Clinical Nutrition 35 (2016) 907e911

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Clinical Nutrition

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Original article APOA II genotypes frequency and their interaction with saturated fatty acids consumption on profile of patients with type 2 diabetes

Neda Noorshahi a, Gity Sotoudeh b, Mahmoud Djalali a, Mohamad Reza Eshraghian c, Mohammad Keramatipour d, Marjan Ghane Basiri a, Farideh Doostan e, * Fariba Koohdani a, f, a Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran b Department of Community Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran c Department of Biostatistics and Epidemiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran d Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Iran e Department of Nutrition, Faculty of Health, Kerman University of Medical Sciences, Kerman, Iran f Diabetes Research Center, Endocrinology and Metabolism Clinical Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran article info summary

Article history: Background & aim: Several studies have suggested that APOA II-265T/C polymorphism affect lipid profile. Received 20 December 2014 The aim of this study was to investigate the effect of 265T/C APOA II polymorphism and saturated fatty Accepted 20 June 2015 acids (SFA) intake interaction on lipid profile in diabetic population who are at risk for lipid disorders. Methods: In this cross sectional study, 697 type 2 diabetic patients participated. Food consumption data Keywords: were collected using validated semi-quantitative FFQ during the last year. Realtime-PCR was used to A II determine APOA II265T/C genotypes. The interaction between the genotypes and SFA intake with lipid Diabetes type 2 fi fi pro le was tested using analysis of covariance (ANCOVA). Lipid pro les Saturated fatty acids Results: According to APOA II 265T/C (rs5082) genotype distribution results, CC genotype with a fre- quency of 12.9% and TC with that of 47.7% showed the lowest and highest frequency in our population, respectively. CC genotype subjects had significantly lower total , , Cholesterol/HDL- c ratio and non-HDL cholesterol than T allele carriers (p ¼ 0.009, p ¼ 0.02, p ¼ 0.02 and p ¼ 0.002, respectively). The interaction between genotype and SFA intake contributed to significant higher levels of LDL-c and LDL/HDL in CCs (p ¼ 0.05 and p ¼ 0.01), suggesting vulnerability of these individuals to high intake of SFA in the diet. Conclusion: APOA II polymorphism may influence the saturated fatty acid intake required to prevent dyslipidemia in the type 2 diabetic population. © 2015 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

1. Introduction the world due to its high prevalence [1]. Dyslipidemia can lead to the increased prevalence of cardiovascular disease (CVD) in these Diabetes mellitus is the most common disease caused by patients. Underlying factors in diabetes such as hyperglycemia, metabolic disorders. Today, diabetes is a major health problem in dyslipidemia and hypertension increase the risk of this outcome [2]. Studies have shown a strong association between the hyper- cholesterolemia and incidence of cardiovascular disease and its mortality so that 1 mg/dL reduction in LDL-c leads to a 1e2 percent List of abbreviation: FFQ, food frequency questionnaire; HDL-c, high density li- reduction in the relative risk of cardiovascular disease. poprotein cholesterol; LDL-c, low density cholesterol; CVD, cardiovas- cular disease; CAD, coronary artery disease; BMI, body mass index; ANOVA, analysis Nutrition is an environmental factor which has a role in initia- of variance; OR, odd ratio; ANCOVA, analysis of covariance; TRLs, triglyceride-rich tion and treatment of diabetes mellitus and its complications [3].A ; LPL, lipo lipase. healthy dietary pattern can reduce the risk of dyslipidemia and the * Corresponding author. Department of Cellular and Molecular Nutrition, School associated disorders. Saturated fatty acids consumption increases of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, the production of and the risk of atheroma. People Iran. Tel.: þ98 21 88955569; fax: þ98 21 88955698. E-mail address: [email protected] (F. Koohdani). who receive large amounts of saturated fatty acid in their diet have http://dx.doi.org/10.1016/j.clnu.2015.06.008 