Rinnakkaistallenteet Terveystieteiden tiedekunta
2018
Intake of Fatty Fish Alters the Size and the Concentration of Lipid Components of HDL Particles and Camelina Sativa Oil Decreases IDL Particle
Concentration in Subjects with Impaired Glucose Metabolism
Manninen, Suvi M
Wiley
Tieteelliset aikakauslehtiartikkelit
© WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim All rights reserved
http://dx.doi.org/10.1002/mnfr.201701042
https://erepo.uef.fi/handle/123456789/6666
Downloaded from University of Eastern Finland's eRepository
Research Article
Intake of fish alters the size and composition of HDL particles and camelina sativa oil decreases IDL particle concentration in subjects with impaired glucose metabolism
Suvi M. Manninen1, Maria A. Lankinen1, Vanessa D. de Mello1, David E. Laaksonen2,3, Ursula S.
Schwab1,2, Arja T. Erkkilä1
1Institute of Public Health and Clinical Nutrition, Faculty of Health Sciences, University of Eastern Finland, Kuopio, Finland
2Institute of Clinical medicine, Internal Medicine, Kuopio University Hospital, Kuopio, Finland
3Institute of Biomedicine, Physiology, University of Eastern Finland, Kuopio, Finland
Keywords: α-linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, lipoprotein subclasses, n-3 fatty acids
Abbreviations: ALA, alpha-linolenic acid; CHD, coronary heart disease; CVD, cardiovascular disease; DHA, docosahexaenoic acid; EPA, eicosapentaenoic acid; NMR, nuclear magnetic resonance.
Clinical Trial Registry: Study is registered in Clinicaltrials.gov (NCT01768429).
Correspondence: Suvi M. Manninen, M.Sc.
Address:
University of Eastern Finland, Kuopio Campus School of Medicine
Institute of Public Health and Clinical Nutrition P.O.Box 1627
70211 Kuopio, Finland email: suvi.manninen@uef.fi phone: +358 40 576 1442
Scope: Intake of long-chain n-3 PUFAs affects the lipoprotein subclass profile, whereas the effect 1
of shorter chain n-3 PUFAs remains unclear. We investigated the effect of fish and camelina sativa 2
oil (CSO) intakes on lipoprotein subclasses.
3
Methods and results: Altogether 79 volunteers with impaired glucose metabolism were randomly 4
assigned to CSO, fatty fish (FF), lean fish (LF) or control group for 12 weeks. Nuclear magnetic 5
resonance spectroscopy was used to determine lipoprotein subclasses and their lipid components.
6
The average HDL particle size increased in the FF group (overall p=0.032) as compared with the 7
control group. Serum concentrations of cholesterol in HDL and HDL2 (overall p=0.024 and 8
p=0.021, respectively) and total lipids and phospholipids in large HDL particles (overall p=0.012 9
and p=0.019, respectively) increased in the FF group, differing significantly from the LF group. The 10
concentration of intermediate-density lipoprotein (IDL) particles decreased in the CSO group 11
(overall p=0.033) as compared with the LF group.
12
Conclusion: Our study suggests that FF intake causes a shift towards larger HDL particles and 13
increases the concentration of lipid components in HDL, which may be associated with the 14
antiatherogenic properties of HDL. Furthermore, CSO intake decreases IDL particle concentration.
15
These changes may favorably affect cardiovascular risk.
16 17
1 Introduction 18
Intake of long-chain n-3 polyunsaturated fatty acids (PUFA) is known to have cardiovascular 19
benefits [1,2]. The mechanisms behind the beneficial effects of n-3 PUFA on the risk factors of 20
cardiovascular disease (CVD) are not fully understood, but their effects on lipoprotein metabolism 21
are potential contributing factors [3,4]. The main long-chain n-3 PUFAs are eicosapentaenoic acid 22
(EPA) and docosahexaenoic acid (DHA), which are primarily derived from fish [1]. Alpha-linolenic 23
acid (ALA) is a shorter chain n-3 fatty acid from plant sources and a precursor for EPA and DHA 24
synthesis [5]. However, the degree of conversion of ALA to EPA and DHA varies extensively. The 25
effects of ALA intake on CVD risk factors remains unclear, but it may decrease the risk of coronary 26
heart disease (CHD) when long-chain n-3 PUFA intake is low [6].
