Accepted Manuscript

Dietary n-3 Polyunsaturated Fatty Acid intake modulates impact of Insertion/Deletion

polymorphism of ApoB gene on obesity risk in type 2 diabetic patients

Masoumeh Rafiee, MS.c, Gity Sotoudeh, Ph.D, Mahmoud Djalali, Ph.D, Ehsan
Alvandi, MS.c, Mohammadreza Eshraghian, Ph.D, Fereshteh Sojoudi, BS.c, Fariba
Koohdani, Ph.D
PII: S0899-9007(16)30004-1
DOI: 10.1016/j.nut.2016.03.012
Reference: NUT 9746

To appear in: Nutrition

Received Date: 10 December 2015

Revised Date: 21 February 2016
Accepted Date: 15 March 2016

Please cite this article as: Rafiee M, Sotoudeh G, Djalali M, Alvandi E, Eshraghian M, Sojoudi F,
Koohdani F, Dietary n-3 Polyunsaturated Fatty Acid intake modulates impact of Insertion/Deletion
polymorphism of ApoB gene on obesity risk in type 2 diabetic patients, Nutrition (2016), doi: 10.1016/
j.nut.2016.03.012.

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 ACCEPTED MANUSCRIPT

Dietary n-3 Polyunsaturated Fatty Acid intake modulates impact of

Insertion/Deletion polymorphism of ApoB gene on obesity risk in type 2
diabetic patients
Running title: n-3 PUFA modulates impact ApoB Ins/Del on obesity

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Masoumeh Rafiee MS.c 1 , Gity Sotoudeh Ph.D 2 , Mahmoud Djalali Ph.D 1, Ehsan Alvandi MS.c 1 ,

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Mohammadreza Eshraghian Ph.D 3 , Fereshteh Sojoudi BS.c 4, Fariba Koohdani Ph.D1*

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1. Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical
Sciences, Tehran, Iran.
2. Department of Community Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences,

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Tehran, Iran.
3. Department of Biostatistics and Epidemiology, School of Public Health, Tehran University of Medical Sciences, Tehran,
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Iran.
4. School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran.
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Word count: 4492

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Number of tables: 4
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*Corresponding author: Fariba Koohdani, Department of Cellular and Molecular Nutrition, School of
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Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran. Phone Number:

+98-21-88955569(105) - 09122182802, Fax: +98-21-88955698, email: fkoohdan@sina.tums.ac.ir

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Abstract
Objectives: To determine whether dietary n-3 PUFA intake modulates the association between ApoB Ins/Del
polymorphism and obesity in type 2 diabetic patients.

Methods: In this cross-sectional study, 700 patients with type 2 diabetes were recruited in Tehran. Weight and WC

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(waist circumference) were measured and BMI (body mass index) calculated. Dietary intake was assessed by using a
validated semi-quantitative food frequency questionnaire. ApoB genotyping was performed by 8% polyacrylamide gel
electrophoresis (PAGE).

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Results: We observed a significant Ins/Del genotype-dietary n-3 PUFA interaction for BMI, WC and obesity risk in both
unadjusted (P= 0.007, P=0.001, P= 0.021, respectively) and adjusted (P= 0.007, P=0.04, P= 0.002, respectively). Thus,

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the carriers of the Del allele were only associated with lower BMI (p=0.01) and WC (P=0.002) among individuals with
high n-3 PUFA intakes (≥ 0.6% of energy), but not in group with low n-3 PUFA intake (< 0.6%). Also, when dietary n-3
PUFA was < 0.6%, general obesity risk in carriers of the Del allele was about 1.6 times higher than Ins/Ins homozygotes

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(OR=1.59; 95% CI 1.05–2.52, P = 0.039). But in high n-3 PUFA intake (≥ 0.6%) was 0.46 times lower (OR=0.46; 95% CI
0.25–0.79, P = 0.003). Moreover, similar interaction was found in central obesity only in men after adjustment for
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confounder variables (P=0.041).