0261-5614/© 2015 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved. 908 N. Noorshahi et al. / Clinical Nutrition 35 (2016) 907e911 higher cholesterol levels which increase the possibility of devel- asked to report the frequency of consumption of each food item oping cardiovascular disease [4]. according to the standard size and based on option per day, week, The completed sequencing of the has identified month or year. The values for each food were converted to grams important polymorphic sites in the human genome that contribute per day using the Handbook of domestic scale [19]. Finally, the to phenotypic differences among individuals [5]. Recent genome equivalent gram of the consumption of each food item in terms of scans from different laboratories [6] have identified a locus on each person per day was measured. The N3 program was utilized to 1q21-q23 which belongs to apolipoprotein A II (APOA assess the energy and nutrient contents. Values of food items in II). Apo A II is the second most abundant protein in HDL-C, but its grams per day and nutrients were entered into IBM SPSS Statistics function is not well understood [7]. The direct effect of allele-265T/ (version 21). C on and FFA metabolism has been studied in trans- genic mice. In these studies, overexpression of human apoA II has 3. Physical activity caused a marked increase in cholesterol and/or triglyceride apoB containing lipoproteins [8e10]. However, this relationship is not Daily physical activity was evaluated in terms of metabolic detected in another study [11]. It has been shown that apoA II equivalent hours per day (MET.h.day) by a physical activity expression in the CC group was reduced in comparison with the TT questionnaire which was developed in Europe in previous studies and TC groups [12]. It has been suggested that the increased plasma and its validity was confirmed [20]. Previously, in a study con- concentration of apoA II is proatherogenic [13]. Human studies ducted by Kelishadi et al. [21] the reliability and validity of this reported no correlation [12,14] or higher [13,15] lipid profile in CC questionnaire were confirmed in adolescents in Iran. individuals. However, Birjmohun et al. showed inverse correlation between apoA II concentration and CVD incidence [15]. In contra- 3.1. Biochemical evaluation diction a caseecontrol study was demonstrated that APOA II polymorphism had a supportive preventive role against the inci- Venous blood samples were taken after 12e14 h fasting in order dence of CVD [16]. Given the controversial results of studies on the to measure serum [22]. Lipid levels were measured by effect of APOA II -265T/C polymorphism and serum apoA II levels enzymatic methods using kits (Pars Azmoon, Iran). on the lipid profile and the high incidence of lipid abnormalities in diabetic patients, this study aimed to assess the effect of APOA II- 3.2. DNA analysis 265T/C polymorphism and saturated fatty acids intake interaction on the lipid profile in Iranian diabetic population. DNA extraction was performed manually [23]. Realtime-PCR (Taqman assay) was used to determine APOA II265T/C geno- 2. Method and materials types using Applied Biosystems Step One. Due to lack of access to MGB probes, Zip Nucleic Acid (ZNA, Metabion, Germany) was uti- This study was part of a project conducted on 816 diabetic pa- lized. By this approach, the melting point of the probes increased tients. In this study, 697 type 2 diabetic patients who had the in- from 47.5and 50 Cto67C. clusion criteria were referred to diabetes centers in Tehran such as Iranian Diabetes Society, Gabric Diabetes Association, health cen- 3.3. Statistical analysis ters, Special Diseases Center in the East of Tehran, and health houses; and participated in this part of the work. After providing IBM SPSS Statistics (version 21) was used for all steps of the written and oral comments on the objective and methodology of analysis (SPSS Inc, Chicago, IL, USA). Chi-square test (X2) was used the study, written informed consent was obtained from the par- to determine the Hardy Weinberg equilibrium. First, in order to ticipants. This study was approved by the Ethics Committee of investigate the normality of quantitative variables, Kolmogor- Tehran University of Medical Sciences. oveSmirnov (one sample KS) was utilized. For