27 28
High serum concentrations of LDL cholesterol and low serum concentrations of HDL cholesterol 29
are known risk factors for CVD [7]. However, HDL and LDL are not homogeneous groups of 30
particles, but have subfractions differing in size and density as well as in lipid and apolipoprotein 31
compositions [8]. These subfractions have different impact on the risk of CVD; large HDL particles 32
may be more atheroprotective than small HDL particles [9-11].Conversely, small LDL particles 33
may be more atherogenic than large LDL particles [12,13].
34 35
There are only a few intervention studies that have examined the effect of fish intake on lipoprotein 36
subclasses, especially in individuals at high risk of developing CVD [14-21]. In these studies, fish 37
intake has been reported to decrease the size and concentrations VLDL particles and increase the 38
size and concentrations of HDL particles and lipid components in the particles as compared with 39
control diets. Furthermore, associations of higher intake of dietary n-3 PUFA with particle 40
concentrations and size of HDL and VLDL have been found in cross-sectional studies [22-24].
41
Studies suggest that especially fatty fish intake favorably affects the lipoprotein subclass profile 42
[15-17], but beneficial effects have been found also during lean fish intake [18-20]. Earlier 43
intervention studies investigating the effects of ALA intake on lipoprotein characteristics have 44
reported mixed results [25-29], mostly no effects [25-27].
45 46
To this end, the aim of this study was to investigate how intakes of fatty and lean fish and camelina 47
sativa oil (CSO), a source of ALA, affect lipoprotein subclasses in subjects with impaired glucose 48
metabolism.
49 50 51
2 Materials and methods 52
53
2.1 Subjects 54
Altogether 96 Caucasian volunteers aged 40 to 75 years were recruited in Kuopio area to the study 55
(Figure 1)via advertisements in newspapers, noticeboards and intranet of the university and from 56
previous clinical trials at our Department. The main inclusion criterion was fasting plasma glucose 57
concentration 5.6-6.9 mmol/l. The 2-h glucose concentration in the oral glucose tolerance test 58
(OGTT) had to be < 11.0 mmol/l. Other inclusion criteria were: BMI 25-36 kg/m2, concentrations 59
of fasting serum total cholesterol < 7 mmol/l, LDL-cholesterol < 5.0 mmol/l and total triglycerides 60
< 4.0 mmol/l. The main exclusion criteria included any chronic disease, a condition hampering the 61
ability to follow the dietary intervention protocol, alcohol abuse (> 40 g/d), and weight loss of >5 % 62
during the preceding 6 months. Altogether 79 subjects completed the study (Figure 1). There were 63
no differences in age, sex, fasting plasma glucose or serum lipids between the dropouts and the 64
subjects who completed the study. The power calculation was based on differences in DHA in 65
plasma phospholipids, a valid biomarker of dietary intake [30] (n = 18 per group, total n = 72, 66
difference of 1.2 mol %, when alpha < 0.05 and beta > 0.9), because at the time of the onset of this 67
study there was not enough data on lipoprotein particle size and composition to be used for power 68
calculations.
69 70
2.2 Study design 71
Recruitment for AlfaFish study started in autumn 2012 and the intervention was completed in June 72
2014. In the run-in phase of the intervention, the subjects followed their conventional diet for four 73
weeks and were not allowed to use any oil supplements or products enriched in plant stanols or 74
sterols. After the run-in period, the subjects were randomly assigned into CSO, lean fish (LF), fatty 75
fish (FF) or control group for 12 weeks. Randomization wasstratified by sex and use of statins and 76
performed by the study nurse. The subjects visited the study clinic at 0 and 12 weeks and the blood 77
samples were drawn after 10-hour overnight fasting. The serum lipid profile was analyzed by the 78
methodology in use at the UEF and the Eastern Finland Laboratory Center (ISLAB). Details of the 79
methodology of lipid analyses have been described earlier [15]. Physical activity, alcohol intake, 80
smoking, body weight and use of medication known to affect the measures of lipid metabolism 81
were kept constant during the study.
