 doi: 10.1111/dom.13377. [DOI] [PubMed] [Google Scholar]
  • 41.Cenko E., Yoon J., Kedev S., Stankovic G., Vasiljevic Z., Krljanac G., Kalpak O., Ricci B., Milicic D., Manfrini O., et al. Sex differences in outcomes after STEMI: Effect modification by treatment strategy and age. JAMA Intern. Med. 2018;178:632–639. doi: 10.1001/jamainternmed.2018.0514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Palmisano B.T., Zhu L., Eckel R.H., Stafford J.M. Sex differences in lipid and lipoprotein metabolism. Mol. Metab. 2018;15:45–55. doi: 10.1016/j.molmet.2018.05.008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Santilli F., D’Ardes D., Guagnano M.T., Davi G. Metabolic syndrome: Sex-related cardiovascular risk and therapeutic approach. Curr. Med. Chem. 2017;24:2602–2627. doi: 10.2174/0929867324666170710121145. [DOI] [PubMed] [Google Scholar]
  • 44.Suthahar N., Meijers W.C., Ho J.E., Gansevoort R.T., Voors A.A., van der Meer P., Bakker S.J.L., Heymans S., van Empel V., Schroen B., et al. Sex-specific associations of obesity and N-terminal pro-B-type natriuretic peptide levels in the general population. Eur. J. Heart Fail. 2018;20:1205–1214. doi: 10.1002/ejhf.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Maric-Bilkan C., Arnold A.P., Taylor D.A., Dwinell M., Howlett S.E., Wenger N., Reckelhoff J.F., Sandberg K., Churchill G., Levin E., et al. Report of the national heart, lung, and blood institute working group on sex differences research in cardiovascular disease: Scientific questions and challenges. Hypertension. 2016;67:802–807. doi: 10.1161/HYPERTENSIONAHA.115.06967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Abubakar I.I., Tillmann T., Banerjee A. Global, regional, and national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990–2013: A systematic analysis for the global burden of disease study 2013. Lancet. 2015;385:117–171. doi: 10.1016/S0140-6736(14)61682-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.Blenck C.L., Harvey P.A., Reckelhoff J.F., Leinwand L.A. The importance of biological sex and estrogen in rodent models of cardiovascular health and disease. Circ. Res. 2016;118:1294–1312. doi: 10.1161/CIRCRESAHA.116.307509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48.Huxley V.H. Sex and the cardiovascular system: The intriguing tale of how women and men regulate cardiovascular function differently. Adv. Physiol. Educ. 2007;31:17–22. doi: 10.1152/advan.00099.2006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 49.Koutsari C., Basu R., Rizza R.A., Nair K.S., Khosla S., Jensen M.D. Nonoxidative free fatty acid disposal is greater in young women than men. J. Clin. Endocrinol. Metab. 2011;96:541–547. doi: 10.1210/jc.2010-1651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 50.Shadid S., Koutsari C., Jensen M.D. Direct free fatty acid uptake into human adipocytes in vivo: Relation to body fat distribution. Diabetes. 2007;56:1369–1375. doi: 10.2337/db06-1680. [DOI] [PubMed] [Google Scholar]
  • 51.Adler-Wailes D.C., Periwal V., Ali A.H., Brady S.M., McDuffie J.R., Uwaifo G.I., Tanofsky-Kraff M., Salaita C.G., Hubbard V.S., Reynolds J.C., et al. Sex-associated differences in free fatty acid flux of obese adolescents. J. Clin. Endocrinol. Metab. 2013;98:1676–1684. doi: 10.1210/jc.2012-3817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 52.Bakewell L., Burdge G.C., Calder P.C. Polyunsaturated fatty acid concentrations in young men and women consuming their habitual diets. Br. J. Nutr. 2006;96:93–99. doi: 10.1079/BJN20061801. [DOI] [PubMed] [Google Scholar]
  • 53.Crowe F.L., Skeaff C.M., Green T.J., Gray A.R. Serum n-3 long-chain PUFA differ by sex and age in a population-based survey of New Zealand adolescents and adults. Br. J. Nutr. 2008;99:168–174. doi: 10.1017/S000711450779387X. [DOI] [PubMed] [Google Scholar]
  • 54.Giltay E.J., Gooren L.J., Toorians A.W., Katan M.B., Zock P.L. Docosahexaenoic acid concentrations are higher in women than in men because of estrogenic effects. Am. J. Clin. Nutr. 2004;80:1167–1174. doi: 10.1093/ajcn/80.5.1167. [DOI] [PubMed] [Google Scholar]
  • 55.Abbott K.A., Burrows T.L., Thota R.N., Acharya S., Garg M.L. Do omega-3 PUFAs affect insulin resistance in a sex-specific manner? A systematic review and meta-analysis of randomized controlled trials. Am. J. Clin. Nutr. 2016;104:1470–1484. doi: 10.3945/ajcn.116.138172. [DOI] [PubMed] [Google Scholar]
  • 56.Da Boit M., Sibson R., Sivasubramaniam S., Meakin J.R., Greig C.A., Aspden R.M., Thies F., Jeromson S., Hamilton D.L., Speakman J.R., et al. Sex differences in the effect of fish-oil supplementation on the adaptive response to resistance exercise training in older people: A randomized controlled trial. Am. J. Clin. Nutr. 2017;105:151–158. doi: 10.3945/ajcn.116.140780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 57.Magkos F., Patterson B.W., Mittendorfer B. No effect of menstrual cycle phase on basal very-low-density lipoprotein triglyceride and apolipoprotein B-100 kinetics. Am. J. Physiol. Endocrinol. Metab. 2006;291:1243–1249. doi: 10.1152/ajpendo.00246.2006. [DOI] [PubMed] [Google Scholar]
  • 58.Kuo H.K., Yen C.J., Chen J.H., Yu Y.H., Bean J.F. Association of cardiorespiratory fitness and levels of C-reactive protein: Data from the national health and nutrition examination survey 1999–2002. Int. J. Cardiol. 2007;114:28–33. doi: 10.1016/j.ijcard.2005.11.110. [DOI] [PubMed] [Google Scholar]
  • 59.Wang C.Y., Haskell W.L., Farrell S.W., Lamonte M.J., Blair S.N., Curtin L.R., Hughes J.P., Burt V.L. Cardiorespiratory fitness levels among US adults 20–49 years of age: Findings from the 1999–2004 national health and nutrition examination survey. Am. J. Epidemiol. 2010;171:426–435. doi: 10.1093/aje/kwp412. [DOI] [PubMed] [Google Scholar]

Articles from Nutrients are provided here courtesy of Multidisciplinary Digital Publishing Institute (MDPI)

RESOURCES