the variables with p values less than 0.05, their normalized logarithms were used in all 2.1. General and anthropometric assessment tests. The normal log of cholesterol, triglycerides, LDL-c, HDL-c and BMI was used in all analyses. Chi-square was used to evaluate the Information regarding the required variables such as age, edu- difference in the percentage and Independent Student's t-test to cation, marital status, medical history (family history of diabetes, compare means. The interaction between APOA II genotypes and duration of diabetes and disease complications) and medication saturated fatty acids intake with lipid profile was tested using history (blood sugar and lipid-lowering medications or supple- analysis of covariance (ANCOVA) with controlling other con- ments) were collected using pre-tested questionnaires. Height and founding factors such as age, sex, physical activity, waist circum- weight were measured with minimum clothing and without shoes ference, body mass index, energy and fiber intake, and lipid using a Seca Falcon scale and a Seca stadiometer according to lowering drugs intake. High inter-correlated covariates (r > 0.2) standard protocols. Height and weight measurements were recor- were not used in the models. The significant level of P 0.05 was ded with an accuracy of 100 g and 0.5 cm [17], respectively. Also, considered. waist circumference was measured in the fasting condition with minimum clothing using a non-elastic tape in the middle of the 4. Results distance between the last rib and the iliac crest with an accuracy of 0.5 cm [17]. The Body mass index (BMI) was calculated as weight We evaluated 697 diabetic patients in this study. Table 1 shows (kg)/height2 (m). general characteristics and anthropometric measurements ac- cording to sex. According to our findings, genotype distribution 2.2. Dietary assessment ofAPOA II-265T/C (rs5082) in type 2 diabetes population was 39.4%, 47.7% and 12.9% for TT, TC and CC genotypes, respectively. CC and TC Usual dietary intake during the last year was measured using a genotypes had the lowest and the highest distribution. Genotype valid and reliable semi-quantitative food frequency questionnaire, frequencies were in HardyeWeinberg equilibrium. Assuming the which was included 147 food items [18]. Participants were inter- dominant effects of T allele, TT and TC genotypes were considered viewed face to face by trained nutrition experts. The subjects were as the T allele carrier groups, and were compared with homozygous N. Noorshahi et al. / Clinical Nutrition 35 (2016) 907e911 909

Table 1 Table 3 General, anthropometrical characteristic and genotype distribution of participant. Comparison of food intakes between CC genotype and T allele carrying group (TT þ TC). Variables Men Female p-value Dietary intakes/day APOA2 265T > C polymorphism p-valuea N 280 417 e TT þ TC(n ¼ 607) CC(n ¼ 90) Mean ± SDa Age (year) 54.2 ± 6.57 53.8 ± 6.56 0.4 Mean ± SD BMI (Kg/m2) 28.7 ± 4.12 29.8 ± 4.7 <0.001 Energy (Kcal) 2530 ± 760 2544 ± 901 0.9 WC (cm) 95.1 ± 9.6 90.9 ± 10.4 0.3 Total Fat (gr) 101.9 ± 4.3 101.82 ± 4.9 0.9 Physical activity 37.8 ± 6.2 37.5 ± 4.7 0.6 Protein (gr) 88.1 ± 28.2 89 ± 35.5 0.9 (Met.h.day) Carbohydrate (gr) 333.7 ± 109 338.4 ± 120.7 0.6 Energy from fat (%) 53 ± 7.4 53.6 ± 7.2 0.8 N (%)b Energy from protein (%) 14 ± 2.5 14 ± 2.5 0.7 Smoking 56 (20) 63 (15.1) 0.09 Energy from carbohydrate (%) 35.8 ± 7.8 15.4 ± 7.7 0.6 Alcohol intake 19 (6.8) 2 (0.48) <0.001 SFA (gr) 26.37 ± 9.5 25.60 ± 9.96 0.5 CAD history 100 (35.7) 157 (37.6) 0.6 MUFA (gr) 34.92 ± 16.2 33.9 ± 17.4 0.6 Overweight or obese 85 (30.3) 189 (45.6) <0.001 PUFA (gr) 25.22 ± 14 25.15 ± 15.29 0.9 APOA2 265T > C polymorphism, N (%) Total Total fiber(gr) 41 ± 18.2 44.3 ± 22.7 0.1 TT 102 (36.4) 172 (41.2) 274 (39.4) Cholesterol(gr) 220.5 ± 190.2 201.11 ± 87.45 0.3 CC 36 (12.8) 54 (13.0) 90 (12.9) DHA(gr) 0.33 ± 0.15 0.286 ± 0.17 0.7 TC 142 (50.8) 191 (45.8) 333 (47.7) EPA(gr) 0.056 ± 0.046 0.062 ± 0.055 0.1 Total 280 (100) 417 (100) 697 (100) SFA (Saturated fatty acids), MUFA (Mono unsaturated fatty acids), PUFA (Poly un- BMI (Body Mass Index), WC (Waist circumstance), CAD (Coronary Artery Disease). saturated fatty acids), DHA (Docosahexaenoic acid), EPA (Eicosapentaenoic acid). a Student's t-test. a Student's t-test. b X2 test.