Conclusion: These findings support that a diet with high n-3 PUFA (≥ 0.6%) can decline of the obesity risk in carriers of
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the Del allele of ApoB gene.

Keywords: ApoB Ins/Del, Obesity, n-3 PUFA, diabetes.

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Highlights:
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►We identified an interaction between ApoB Ins/Del genotypes and n-3 PUFA intake.
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►Participants carrying Del (Ins/Del, Del/Del) may benefit of diet with high n-3 PUFA.

►The genetic variants had no significant influence on the change in obesity-variables.

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►The present results may contribute to personalized dietary interventions.

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Abbreviations:
ApoB, apolipoprotein B; FFQ, food frequency questionnaire; PCR, polymerase chain reaction; BMI, body mass
index; ANOVA, analysis of variance; OR, odd ratio; ANCOVA, analysis of covariance.
Funding:
This research was funded and supported by the Tehran University of Medical Sciences (Research project number

91-03-161-19322).

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Introduction
Prevalence of obesity has increased remarkably in the world wide [1]. Obesity has been associated with chronic disease
such as type 2 diabetes and also its related complications, which including dyslipidemia, hypertension and
cardiovascular disease (CVD) [2].In this regard, reported that more than 85% of diabetic patients are obese [3]. Obesity
can be resulted as interaction between environmental and genetic factors; estimated percent intervals for genetic

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factors are 40-75 [4].Nutrition is one of the most important environmental factors that interacts with genes to increase
or decrease of obesity risk [5]. Overall, Nutrigenetics refers to the differing phenotypic impacts of varied diet on
subjects with the same genotype [6]. N-3 polyunsaturated fatty acid (PUFA), one of the dietary factors that has great

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importance in modifying gene expression involved in lipid metabolism and obesity [7].Recently, studies that have
evaluated a gene–diet interaction of n-3 PUFA as a main dietary component is used [8,9]. An investigation in world

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demonstrated that dietary n-3 PUFA intake to have prevention impact on the initiation and advancement of obesity
[10]. Human genome scan project have been identified that there are several genes associated with human obesity
[11]. In fact, genes could have effect on obesity in affecting energy homeostasis, dietary nutrient metabolism and

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hypothalamic appetite signaling pathway [12]. ApoB is one of the important genes that characterized several single
nucleotide polymorphisms (SNPs) related to type 2 diabetes, obesity, dyslipidemia and CVD risk [13]. ApoB is the major
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protein for the synthesis and secretion of both chylomicrons and VLDL, and acts as a ligand for LDL receptor to facilitate
the uptake of serum cholesterol [14]. Therefore, there is well reasoned to investigate the association between obesity
and genetic variants of lipid regulatory protein, since obesity is excessive fat accumulation in the body tissues and cells
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[15]. ApoB gene is located on chromosome 2P, and contains 29 exons. The Insertion/Deletion polymorphism is within
the first exon. The frequent allele (93bp) encodes a 27 amino acid peptide and the deletion of 9bp results in 24 amino
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acid peptide [16]. Inadequate data are available on impacts of this polymorphism of ApoB gene related to obesity and
the responsiveness to dietary intakes. Saha et al. in Hindi healthy population in Singapore reported that Del allele
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carriers (Ins/Del + Del/Del) compare with Ins/Ins genotype have higher BMI (Body mass index) [17]. On the other hand,
Yatsyshina et al. no observed a significant association between ApoB Ins/Del polymorphism and obesity in Russian
population [18]. With considering, conflicting observation have been reported on the association between the ApoB
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Ins/Del polymorphism and obesity, suggesting that environmental factors such as dietary factors may be affected on
this association. The few studies conducted in this field. In only interaction studies observe an impact of the ApoB
Ins/Del polymorphisms on plasma lipid responses to dietary change [19]. To the our knowledge, there are no study and
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data that show interaction between ApoB Ins/Del polymorphism and dietary intake on obesity risk. Thus, in the current
study, It has been determined distribution of ApoB Ins/Del polymorphism and in the same line, the critical question
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that“ Is there any possibility to be modified an association between ApoB Ins/Del polymorphism and obesity risk by
dietary intakes especially n-3 PUFA in type 2 diabetic patients or not”