82 83
The study diets were isocaloric and current nutrient recommendations [31] were followed except for 84
fish and ALA intakes. Subjects in the fish groups were instructed to consume 4 meals of fish (100–
85
150 g per meal) per week as lunch or dinner: in the FF group e.g. salmon, rainbow trout, Baltic 86
herring, vendace, whitefish and mackerel to provide around 1 g of EPA+DHA per day and in the LF 87
group e.g. tuna, pike, perch, pike-perch, saithe and cod. The tuna fish consumed by the subjects was 88
canned tuna (data not shown) and it did not have the same content of EPA and DHA as fresh tuna 89
(fineli.fi). The CSO group ingested camelina sativa oil (27 g) in order to get 10 g of ALA per day.
90
The control and CSO groups were allowed to eat 1 fish meal per week and were instructed to 91
consume lean meat and chicken. Compliance with the study diets was assessed with the fatty acid 92
composition of plasma phospholipids determined by gas chromatography as previously described 93
[32], with food records and with daily consumption records: in the fish groups regarding the intake 94
of fish (number of meals and type of fish) and in the CSO group regarding the intake of CSO.
95 96
The study was approved by the Ethical committee of the Hospital District of Northern Savo 97
(55/2012). The subjects received both oral and written information andgave their written informed 98
consent.
99 100
2.3 Lipoprotein subclass and serum lipid analysis by NMR spectroscopy 101
Lipoprotein particle concentration and size were measured from fasting serum samples using high- 102
throughput proton NMR spectroscopy.The details of the methodology have been described 103
previously [33,34]. The lipoprotein subclasses were defined by particle diameter as follows:
104
chylomicrons and largest VLDL particles (average particle diameter at least 75 nm); five different 105
VLDL subclasses: very large VLDL (average particle diameter of 64.0 nm), large VLDL (53.6 nm), 106
medium VLDL (44.5 nm), small VLDL (36.8 nm), and very small VLDL (31.3 nm); IDL (28.6 107
nm); three LDL subclasses: large LDL (25.5 nm), medium LDL (23.0 nm), and small LDL (18.7 108
nm); and four HDL subclasses: very large HDL (14.3 nm), large HDL (12.1 nm), medium HDL 109
(10.9 nm), and small HDL (8.7 nm). The following components of the lipoprotein particles were 110
quantified: phospholipids, triglycerides, cholesterol, free cholesterol, and cholesterol esters. The 111
mean size for VLDL, LDL and HDL particles were calculated by weighting the corresponding 112
subclass diameters with their particle concentrations. Serum lipid extract analyses were performed 113
as described earlier [33,34].
114 115
2.4 Statistical analyses 116
Statistical analyses were performed using the IBM SPSS statistical software (v. 21, IBM Corp., 117
Armonk, NY). The normality of the variables was tested with the Kolmogorov-Smirnov test. The 118
variables with abnormal distribution were normalized with logarithmic transformation. A non- 119
parametric test was used if normal distribution was not achieved with transformation. The paired 120
samples t-test or Wilcoxon signed ranks test was used to compare baseline and endpoint values 121
within the groups. Analysis of covariance (ANCOVA), followed by multiple comparisons test with 122
Bonferroni correction was used to test the changes in lipoprotein particles during intervention.
123
Analyses of covariance were performed using fold changes. Fold changes were calculated by 124
dividing the endpoint values of the variable by their baseline values. Sex, age, use of statins and 125
baseline values were included in the ANCOVA models. Spearman rank correlation was used to 126
calculate correlation coefficients for the baseline associations. P-value < 0.05 was considered as 127
statistically significant. R Project for Statistical Computing version 3.2.2 was used to calculate 128
Benjamini-Hochberg false discovery rate (FDR) to adjust results for multiple comparisons. None of 129
the results remained statistically significant after correction (FDR-p < 0.05).