non-HDL cholesterol) (Table 4). To evaluate saturated fatty acids intake we divided our population into two groups, considering CC group. Table 2 shows the general characteristics and anthro- mean saturated fatty acids intake of our population as cut point. fi pometric measurements according to APOA II-265T/C. No signi - Cholesterol and LDL/HDL levels significantly increased in CC group cant differences were observed in the general characteristics or (p ¼ 0.04 and p ¼ 0.01 respectively). However, in T carrier groups no anthropometric measurements between genotypes. Only the CC significant differences were observed between two SFA intake ¼ group had a higher mean age than T carriers (p 0.037). No sig- groups. These findings suggest vulnerability of CC individuals to fi ni cant difference was observed in food intakes of CC genotype as high intake of dietary saturated fatty acids. Obtained results from compared with T allele carrying group (Table 3). Crude comparison interaction between APOAII-265T/C genotype and saturated fatty fi ¼ of the lipid pro le showed lower total cholesterol (p 0.011), tri- acids intake on the lipid profile revealed a significant correlation ¼ ¼ glycerides (p 0.041), Chol/HDL (p 0.02) and non-HDL choles- only between LDL-c and LDL/HDL. In CC individuals who had ¼ terol (p 0.002) average in the CC than the T carrier group. The received low SFA these two parameters were lower in comparison fi results remained signi cant after adjusting for age, sex, physical with T carrier groups. When SFA intake was higher, LDL-c and LDL/ fi activity, waist circumference, energy and ber intake, BMI and HDL levels in CC individuals were higher than T carrier groups ¼ lipid-lowering medications intake (p 0.009 for total cholesterol, (p ¼ 0.05 and p ¼ 0.01 respectively) (Table 5). In the other words, CC ¼ ¼ ¼ p 0.02 for triglycerides, p 0.02 for Chol/HDL and p 0.002 for individuals who consumed high saturated fatty acids had higher LDL-C and LDL/HDL ratio than T carriers. This finding support the necessity of lower consumption of saturated fatty acids in CC in- Table 2 dividual's diet. Comparison of general and anthropometric characteristics between CC genotype and T allele carrying group (TT þ TC).

Variables APOA2 265T > C polymorphism p-value 5. Discussion TT þ TC CC fi fi N 607 90 e According to our ndings, CC genotype subjects had signi - Mean ± SDa cantly lower total cholesterol, triglyceride, Cholesterol/HDL-c ratio Age (year) 53.9 ± 6.6 55.4 ± 5.6 0.03 and non-HDL cholesterol compared to T allele carriers so their lipid BMI (kg/m2) 29.2 ± 4.5 29.2 ± 4.5 0.8 WC (cm) 92.5 ± 10.1 92.7 ± 11.9 0.8 ± ± Physical activity 37.7 5.5 37.2 4.4 0.4 Table 4 (Met.h/d) Lipid profile mean in CC and T allele carrying group (TT þ TC). N (%)b Education level 92 (15.2) 19 (21.1) Lipid profile APOA2 265T > C polymorphism p-valuea p-valueb Illiterate 389 (64.0) 53 (58.9) 0.3 CC TT þ TC Primary/high school academic or more 126 (20.8) 18 (20.0) Mean ± SD Mean ± SD Smoking 111 (18.3) 8 (8.9) 0.058 Alcohol intake 18 (2.96) 3 (3.33) 0.7 Total cholesterol(mg/dl) 175.9 ± 45.4 192.7 ± 57.4 0.011 1e0.009 Family history of diabetes 490 (80.5) 75 (83.3) 0.5 Triglyceride(mg/dl) 158.0 ± 61.3 174.6 ± 77.3 0.041 0.02 Kidney diseases history 110 (18.12) 15 (15.5) 0.6 LDL-c (mg/dl) 107.2 ± 34.7 107.8 ± 34.9 0.8 0.7 CAD history 223 (36.7) 34 (17.7) 0.7 HDL-c (mg/dl) 53.6 ± 13.3 53.0 ± 12.5 0.8 0.8 diseases history 103 (17.0) 16 (17.7) 0.8 LDL/HDL 2.06 ± 0.7 2.08 ± 0.6 0.6 0.5 Lipid lowering drugs intake 352 (58.0) 52 (57.7) 0.9 Cholesterol/HDL 3.4 ± 1.17 3.8 ± 1.37 0.02 0.02 Overweight or obese 420 (69.0) 59 (65.5) 0.2 non-HDL cholesterol 124.5 ± 50.3 145.1 ± 61.9 0.003 0.002

BMI (Body Mass Index), WC (Waist circumstance), CAD (Coronary Artery Disease). a Crude (unadjusted), Student's t-test. a Student's t-test. b Adjusted for age, sex, physical activity, waist circumference, body mass index, b X2 test. energy and fiber intake, lipid lowering drug intake, Analysis of Covariance. 910 N. Noorshahi et al. / Clinical Nutrition 35 (2016) 907e911