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Materials and Methods

Study Design and Subjects

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This study has been previously described [20].The study sample consisted of 700 patients with type 2 diabetes (276
men and 424 women), aged 35–65 years, who were recruited randomly from the Iranian Diabetes Society, Gabric

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Diabetes Association and other Health Centers in Tehran city. Diagnosis of patients with type 2 diabetes were based on
fasting blood glucose ≥126 or consumption of drugs for diabetes treatment or both. Criteria for exclusion included age

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<35 and over 65 years, insulin use, alcohol consumption 24 hours before blood sample collection, pregnancy and
lactation. The study was approved by the Ethics Committee of Tehran University of Medical Sciences, and all patients
provided written informed consent for participation in this study.

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Anthropometric and physical activity determinations
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Anthropometric data were measured by standard methods. Weight and height were measured with minimal clothing
without shoes using Seca falcon scales (Seca, Germany) with an accuracy of 100g and height gauges with an accuracy of
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0.5 cm, respectively. Waist circumference was measured by using non-elastic tape with an accuracy of 0.5 cm in the
midpoint of between the upper edge of the iliac crest and the lower edge of the chest and the last rib [21].
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Measurements were determined by a dietitian. BMI was calculated by dividing weight (in kg) to the square of height (in
m) scale. We according to WHO classifications defining cases of general and central obesity as those individuals having
BMI ≥ 30 kg/m2 , WC ≥90 cm for men and ≥80 cm for women, respectively [22]. Physical activity was measured by
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questionnaire and metabolic equivalent hour per day has been calculated [23]. This questionnaire was developed and
validated in previous studies in Europe [24]. In Iran, reliability and validity of the questionnaire have been confirmed by
the Kelishadi et al. [25].
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Dietary and general information

Dietary intakes were assessed using a semi-quantitative food frequency questionnaire. The questionnaire was designed
in Tehran Lipid and Glucose Study and includes 148 food items. Reliability and validity of the questionnaire have been
confirmed in Iran [26]. This questionnaire designed base on frequency of country usual foods that consumed during the
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past years (times in daily, weekly, monthly and annual). Food items were defined according to the US Department of
Agriculture (USDA) standard portion sizes. Means daily intake of each item was calculated by multiplying the
consumption frequency of each food by its standard portion size from the list exchange [27]. Eventually, the grams of
each food item were calculated. Also, this dietary intake was adjusted for total energy intake by the nutrient density
procedure (percent of energy from dietary carbohydrate, protein, total fat, SFA, MUFA, total PUFA, n-3 PUFA and n-6
PUFA). Intakes were categorized into two groups according to the median value, below and above or equal. N3

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software was used to calculate the energy and nutrient intake that designed for Iranian foods. Three categories of
smoking status were used: current (including irregular smoking), former and never in last 6 month. Alcohol habits were

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divided into two categories including Individuals no and yes alcohol consumption during the last 6 month in the
questionnaire.