130 131
3 Results 132
Characteristics of the participants and compliance 133
Mean (± SD) age of the study subjects was 58.9 ± 6.5 years. There were no differences in baseline 134
characteristics among the study groups (Table 1). Reported physical activity of the subjects did not 135
change during the study [35]. The average number of fish meals per week was 4.4 ± 0.4 in the FF, 136
4.3 ± 0.5 in the LF, 0.9 ± 0.4 in the CSO and 0.9 ± 0.4 in the control group during the study. The 137
camelina oil consumption was 25.7 ± 2.7 g per day in the CSO group. Dietary intakes have been 138
reported in detail elsewhere [35]. The proportion of ALA (18:3n-3) in plasma phospholipids 139
increased in CSO group (p < 0.001) and differed significantly from the other groups. Furthermore, 140
the proportion of EPA (20:5n-3) increased (p < 0.001) in the FF group as compared with the LF and 141
control groups and the proportion of DHA (22:6n-3) increased in the FF group (p < 0.001) as 142
compared with the CSO and control groups.
143 144
The effects of fatty fish intake on lipoprotein particles 145
Mean HDL particle diameter (overall difference among the groups p=0.032) increased in the FF 146
group as compared with the control group (Figure 2, Supporting Information Table S1). The 147
changes in the mean particle size for VLDL and LDL did not differ among the groups (Table 2).
148
Serum concentrations of cholesterol in HDL and HDL2 (overall differences among the groups 149
p=0.024 and p=0.021, respectively) and total lipids and phospholipids in large HDL particles 150
(overall differences among the groups p=0.012 and p=0.019, respectively) increased in the FF 151
group, differing significantly from the LF group (Figure 2, Supporting Information Table S1).
152
Furthermore, serum concentration of phospholipids in very large HDL particles increased in the FF 153
group as compared with the LF group when adjusted for baseline values (overall difference among 154
the groups p=0.048). The significance level also remained after further adjustment for sex, age and 155
use of statins (p=0.050).
156 157
The effects of ALA intake on lipoprotein particles and serum lipids 158
Serum concentration of IDL particles decreased in the CSO group (overall difference among the 159
groups p=0.033), differing significantly from the LF group (p=0.043) (Table 2). Furthermore, serum 160
concentration of total lipids in IDL particles and serum concentration of total cholesterol decreased 161
in the CSO group when adjusted for baseline values (overall difference among the groups p=0.030, 162
p=0.042, respectively) but after further adjustment for sex, age and use of statins there were no 163
significant differences among the groups in the Bonferroni-corrected pairwise comparison. A 164
decrease was also observed in the concentration of esterified cholesterol in the CSO group (overall 165
difference among the groups p=0.020).
166
Correlations of DHA and n-3 PUFA and habitual fish consumption with HDL particles 167
Serum concentrations of DHA and total n-3 PUFA correlated with serum concentrations of 168
triglycerides in very large (r=0.293, p=0.009; r=0.352, p=0.001, respectively), large (r=0.287, 169
p=0.01; r=0.228, p=0.043, respectively), medium (r=0.226, p=0.045; r=0.298, p=0.008, 170
respectively) and total HDL particles (r=0.321, p=0.004; r=0.353, p=0.001, respectively) at the 171
baseline (Supporting Information Table S2). Furthermore, DHA and total n-3 PUFA correlated with 172
the concentration and lipid components of medium HDL particles at the baseline. Habitual fish 173
consumption was positively associated with average particle size of HDL, concentrations of very 174
large and large HDL particles and their lipid components at the baseline (Supporting Information 175
Table S2).
176 177
4 Discussion 178
In this study we investigated the effect of fish and ALA intake on the size and composition of 179
lipoprotein particles. We showed that intake of fatty fish 4 times a week significantly increased the 180
average particle size of HDL which is consistent with our earlier studies [15,16]. In addition to our 181
studies there are only three previous trials investigating the effect of fish intake using NMR 182
lipoprotein data. In these studies, high fish intake had no significant effect on HDL particle size 183
[14,20,21]. However, these studies had small samples sizes and in two of these studies the fish 184
consumed was either partly [14] or entirely lean fish [20]. In the present study, an increase in HDL 185
particle size was found only in the FF group whereas in the other groups, particle size decreased, 186
although non-significantly (Figure 2). This may be due to the high DHA content of fatty fish, which 187
has been reported to increase HDL particle size [36].