Table 5 genotype and fatty acids intake on profile lipid status to make the Lipid profile mean compares in SFA intake adjusted for energy in CC and TT þ TC findings more applicable. Our results showedCC individuals who groups. intake high amount of saturated fatty acids, represented higher Lipid profile SFA LDL-c and LDL/HDL levels in comparison with T carrier groups, 28.45 gr >28.45 gr p valuea p valueb p valuec however CC individuals who had consumed low SFA these two parameters were lower in comparison with T carrier groups. This Mean ± SD fi Cholesterol (mg/dl) nding support the necessity of lower consumption of saturated CC 168.1 ± 44.9 187.5 ± 45 0.04 0.1 0.1 fatty acids in CC individual's diet. TT þ TC 190.4 ± 54.9 195.1 ± 60.4 0.3 Triglyceride (mg/dl) 6. Proposed mechanism CC 156 ± 72 158.1 ± 75.8 0.9 0.8 0.8 TT þ TC 174.9 ± 73.1 174.4 ± 82.7 0.5 LDL-c (mg/dl) Animal studies have shown that high levels of apoA II inhibits CC 101.2 ± 35.8 115.3 ± 32.9 0.056 0.039 0.05 LPL; as a result, VLDL remains in plasma for hours [10,24]. The main TT þ TC 108.9 ± 3.7 106.1 ± 32 0.6 proposed underlying mechanism in most animal studies is that HDL-c (mg/dl) apoA II increases in TRLs and thus inhibits LPL activity. Similar to CC 54.3 ± 14 52 ± 12.2 0.5 0.4 0.5 fi TT þ TC 52.6 ± 11.4 53.5 ± 13.5 0.5 the nding of a survey on healthy normolipidemic women, apoA II LDL/HDL concentration was directly correlated with plasma triglyceride CC 1.93 ± 0.72 2.27 ± 0.72 0.019 0.007 0.01 concentrations and inversely related to HDL-mediated induction of þ ± ± TT TC 2.1 0.66 2.04 0.63 0.3 LPL activity. The important finding was that the fat-loading test had Cholesterol/HDL CC 3.29 ± 1.1 3.75 ± 1.1 0.06 0.09 0.1 no effect on plasma apoA-II levelsand TRLs also contained negli- TT þ TC 3.76 ± 1.3 3.8 ± 1.4 0.7 gible amounts of apoA-II. This finding strongly suggests that the non-HDL cholesterol significant increase in apoA-II in TRL which leads to LPL inhibition is CC 118.3 ± 51.6 134.7 ± 48.1 0.1 0.1 0.2 not a physiological mechanism in humans [24]. This difference can þ ± ± TT TC 142.9 60.1 147.3 62 0.3 be explained as follows: Most of the animal studies have been SFA, saturated fatty acids. conducted in either knockout or transgenic mice, which represent a In each genotype group, Student's t-test. major differences with human apoAII including variable effect on b þ Crude (unadjusted), mean compare of allele CC to T (TT TC) considering SFA fi intake, Analysis of Covariance. HDL levels, which can be attributed to species-speci c differences c Mean compare of allele CC to T (TT þ TC) considering SFA intake, adjusted for in amino acid sequences, dimer formation and affiliation for lipo- age, sex, physical activity, waist circumference, body mass index, energy and fiber protein surfaces [26]. Besides, overexpression of human apoAII in intake, and lipid lowering drug intake, Analysis of Covariance. mice has led to high concentration of apoAII in mice blood, which does not physiologically happen in human. Although our results were consistent with results from these studies and the proposed profile is more protective against atherosclerosis. The interaction mechanism of apoAII's role in TRLs lipolysis by LPL is in line with between genotype and SFA intake, shown to be attributed to higher our findings, the mentioned differences suggest that these species LDL-C and LDL/HDL in CCs, is suggestive of vulnerability of these may not be proper animal models for reflecting HDL metabolism in individuals to high dietary SFA intake. human. In this respect, Brousseau et al. have reported that trans- In the studies on human apoA II transgenic mice, it has been genic mice are not appropriate for assessing the effects of geno- revealed that overexpression of apoA II has led to an increase in the types on lipoprotein metabolism and atherosclerosis susceptibility levels