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Genetic analysis

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Blood samples were collected for the determination characterization of genomic DNA [28]. Genotyping for ApoB Ins/Del
was conducted by using a Polymerase Chain Reaction (PCR) in in 25 μL mixture, containing 50 ng of DNA, 0.2 mM of
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each primer Forward (5’-CAGCTGGCGATGGACCCGCCGA-3’) and Reverse (5’-ACCGGCCCTGGCGCCCGCCAGCA-3’), 2X Taq
polymerase mix (SinaClon Bioscience Co.), and 7% Dimethyl sulfoxide (Sigma-Aldrich Co.) PCR condition was 94°C for 50
second and 64°C for 90 seconds in 35 cycles. Genotypes determined by 8% polyacrylamide gel electrophoresis [29].
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Statistical analysis
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Statistical analysis was determined by SPSS software version 18 (IBM SPSS Inc., Chicago, IL, U.S.). Chi-square (x2) test
was applied to characterize the Hardy Weinberg equilibrium. Data normality examined by Kolmogorov–Smirnov Test.
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Variables that have not normal distribution expressed to log-transformed. According, we analyzed variables including
lifestyle and dietary intake among two group genotypes (Ins/Ins and Ins/Del + Del/Del) using Chi-square test and t-test.
The association between the ApoB Ins/Del polymorphism and Anthropometric determinations was analyzed by
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independent t-test in crud model and ANCOVA (analysis of covariance) in adjusted model for age, gender, family
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relationship, smoking , physical activity and daily energy intake. The interaction between dietary intakes (Carbohydrate,
protein, SFA, MUFA, total PUFA, n-3 PUFA, n-6 PUFA and cholestrol according median value) and ApoB Ins/Del
polymorphism on BMI and WC measurments were evaluated by using ANCOVA multivariate, adjustment for
confounding variables including age, gender, BMI, family relationship, physical activity, total energy intake and
percentage of macronutrients and fiber intake. In addition, the dietary n-3 PUFA-genotype interaction on obesity risk
has been tested by logistic regression adjusting for age, gender, family relationships,smoking, physical activity, daily
energy intake, and percentage of carbohydrate and protein . These analyses for general obesity risk were determined in

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whole of the population, but for central obesity risk in men and women separately in order to different the cutoff
points. Values of P<0.05 were expressed statistically significant.

Results
The allelic and genotypic frequency is shown in Table 1 based on gender. The genotype distributions were in Hardy-

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Weinberg equilibrium (X2= 0.046, P=0.97). We did not find significant associations between this polymorphism and
General factors and dietary intake (Table 2). Also, no significant association was found between the ApoB Ins/Del
polymorphism and measurement related-obesity including Weight , Height, BMI, Waist circumference in both crude

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and multivariate adjusted model (adjusted for age, gender, family relationship, smoking, physical activity and daily
energy intake; P>0.05) (Table 3). Therefore, we have tested the interaction between ApoB Ins/Del polymorphism and
dietary intakes in modulating obesity. The carbohydrate, protein, total fat, saturated fatty acid, MUFA, PUFA, n-3 PUFA,

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n-6 PUFA and cholesterol were categorized into two groups, according to their medians of the population (54, 14, 35, 9,
12, 8, 0.6, 7 percent and 196 mg, respectively). We found a gene-diet interaction between the ApoB Ins/Del and n-3
PUFA intake in relation to the BMI (P=0.007) and WC (P=0.001) that remained statistically significant after the

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adjustment of this multivariate model for age, gender, BMI, family relationship, physical activity, total energy intake
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and protein, carbohydrate and fiber intake (P= 0.007 and P=0.04, respectively) (Table 4). The Del allele carriers had
significantly lower BMI (P=0.01) and WC (P=0.002) only in the group of high dietary n-3 PUFA (≥ 0.6 of energy), whereas
had a non-significant light increase in BMI and WC in group of low dietary n-3 PUFA intake (< 0.6 of energy). No gene-
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diet interaction on BMI and WC has been shown with others dietary intakes under our study. Eventually, the impact of
the ApoB Ins/Del -n-3 PUFA interaction on the general and central obesity risk was tested (Table 5). When was
analyzed, we identified a statistically significant interaction between the n-3 PUFA intake and ApoB Ins/Del on the
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general obesity in the crude model (P=0.021) and also after the adjustment for age, gender, family
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relationships,smoking, physical activity, daily energy intake, and percentage of carbohydrate and protein (P= 0.002).
When the dietary n-3 PUFA intake was low (< 0.6% of energy), subjects with the Del allele had about over 1.5 fold the
risk of obesity (OR=1.59; 95% CI 1.05–2.52, P = 0.039) compared with the Ins/Ins homozygotes. Whereas, in high the n-3
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PUFA intake (≥ 0.6 of energy), carriers of the Del allele had about half risk of obesity (OR=0.46; 95% CI 0.25–0.79, P =
0.002) compared with the Ins/Ins subjects. Also, we found a significant interaction between the ApoB Ins/Del
polymorphism and n-3 PUFA intake on the risk of central obesity in males (P= 0.041) in a multivariate adjusted model.
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At low n-3 PUFA intake (< 0.6% of energy), carriers of the Del allele had a significant higher risk of central obesity
(OR=2.5; 95% CI 1.05–6.02, P = 0.039). While at high n-3 PUFA intake (≥ 0.6 of energy), these carriers had a non-
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significant lower risk of central obesity in males (OR=0.48; 95% CI 0.16–1.3, P = 0.12) compared with Ins/Ins
homozygotes. There is no significant interaction on central obesity risk for females was observed in both models
adjusted and unadjusted (P=0.16 and P=0.07, respectively).