188 189
Recent studies on the effects of n-3 fatty acids on lipoproteins have focused on the use of 190
supplements, mostly fish oil supplements. Earlier trials using fish oil, EPA or DHA supplements 191
have reported an increase in HDL particle size [37] and an increase in large HDL particles [37,38]
192
compared with control treatments. Studies have also found that n-3 PUFA supplements result in 193
lower concentrations and smaller size of VLDL particles [37-40] and decreased concentrations of 194
IDL and LDL particles [41]. However, these results are not fully comparable with studies 195
investigating dietary intakes of n-3 PUFA since doses of n-3 PUFA in most of the studies using 196
supplements are greater than from dietary sources [42]. Furthermore, there are other nutrients in fish 197
not found in fish oil that may have an atheroprotective effect [43].
198 199
In addition to particle size, HDL lipid content has an essential part in the atheroprotective functions 200
of HDL [44]. The HDL lipidome favorably affect cholesterol efflux, inflammation, oxidative 201
damage and potentially also antithrombotic and vasodilatory activities. We found, that the 202
concentration of phospholipids in very large and large HDL particles increased in the FF group.
203
These changes may be explained by the observed increase in HDL particle size. We have previously 204
shown that the concentration of phospholipids in large HDL particles increases within the tertile 205
with the greatest increase in fish intake [15]. Earlier studies have proposed an inverse relationship 206
between phospholipid content of HDL and CHD [45] and vascular stiffness [46]. Moreover, HDL 207
phospholipids affect cholesterol efflux capacity [47,48] and possibly contribute to the anti- 208
inflammatory activities of HDL [49].
209 210
The concentration of total cholesterol in HDL increased in the FF group. Similar findings have 211
previously been reported in normolipidemic to mildly hyperlipidemic subjects [50] and in our 212
previous study in patients with CHD [16]. Moreover, we found an increase in the concentration of 213
total cholesterol in HDL2. Similarly, Lindqvist et al. [17] found an increase in HDL2 cholesterol 214
after intake of herring 5 times a week for 6 weeks. However, we did not observe change in the 215
HDL2 cholesterol concentration in the LF group in contrast with earlier studies [18,19].
216
217
Our secondary analyses of the baseline associations of habitual fish consumption and HDL particles 218
are in line with the changes found in HDL particles during the intervention. The subjects consumed 219
mainly fatty fish or equally fatty and lean fish before the intervention (data not shown). This further 220
confirms our results that fatty fish intake has favorable effects on the size and composition of HDL 221
particles.
222 223
The effects of marine n-3 PUFA on CVD risk factors have been widely investigated, whereas the 224
knowledge of the effects of ALA intake on CVD is limited [51]. In this study, we found a decrease 225
in the serum concentration of IDL particles in the CSO group. There are only a few intervention 226
studies investigating the effects of ALA on lipoprotein subclass profile [25-29], and to our 227
knowledge there is only one previous study using NMR data [29]. In that study margarines 228
containing sunflower oil, olive oil and rapeseed oil were used in order to get 1.1 % of energy from 229
ALA. There was no significant change in the IDL particle concentration in that study. However, 230
increased ALA intake decreased total and small VLDL particle concentrations as compared with the 231
control diet. It has been shown that high concentrations of VLDL and IDL cholesterol are 232
associated with the increased risk of CHD [52]. Consequently, our results suggest that ALA intake 233
may decrease the risk of CVD, but further studies are needed to investigate the potential 234
mechanisms.
235 236
In previous studies, impaired glucose metabolism has been observed to have an adverse effect on 237
the lipoprotein particle profile [53,54]. VLDL particles have been larger and concentrations of 238
larger VLDL particles higher. Furthermore, a shift in the concentrations of LDL and HDL particles 239
towards smaller particles and decreased concentrations of large LDL and HDL particles have been 240
reported in these studies. Impaired glucose metabolism may also affect also the lipid components of 241
lipoproteins [54]. The concentration of lipids has been found to increase in VLDL particles and 242
decrease in HDL particles. The effects of diets high in fatty fish and ALA on lipoprotein subclasses 243
seen in these overweight subjects with impaired glucose metabolism may therefore not apply to lean 244
individuals with normal glucose metabolism.