of triglycerides and total cholesterol along with a significant [26]. Also, they claimed transgenic rabbit can be a unique tool for reduction in HDL-c level in comparison with non-transgenic mice this purpose [26]. Moreover most of the studies focus on apoAII [24,25]. Our results were in line with results obtained from animal effect on HDL, thereby probable role of apoAII in other lipoproteins studies and also with human studies in which reduced apoA II metabolism is incompletely understood [27]. It is probably not expression occurred in non-carriers with a CC genotype compared representative of the “quality” of the HDL particle. Recently, the to carriers with TT and TC genotypes [12]. notion of “dysfunctional HDL” has been advanced in studies of the Finding in the lipid profile are inconsistent and dialectical. HDL proteome [28]. Similar to our findings, Birjmohun [15] showed that total choles- As there is no unique finding about APOA II polymorphism effect terol, triglyceride and LDL-c were significantly lower in individuals on lipid profile in human studies, there is no described mechanism. with lower serum apoAII concentration. However, in the study What is clear is that APOA II rs5082 is found in the middle of conducted by Duesing et al. [13] it was reported that CC individuals element D, between nucleotides 255 and 276. Element D binds had marginally higher total cholesterol levels and waist-to hip ra- to several various nuclear factors that play a role in APOA II tios in the French Caucasian type 2 diabetic and normoglycemic transcription regulation [16]. The presence of the C allele is asso- individuals. This was while, Xiao et al. [16] reported no correlation ciated with nuclear binding factors in this region, resulting in between APOA II-265T/C and lipid profile. Regarding the CAD attenuation of APOAII expression in the liver cells which in turn incidence, Birjmohun findings showed inverse correlation between leads to a reduction in plasma apoAII secretion [12]. As a result, the apoAII concentration and CAD incidence [15]. In contradiction a carriers of this allele produce lower apoA II levels in response to caseecontrol study demonstrated that APOA II polymorphism had stimulatory factors such as the postprandial condition [29]. Previ- a supportive preventive role against the incidence of cardiovascular ous studies have shown that apoAII increases lecithin cholesterol disease in a white Australian population [16]. acyltransferase (LCAT) activity, boosts cholesterol efflux and lessens As in most studies food intake and its possible interaction with cholesterylester hydroperoxides; while HDL from human apoAII the genotype has not been assessed, we cannot judge that whether transgenic mice failed to guard LDL from oxidation [24]. However, these results are yielded from higher fatty acids intake, or are apoAII concentration was reported to be directly correlated with exclusively due to potential effect of these specified genotype total cholesterol, LDL-c [15] and triglyceride [13,15] and inversely groups, or both as an interaction. The effect of food intake is related to HDL-c [13,15]. Interestingly, in one study it is claimed that considered only in this study and the study performed by Corella apoAII concentration is inversely related to CAD incidence [16]. [14]. In the current study we tried to investigate the effect of both Careful considerations should be noted in interpretation of these N. Noorshahi et al. / Clinical Nutrition 35 (2016) 907e911 911 results. Due to financial problems, many factors such as plasma a novel susceptibility locus for early-onset diabetes on chromosome 3q27- concentrations of apoA II, insulin resistance, and fat tolerance test qter and independent replication of a type 2-diabetes locus on chromosome 1q21-q24. Am J Hum Genet 2000;67(6):1470e80. were not performed. In order to reach a more precise and thorough [7] Borghini I, James R, Blatter M, Pometta D. Distribution of