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Discussion
In the present study, we observe a novel interaction between ApoB Ins/Del polymorphism and n-3 PUFA intake on the
BMI, and WC. When n-3 PUFA intake was ≥ 0.6% (intake median) of total energy, in the present study, we observe a
novel interaction between ApoB Ins/Del polymorphism and n-3 PUFA intake on the BMI, WC and obesity risk. When n-3
PUFA intake was ≥ 0.6% (intake median) of total energy, subjects with Ins/Del and Del/Del genotypes displayed

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significantly lower BMI and general obesity risk in all populations and non-significantly lower central obesity risk only
in men compare with Ins/Ins homozygous. On the other hand, in low n-3 PUFA intake (< 0.6% of total energy), carriers
of Del allele was not associated with BMI, WC but had 1.59 and 2.5 fold higher risk for general and central obesity than

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Ins/Ins genotype. To our knowledge, there is no study that shows interaction between ApoB Ins/Del and diet on obesity
risk. In the only investigation close to our study, Jemaa et al. examined ApoB Ins/Del -diet interaction on lipid profile.

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The results of their study exhibited that carriers Del allele especially the homozygote had the greatest LDL-c
concentration in initial and also significant and great reduction in response to a low-calorie diet compare with subjects
Ins/Ins genotype [30]. Except n-3 PUFA intake in our study, no interactions identified for carbohydrate, protein, total

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fat, SFA, MUFA, total PUFA, n-6 PUFA and cholesterol of diet on anthropometric measurement (BMI and WC) and risk of
obesity. However, this significant interaction was a dose-dependent impact of dietary n-3 PUFA intake. In fact, we
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observed that consumption of dietary n-3 PUFA in reducing BMI, WC and obesity risk in different genotypes of ApoB
Ins/Del polymorphism both unadjusted and adjusted models was dose-dependent. Nutritional genomics contribute to
access efficient and suitable dietary for prevention and treatment of disease such as obesity and detection of subjects
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who will advantage from personalized dietary recommendations (PDRs) [31]. Interesting, the obesity risk decreased in
carriers of Del allele with consumption of high n-3 PUFA in diet. The molecular mechanism expressing the association of
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Ins/Del polymorphism of ApoB gene and obesity and interaction those with diet is not explored clearly yet. The loss of
three amino acids in the Del allele can affect the hydrophobicity of the signal peptide [32]. Hence, experimental
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investigation has been demonstrated that carriers Del allele of ApoB gene has defective assembly and production in
very low density lipoprotein (VLDL), which result in storage of fat in liver tissue and down-regulation of secretion of LDL-
receptor [33]. This reduction in LDL-receptor in carriers Del allele of ApoB gene result in enhanced hepatic triglyceride
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and ApoB -100 production and decrease clearance them [34]. Therefore, increasing TG-enriched lipoprotein and TG in
liver in subject with Del allele result in more uptake of TG by adipose tissue [35]. Investigations demonstrated that
serum TG concentration was a positive association with hypertrophy and hyperplasia of adipocytes [36]. Also, observed
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that loss synthesis of ApoB in body may increase production of leptin, which result in resistance and finally obesity [37].
This is suggesting and probably mechanism for increase obesity risk in carriers Del allele. Interestingly, animal studies
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and epidemiological observation in humans indicate the anti-obesity impacts of n-3 PUFA [38, 39, 40, and 41]. The
possibility mechanism n-3 PUFA in reduction of obesity, which including 1. Enhancing thermogenesis and lipid
catabolism through by up-regulation of genes associated with B-oxidation including carnitine palmitoyl transferase I
(CPT-1), 5`-AMP-activated protein kinase (AMPK), peroxisomal acyl-CoA oxidase (PACO), uncoupling protein 2 (UCP-2) in
liver, skeletal muscle, intestinal and adipose tissues 2. Decreasing appetite 3. Reduction synthesis and storage of fat
with down-regulation of FAS (fatty acid synthesis) [42, 43, 44 and 45]. The researchers in their current findings provide
evidence supporting that the decrement in obesity risk and obesity-related traits were associated with reduction of
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plasma TG levels that were by consumption of medium and high n-3 PUFA in diet [46]. Our findings revealed that high
n-3 PUFA intake in carriers Del allele result in reducing central obesity risk only in men.This difference between the
observed association can be explained as: several animal studies have been elucidated that hypertriglyceridemia and
central obesity related with its due to deficiency LDL-receptor in carriers of Del allele in male to be more than female
mice [35, 47]. Additionally, researchers found that the risk of central obesity is different in both genders, with more
prevalent in men [48]. This difference could be attributed to differences in sex hormones and fat distribution in both

