Clinical Nutrition 35 (2016) 907e911 Contents lists available at ScienceDirect Clinical Nutrition journal homepage: http://www.elsevier.com/locate/clnu Original article APOA II genotypes frequency and their interaction with saturated fatty acids consumption on lipid 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 À265T/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 Apolipoprotein A II determine APOA IIÀ265T/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 À 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, triglyceride, 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 lipoprotein 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 lipoproteins; LPL, lipo protein 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 chylomicrons 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 human genome 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 chromosome 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 triglycerides 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 lipids [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 IIÀ265T/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.