conclusion, it is highly recommended to measure these factors in between free and A-II complexed forms in verylow- and high-density lipo- further studies. : functional implications. Biochim Biophys Acta 1991;1083(2): 139e46. In conclusion, in this study we tried to examine the interaction [8] Julve-Gil J, Ruiz-Perez E, Casaroli-Marano R, Marzal-Casacuberta A, Escola- of saturated fatty acid intake and APOA II polymorphism on the Gil J, Gonzalez-Sastre F, et al. Free cholesterol deposition in the cornea of lipid profile in type 2 diabetic patients. The results showed that human apolipoprotein A-II transgenic mice with functional lecithin:choles- terol acyltransferase deficiency. Metabolism 1999;48(4):415e21. LDL-c and LDL/HDL levels increase in response to saturated fatty [9] Escola-Gil J, Julve J, Marzal-Casacuberta A, Ordonez-Llanos~ J, Gonzalez- acids intake in CC group. Besides, in high consumption of saturated Sastre F, Blanco-Vaca F. ApoA-II expression in CETP transgenic mice increases fatty acids, these parameters were also higher in the CC group when VLDL production and impairs VLDL clearance. J Lipid Res 2001;42(2):241e8. þ [10] Boisfer E, Lambert G, Atger V, Tran N, Pastier D, Benetollo C, et al. Over- compared to the T carrier groups (TT TC). In other words, when expression of human apolipoprotein A-II induces hypertriglyceridemia due to saturated fat intake is high (>28.45 g/day), significant differences in defective very low density lipoprotein hydrolysis. J Biol Chem 1999;274(17): LDL-c and LDL/HDL levels were detected between CC individuals 11564e72. and T-allele carriers. It suggests that non-carriers may be vulner- [11] Tailleux A, Bouly M, Luc G, Castro G, Caillaud J, Hennuyer N, et al. Decreased susceptibility to diet-induced atherosclerosis in human apolipoprotein A-II able to high saturated fatty acids in their diet. Altogether, the results transgenic mice. Arterioscler Thromb Vasc Biol 2000;20(11):2453e8. can be used in detecting high risk genotypes, designing screening [12] van't Hooft F, Ruotolo G, Boquist S, Faire U, Eggertsen G, Hamsten A. Human programs, providing nutritional advice and plans for prevention evidence that the apolipoprotein A-II gene is implicated in visceral fat accu- mulation and metabolism of triglyceride-rich lipoproteins. Circulation and treatment of lipid disorders and their associated complications 2001;104:1223e8. in these patients. [13] Duesing K, Charpentier G, Marre M, Tichet J, Hercberg S, Balkau B, et al. Evaluating the association of common APOA2 variants with type 2diabetes. BMC Med Genet 2009;10(13). Author contributions [14] Corella D, Arnett D, Tsai M, Kabagambe E, Peacock J, Hixson J, et al. The _256T_ C polymorphism in the apolipoprotein A-II gene promoter is associated with FK conceived and designed the study, NN, FK and MD partici- body mass Index and food intake in the genetics of lipid lowering drugs and diet network study. Clin Chem 2007;53(6):1144e52. pated in its performance, MRE, NN and FK performed the statistical [15] Birjmohun R, Dallinga-Thie G, Kuivenhoven J, Stroes ES, Otvos J, Wareham N, analysis. NN, GS, FK and MGB drafted and edited the manuscript. et al. Apolipoprotein A-II is inversely associated with risk of future coronary NN, GS and FD conducted the literature search, participated in its artery disease. Circulation 2007;116:2029e35. [16] Xiao J, Zhang F, Wiltshire S, Hung J, Jennens M, Beilby J, et al. The apolipo- design and coordination, FK, MK and MD provided critical input protein AII rs5082 variant is associated with reduced risk of coronary artery during manuscript preparations. NN and FK performed data inter- disease in an Australian male population. Atherosclerosis 2009;199(2):333e9. pretation and wrote the manuscript. All authors read and approved [17] WHO. Waist circumference and waist-hip ratio: report of a WHO expert the final manuscript. consultation. Geneva. December 2008. 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