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genders. One of the strongest reasons for the increased risk of central obesity in men, lack of estrogen hormone in
them [49, 50, 51]. Finally, the high tendency of men compare with women, independent and booster effects of

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genotypes with high risk (Ins/Del and Del/Del) and low intake of n-3 PUFA could be supported of significant increase risk
of central obesity in this group. According to the results of our study, can be explained that high intake of dietary n-3
PUFA through the above mechanisms may be result in diminished the increased risk of obesity in carriers of Del allele.

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There were some main limitations in our study, which should be characterized. Using of food frequency questionnaire
(FFQ) for dietary analysis may provide to recall bias. Our cross-sectional study cannot express credible document for
causality of the observed interaction. Also, due to time limits and also lack of enough budgets, we were unable a

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replication of the results in independent materials. Nonetheless, we did not find interactions between ApoB Ins/Del and
dietary intake including carbohydrate, protein, total fat, SFA, MUFA, total PUFA, n-6 PUFA and cholesterol on BMI and
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WC. Post- power calculations showed that these finding may be affected by insufficient sample size. Authors suggest
further studies with larger sample, since relevant interaction calculation generally requires more bigger population
samples.
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Conclusion:
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In conclusion, this study indicated an interaction between ApoB Ins/Del genotypes and n-3 PUFA intake on
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measurements of BMI and WC and obesity risk in T2DM patients. Personalized dietary recommendations (PDRs) could
be developed by result of our study. In the future Nutritionists may be able to achieve effective and quickly dietary
information base on an each person's genetic profile. Our investigation supports the high dietary n-3 PUFA intake can
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disregard the raising effect of Del allele ApoB gene on obesity and traits related-its (BMI and WC) in group low n-3 PUFA
intake, which may protect against the development of type 2 diabetes and complications related with obesity. The
amount of dietary n-3 PUFA (>median) in population of our study that indicated interaction effects was exactly based
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on minimum range of Recommended Dietary Allowances, which is equivalent to the consumption of 1.3 grams per day
or 0.6 % of total energy.To our knowledge, this study is the first investigation that determined the impact of diet and
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ApoB Ins/Del genotypes interaction on obesity risk. Suggest the dietary n-3 PUFA is a principal environmental factors in
gene-diet interaction on obesity risk.

Acknowledgments

This research was funded and supported by the Tehran University of Medical Sciences (Research project number 91-03-
161-19322). The authors wish to thank the participants in this study.

Conflict of Interest: None declared.