245 246
The strength of the current study is good compliance with the study diets. Furthermore, the sample 247
size of this study is comparable with earlier studies [16,17]. However, further studies are needed to 248
investigate the long-term effects of fish and ALA intake on lipoprotein particles. There are also 249
some limitations to consider in our study. Power calculations were based on differences in DHA in 250
plasma phospholipids, and it is possible that there was not enough power to see all changes in these 251
secondary outcome variables. Moreover, Benjamini-Hochberg false discovery rate was also used to 252
adjust results for multiple comparisons, but after using this conservative adjustment our results were 253
no longer statistically significant. Furthermore, possible confounding due to medication, alcohol 254
consumption and physical activity should be considered since statin [55] and alcohol use [56,57]
255
and exercise [58] have been shown to affect lipoprotein particles. However, the changes in the size 256
and composition of HDL and in the concentration of IDL particles were independent of statin use in 257
our study. Adjustment for alcohol use had no effect on the results either (data not shown).
258
Furthermore, the subjects were asked to keep their physical activity constant and according to the 259
reports from the subjects, there were no significant changes in physical activity during the study.
260 261
In conclusion, fatty fish intake 4 times a week alters the size and composition of HDL towards 262
larger and lipid-rich particles. These changes may be associated with the atheroprotective properties 263
of HDL and therefore partly explain the beneficial effects of fish consumption on the prevention of 264
CVD. Furthermore, ALA intake of 10 g per day decreases IDL particle concentration and may 265
therefore have a favorable effect on the risk of CVD.
266
Author contributions
U.S.S and A.T.E are the principal investigators in the Alpha-fish –study. S.M.M. analyzed the data and wrote the article with the help of U.S.S., A.T.E. and M.A.L. who planned and conducted the study together with V.D.M. D.E.L. had the medical charge of the study.
Acknowledgements
The authors thank Ms. Erja Kinnunen and Mr. Tuomas Onnukka for the technical assistance during the study.
This study was supported by Yrjö Jahnsson Foundation, Diabetes Research Foundation, Juho Vainio Foundation, Finnish Cultural Foundation, North Savo Regional fund, State Research
Funding (VTR) of Kuopio University Hospital, Spearhead funding by University of Eastern Finland and Finnish Cultural Foundation. Suomen Kasviöljyt Ltd, Kesko Ltd and Bunge Finland Ltd
provided oil and fat spreads for the study participants.
Conflict of interest statement
None of the authors have a conflict of interest.
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Figure legends
Figure 1. Flow chart of the study.
CSO, camelina sativa oil; OGTT, oral glucose tolerance test.
Figure 2. Changes in mean values (12 wk – 0 wk) for the average diameter of HDL particles (A), serum concentration of total lipids in large HDL (B), serum concentration of total cholesterol in HDL (C), serum concentration of total cholesterol in HDL2 (D), serum concentration of
phospholipids in very large (E) and large HDL (F). Changes among the groups were tested using analysis of covariance adjusted for age, sex, use of statins and baseline values followed by Bonferroni’s post hoc tests. The p-value in the box represents the overall difference among the groups. CSO, camelina sativa oil.
Table 1. Characteristics of the subjects at the baseline of the intervention CSO
(n=18)
Fatty fish (n=20)
Lean fish (n=21)
Control (n=20)
p value a)
Age 58.0 ± 5.6 59.0 ± 6.1 58.1 ± 7.8 60.6 ± 6.2 0.590
Sex, female/male, n/n 10/8 10/10 10/11 9/11 0.928
BMI (kg/m2) 28.7 ± 2.2 29.3 ± 2.0 29.6 ± 3.0 29.3 ± 2.6 0.736
Serum total cholesterol (mmol/l) 5.3 ± 1.0 5.1 ± 1.1 5.4 ± 1.1 5.3 ± 0.9 0.897 LDL cholesterol (mmol/l) 3.2 ± 0.9 3.0 ± 0.9 3.3 ± 0.8 3.2 ± 0.9 0.746 HDL cholesterol (mmol/l) 1.3 ± 0.3 1.4 ± 0.4 1.4 ± 0.5 1.3 ± 0.3 0.463 Triglycerides (mmol/l) b) 1.6 ± 0.6 1.6 ± 0.8 1.3 ± 0.5 1.6 ± 0.7 0.426 Systolic blood pressure (mmHg) 126 ± 12 131 ± 13 129 ± 10 133 ± 11 0.411 Diastolic blood pressure (mmHg) 85 ± 7 85 ± 7 83 ± 8 86 ± 5 0.663 Fasting plasma glucose (mmol/l) b) 6.1 ± 0.4 5.9 ± 0.4 6.1 ± 0.4 6.1 ± 0.6 0.589
Use of statins, n 4 5 4 5 0.966
Habitual fish consumption (meals/week) b) 1.9 ± 0.9 1.9 ± 0.7 2.0 ± 1.0 1.6 ± 0.9 c) 0.580 Values are means ± SD unless otherwise indicated. CSO, camelina sativa oil.