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Table 1. Distributions of genotype and allele frequencies of the Ins/Del polymorphism of the Apo-B gene

in diabetic men and women

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Genotype Relative allele frequencies (%) Genotype frequencies N (%)

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Del Ins Del/Del Ins/Del Ins/Ins

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Male 16.7 83.3 7(2.5) 78(28.3) 191(69.2)

Female 19 81 17(4.0) 129(30.4) 278(65.6)

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Total 18 82 24(3.4) 207(29.5) 470(67.1)
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2
X =0.046
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p =0.9
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*Statistically significant association between genotype and allele frequencies (Chi-square test)
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Table 2. General and dietary intake according to ApoB Ins/Del genotype

ApoB Ins/Del

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Ins/Ins (n=470) Ins/Del + Del/Del (n=230) P value

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Age, years 54.1 ± 6.6 53.8 ±6.5 0.57

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Physical activity, MET h/day 38 ±5.66 37.3 ±4.97 0.11

Family history of diabetes, n (%) 382 (81.4) 187 (81.3) 0.96

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Lipid lowering drugs intake, n (%) 253 (54.1) 134 (58.3) 0.29
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Glucose lowering drugs intake, n (%) 191 (40.7) 84 (36.5) 0.28

Alcohol consumption, n (%) 20 (4.3) 2 (1) 0.01

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Smoking, n (%) 88 (18.8) 46 (20) 0.69

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Total energy intake, (kcal/day)

2605 ±944 2584 ±1020 0.78
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Carbohydrate, (g/day)
349.7 ±54.6 358.58 ±62.04 0.20

Protein, (g/day)
82.5 ±16.7 83.81 ±17.7 0.36
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Total fat, (g/day)

107.67 ±25.22 104.8 ±27.9 0.17

Saturated fattyy acids, (g/day)

29.8 ±6.87 30 ±7.5 0.72
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Monounsaturated fatty acids, (g/day)

26.95 ±11.08 26.57 ±11.9 0.68

Polyunsaturated fatty acids, (g/day)

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25.21 ±10.15 24.3 ±9.96 0.27

N-3 Polyunsaturated fatty acids, (g/day)

1.6 ±0.9 1.5 ±1.04 0.55

N-6 Polyunsaturated fatty acids, (g/day)

22.4 ±13.6 21.3 ±12.23 0.31

Cholestrole, (mg/day)
225.4 ±213.4 220.4 ±109.26 0.73

Dietary fiber, (g/day)

42.5 ±25.23 42.7 ±22.08 0.91

Data are mean ±SD or n (%)

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Table 3. Anthropometric variables according to ApoB Ins/Del genotype

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ApoB Ins/Del

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Ins/Ins (n=470) Ins/Del + Del/Del (n=230) P value P value

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Weight, kg 77.06 ± 14.34 75.56 ± 13.53 0.18 0.15

Height, cm 161.53 ± 9.15 160.75 ± 9.24 0.29 0.24

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BMI, kg/m 28.9 ±4.84 29.19 ±4.31 0.40 0.10
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Waist circumference, cm 92.73 ±10.96 91.58 ±10.14 0.20 0.15
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Data are mean ±SD or n (%)

*
P value unadjuste
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P value after adjusted for age, gender, family relationship, smoking , physical activity and daily
energy intake.
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Table 4. BMI and WC stratified by ApoB Ins/Del genotype and categories of energy from diatary intakes
BMI WC

I/I (n=470) I/D+D/D (n=230) P* P**for I/I (n=470) I/D+D/D P* P**for

interaction (n=230) interaction
a b a b
Carbohydrate,% 0.86 0.92 0.60 0.32

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<54 29.9 ±4.97 29.7 ±4.46 0.70 93.4 ±10.87 91.83 ±10.83 0.20

≥54 29 ±4.67 28.7 ±4.14 0.51 92 ±11.05 91.3 ±9.5 0.58

a b a b

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Protein,% 0.64 0.57 0.96 0.81

<14 29.56 ±5 29.47 ±4.45 0.85 92.76 ±11.61 91.48 ±9.87 0.35

≥14 29.43 ±4.1 29.08 ±3.82 0.37 92.85 ±10.38 91.72 ±10.03 0.33

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a b a b
Total fat,% 0.65 0.71 0.97 0.50

<35 29.22 ±4.6 28.61 ±3.92 0.23 92.30 ±10.83 91.11 ±8.78 0.25

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≥35 29.78 ±5.01 29.65 ±4.41 0.81 93.17 ±11.13 92 ±11.05 0.35
a b a b
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SFA,% 0.51 0.60 0.98 0.53

<9 29.37 ±4.6 28.6 ±4.02 0.21 92.45 ±9.92 90.84 ±10.30 0.32

≥9 29.71 ±5.03 29.61 ±4.5 0.84 93.13 ±11.82 91.95 ±10.15 0.33
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a b a b
MUFA,% 0.73 0.81 0.92 0.76

<12 29.16 ±4.6 29.64 ±4.11 0.44 92.14 ±10.47 90.93 ±9.78 0.33
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≥12 29.88 ±5.08 29.75 ±4.42 0.82 93.43 ±11.51 92.4 ±10.75 0.45
a b a b
PUFA,% 0.43 0.42 0.07 0.14
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<8 29.28 ±4.68 29.25 ±4.64 0.99 91.77 ±10.55 92.19 ±10.50 0.71

≥8 29.70 ±5 29.11 ±3.92 0.27 93.53 ±11.3 90.83 ±9.72 0.03

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a b a b
n-3 PUFA,% 0.007 0.007 0.001 0.04

<0.6 29.12 ±4.5 29.82 ±4.48 0.20 91.87 ±10.8 93.43 ±9.19 0.22

≥0.6 29.91 ±5.22 28.51 ±4.05 0.01 93.61 ±11.12 89.45 ±10.9 0.002
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a b a b
n-6 PUFA,% 0.55 0.49 0.07 0.13
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<7 29.4 ±4.8 29.31 ±4.77 0.83 91.84 ±10.61 92.32 ±10.44 0.77

≥7 29.62 ±4.94 29.08 ±3.82 0.32 93.6 ±11.34 90.86 ±9.9 0.03
a b a b
Cholesterol, mg 0.94 0.10 0.11 0.06

<196 29.10 ±4.74 28.75 ±3.95 0.49 92.13 ±11.15 89.56 ±9.54 0.03

≥196 29.91 ±4.91 29.61 ±4.62 0.58 93.38 ±10.76 93.51 ±10.41 0.91
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All dietary data were categorized by their median value of population. Data are adjusted geometric mean (95% confidence
interval).

MUFA, monounsaturated fatty acid; PUFA, polyunsaturated fatty acid; SFA, saturated fatty acid.
*
P for genotype
**
P for interaction that obtained from multivariate model 1(a) unadjusted and model 2 (b) after adjustment for age, gender,
BMI, family relationship, physical activity, total energy intake and percentage of macronutrients and fiber intake.

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Table 5. Odds ratios (OR) and 95% confidence interval (95% CI) for the central and general
obesity stratified by ApoB Ins/Del polymorphism and the median value of n-3 PUFA intake

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n-3 PUFA intake < 0.6% n-3 PUFA intake ≥ 0.6%

P** for interaction

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* *
OR 95% CI P OR 95% CI P Model 1 Model 2

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General obesity

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I/I 1 1
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I/D+D/D 1.59 (1.05-2.52) 0.039 0.46 (0.25-0.79) 0.002 0.021 0.002
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Central obesity (Males)

I/I 1 1
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I/D+D/D 2.50 (1.05-6.02) 0.039 0.482 (0.16-1.3) 0.125 0.074 0.041

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Central obesity (Females)

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I/I 1 1

I/D+D/D 1.11 (0.57-2.22) 0.822 0.569 (0.35-1.36) 0.112 0.166 0.071

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PUFA, polyunsaturated fatty acid.

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*
P value for the genotype
**
P for interaction that obtained in the corresponding logistic regression model 1 unadjusted and model 2 after adjusting for
age, gender, family relationships,smoking, alchol consumption, physical activity, daily energy intake, and percentage of
carbohydrate and protein .