a) p -values were determined by analysis of variance or 2 test.
b) Variables were log-transformed.
c) n=18
CSO (n=18) Fatty fish (n=20) Lean fish (n=21) Control (n=20) p values ANCOVA
0 wk 12 wk 0 wk 12 wk 0 wk 12 wk 0 wk 12 wk Model 1 Model 2 Model 3
Chylomicrons and largest VLDL particles (nmol/l) 0.13 ± 0.10 0.09 ± 0.08 0.16 ± 0.15 0.11 ± 0.07 0.11 ± 0.07 0.14 ± 0.21 0.13 ± 0.08 0.14 ± 0.14 0.741 0.761 0.804 Very large VLDL particles (nmol/l) 0.77 ± 0.59 0.64 ± 0.49 0.87 ± 0.84 0.58 ± 0.39 0.56 ± 0.45 0.74 ± 1.27 0.84 ± 0.60 0.83 ± 0.78 0.949 0.858 0.947 Large VLDL particles (nmol/l) 5.63 ± 3.34 5.14 ± 3.15 5.61 ± 4.36 4.28 ± 2.10 4.21 ± 2.74 5.01 ± 6.46 6.07 ± 3.82 5.88 ± 4.18 0.886 0.780 0.875 Medium VLDL particles (nmol/l) 19.7 ± 9.03 19.0 ± 8.49 18.6 ± 10.6 15.4 ± 5.10 15.6 ± 7.16 17.1 ± 15.3 20.7 ± 10.1 19.9 ± 9.63 0.938 0.724 0.794 Small VLDL particles (nmol/l) 31.9 ± 10.1 30.5 ± 9.02 30.0 ± 11.8 26.7 ± 5.87 27.9 ± 8.32 28.3 ± 13.7 32.3 ± 10.4 31.0 ± 9.30 0.875 0.739 0.761 Very small VLDL particles (nmol/l) 38.3 ± 9.63 34.0 ± 7.26 a 38.1 ± 11.3 35.8 ± 8.46 38.3 ± 7.86 36.8 ± 7.24 38.4 ± 8.34 35.2 ± 7.53 a 0.463 0.383 0.372 Mean diameter for VLDL particles (nm) 37.2 ± 1.03 37.3 ± 1.20 37.1 ± 1.58 36.9 ± 0.98 36.6 ± 1.30 36.6 ± 1.78 37.3 ± 1.52 37.5 ± 1.35 0.813 0.486 0.532 Total cholesterol in VLDL (mmol/l) 0.70 ± 0.27 0.61 ± 0.23 0.68 ± 0.30 0.59 ± 0.18 a 0.64 ± 0.17 0.64 ± 0.31 0.72 ± 0.22 0.65 ± 0.22 a 0.691 0.811 0.814 Triglycerides in VLDL (mmol/l) 1.00 ± 0.43 0.96 ± 0.39 0.96 ± 0.54 0.80 ± 0.26 0.80 ± 0.37 0.89 ± 0.79 1.04 ± 0.50 1.02 ± 0.51 0.885 0.711 0.774 IDL particles (nmol/l) 100 ± 26.1 84.3 ± 18.7 a 99.1 ± 27.5 95.9 ± 28.2 104 ± 22.0 101 ± 25.0 101 ± 27.8 92.6 ± 23.0 a 0.046 0.028 b 0.033 b Total lipids in IDL (mmol/l) 1.01 ± 0.27 0.85 ± 0.19 a 1.00 ± 0.28 0.97 ± 0.29 1.06 ± 0.23 1.02 ± 0.27 1.02 ± 0.29 0.94 ± 0.24 a 0.045 0.030 b 0.036 Large LDL particles (nmol/l) 167 ± 45.3 141 ± 31.3 a 159 ± 45.8 157 ± 48.7 1.72 ± 38.8 168 ± 44.9 167 ± 49.9 156 ± 41.8 a 0.051 0.038 0.047 Medium LDL particles (nmol/l) 138 ± 38.1 118 ± 25.3 a 126 ± 37.0 127 ± 39.8 140 ± 32.5 138 ± 37.6 138 ± 43.0 129 ± 36.6 0.103 0.109 0.132 Phospholipids in medium LDL (mmol/l) 0.19 ± 0.04 0.17 ± 0.03 a 0.18 ± 0.04 0.18 ± 0.04 0.19 ± 0.03 0.19 ± 0.04 0.19 ± 0.04 0.18 ± 0.03 a 0.047 0.035 b 0.047 Small LDL particles (nmol/l) 163 ± 41.3 141 ± 27.2 a 149 ± 40.3 151 ± 41.8 164 ± 35.6 162 ± 41.1 163 ± 46.2 155 ± 39.1 0.069 0.087 0.111 Mean diameter for LDL particles (nm) 23.5 ± 0.10 23.4 ± 0.10 a 23.6 ± 0.10 23.5 ± 0.09 a 23.5 ± 0.07 23.5 ± 0.09 23.5 ± 0.11 23.5 ± 0.12 0.555 0.449 0.431 Total cholesterol in LDL (mmol/l) 1.56 ± 0.46 1.30 ± 0.32 a 1.43 ± 0.45 1.44 ± 0.50 1.60 ± 0.42 1.55 ± 0.51 1.57 ± 0.54 1.45 ± 0.46 a 0.106 0.123 0.153 Triglycerides in LDL (mmol/l) 0.16 ± 0.04 0.15 ± 0.03 0.16 ± 0.06 0.16 ± 0.04 0.17 ± 0.03 0.17 ± 0.05 0.15 ± 0.03 0.15 ± 0.04 0.443 0.298 0.248 Serum total cholesterol (mmol/l) 4.11 ± 0.84 3.65 ± 0.65 a 4.04 ± 0.92 4.03 ± 0.92 4.23 ± 0.78 4.14 ± 0.93 4.16 ± 0.90 3.92 ± 0.76 a 0.047 c 0.036 c 0.042 Esterified cholesterol (mmol/l) 2.90 ± 0.59 2.55 ± 0.45 a 2.84 ± 0.64 2.85 ± 0.64 2.99 ± 0.57 2.91 ± 0.69 2.96 ± 0.64 2.77 ± 0.53 a 0.020 c 0.016 c 0.020 c Free cholesterol (mmol/l) 1.21 ± 0.26 1.10 ± 0.20 a 1.21 ± 0.29 1.17 ± 0.28 1.24 ± 0.23 1.23 ± 0.27 1.20 ± 0.26 1.14 ± 0.24 a 0.304 0.237 0.236 Serum total triglycerides (mmol/l) 1.39 ± 0.49 1.33 ± 0.43 1.37 ± 0.64 1.18 ± 0.30 1.20 ± 0.40 1.29 ± 0.89 1.41 ± 0.53 1.39 ± 0.57 0.765 0.777 0.800 Values are means ± SD. Differences in fold changes among the groups were tested using ANCOVA and Bonferroni’s post hoc tests. Variables in ANCOVA were log-transformed.
ANCOVA: Model 1 no adjustments, Model 2 adjusted for baseline value, Model 3 adjusted for baseline value, age, sex and use of statins. a) Change within the group, p < 0.05; b) CSO group vs. lean fish group, p < 0.05; c) CSO group vs. fatty fish group, p < 0.05. Benjamini-Hochberg false discovery rate (FDR) was used to adjust results for multiple comparisons. After FDR-adjustment p -values were no longer statistically significant. CSO, camelina sativa oil; ANCOVA, analysis of covariance.
