Effect of Intermitting Fasting on Lipid Profile and Hematological Parameters in Healthy Volunteers in Jordan

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1 Effect of Intermitting Fasting on Lipid Profile and Hematological Parameters in Healthy Volunteers in Jordan Mohammad A. Fararjeh, Abdulrahim AlJamal, "Mo'ez Al-Islam" E. Faris, Ref'at A. Al-Kurd, Mohammad Khalil and Yasser Al-Bustanji Abstract The Purpose of this study is to show the effects of fasting on lipid profile and hematological parameters in healthy volunteers. Eighty Healthy subjects (50 male and 30 female) volunteered to participate in all of three stages of the study. Volunteers aged from 18 to 60 were included. Blood samples were collected as follows, each volunteer at each visit gives 5 ml blood placed in EDTA tube and 10 ml in a plain tube, from all healthy volunteers one week before, on the third week of, and one month after the end of. All participants were chosen from the same living community, Rusaifa city, Jordan, so that the socioeconomic levels were highly similar. Anthropometric measurements that are measured for volunteers showed no significant differences between results before and after except for the body fat (%) in males (P=0.051). The packed cell volume shows significant reduction in males and females (P=0.052, P=0.04) respectively, while RBCs showed significant reduction in females only (P=0.01). white blood cells and the differential count showed no significant effect of fasting except for the monocyts absolute count as well as percentage in males which show significant increase(p<0.001). The results of the lipid profile showed an important significant increase in the concentration of high density lipoprotein cholesterol (P=0.006). Keywords fasting, Hematological parameters, HDL- Cholesterol, lipids profile. R I. INTRODUCTION AMADAN is a religious month during which all Muslims refrain from eating, drinking, and smoking during the daylight hours for a month. The duration of restricted food and beverage intake is approximately 12 h/day and changes depending on the geographical location (Aksungar, et al., Mohammad A. Fararjeh is Faculty of Medical Laboratory Technology, Hashemite University, Zarka, Jordan. Abdulrahim AlJamal is PhD, Associated professor, Faculty of Allied Medical Sciences, Department of Medical Technology, Zarqa Private University, P.O. Box Zarqa Jordan Work Phone Ext. 1814, Mobile +62-(0) E- Mail : aljamalzpu@yahoo.com "Mo'ez Al-Islam" E. Faris is in Department of Nutrition, Faculty of Pharmacy and Medical Sciences, Petra University. Ref'at A. Al-Kurd is in Department of Nutrition, Faculty of Pharmacy and Medical Sciences, Petra University. Mohammad Khalil is in Department of clinical pharmacy, Faculty of Pharmacy, University of Jordan.Jordan. Yasser Al-Bustanji is in Department of clinical pharmacy, Faculty of Pharmacy, University of Jordan.Jordan. 2005). During the fast, Muslims eat two meals a day, one before dawn and the other shortly after sunset (Haghdoost, and PoorRanjbar, 200). The fasting is considered a unique model of intermittent fasting (Aksungar, et al., 2005 ). As food and fluid intake became exclusive at nocturnal time without restriction on the type or amount of food intake. fasting as associated with alteration in meal frequency, sleep duration and reduction in physical activity during the day (El Ati, et al., 15). Intermittent fating is considered an alternative to caloric restriction regimen (Varady and Hellerstein, 2007), while other considered as a type of caloric restriction (Anson, et al., 2002). In general caloric restriction is define as restricting food intake to a level below that would be consumed voluntarily ( Anson, et al., 2002). Studies have shown that caloric restriction can increase life span, reduce the incidence of various age-related diseases and reduce oxidative stress and inflammation in rodents and non-human primates (Wan, et al., 2003). IF and other caloric restriction regimen has shown to reduce oxidative damage to lipids, protein, and DNA, which implies a protective effect against oxidative stress (Varady and Hellerstein, 2007).Two hypothesis are proposed to explain this beneficial effect: First, due to the reduction in overall energy intake, there is likely a reduction in reactive oxygen species (ROS) production and lower levels of lipid peroxidation products in the blood. Second, it may impose a mild repeated stress, to which cells respond adaptively by upregulating the expression of antioxidant systems, enhancing their ability to cope with more severe stresses and hence increasing disease resistance (Johnson, et al., 2007). The effect of intermittent fasting on oxidative stress in humans has been demonstrated in limited number of studies. Johnson, et al. (2007) have shown a significant decreases in serum level of 15-F 2t -IsoP in asthmatic over weight patients after 8 weeks of alternative day caloric restriction (ADCR). The decrease in levels of oxidative stress in subjects may have contributed to the improvement in their asthmatic symptoms on the ADCR diet. The quality of ingested nutrients can also differ during compared with the rest of the year, with a tendency to consume foods that are richer in carbohydrate and lipids 262

2 (Lamri-Senhadji, et al., 200). The daily energy intake during varies according to the food habits in different Islamic countries. In Saudi subjects for examples studies showed an increase of total energy intake, while Indian Moslems showed a decrease of daily energy intake during fasting (El Ati, et al., 15).Many physiological and psychological changes are observed during. It has been established that these changers occur at the end of the first week and adaptations require at least 10 days and remain the same for at least 10 days after (Aksungar, et al., 2005).The effect of fasting during on serum glucose showed reduction in serum glucose ceases due to increased gluconeognesis in the liver. This occurs due to the decrease in insulin secretion with the rise in glucagon and sympathetic activity during fasting state (Mansi, 2007). The effect of fasting on lipid profile is different in published articles and this may be due to a change in the dietary regimen during, decreased activity and some cultural parameters. fasting showed to have a beneficial effect on lipid profile by increasing HDL and decreasing LDL level (Ibrhim, et al., 2008; Lamri-Senhadji, et al., 200; Mansi, 2007). A significant changes observed during is the increase of serum uric acid, which could be explained by the increased intake of protein and fat during and the concomitant dehydration (El ati et al 15). Only few studies investigated the effect of on hematological parameters, (Unalacak M, et al. 2011).Our study will inspect the effect of fasting on the over all hematological parameters that could be considered as complete blood count (CBC) to evaluate the effect of this type of fasting on these important cellular as well as hematological parameters in healthy Jordanian population. Lipid profile will also be tested before, during, and after to study the effect of this type of intermitting fasting on these important biochemical parameters, in addition anthropometric measurements for the study subjects before, during, and after, will be conducted and subdivided according to sex. II. SUBJECTS Eighty healthy subjects (50 male and 30 female) volunteered to participate in this study. All were in the age range 18 to 60 were included. Volunteers who have diabetes, hypertension or metabolic disorders were excluded. None of the subjects was on any medication or took antioxidant. Female subjects were not pregnant or using contraceptives. This study was carried out in the month of of August/September 2010, and the average duration of fasting was hours a day. The mean climate temperature and humidity were 26.3 C and 44.6%, and 23.7 C and 55.8% during August and September respectively. There were no special nutritional regimens and recommendations during the whole study. To maintain sample homogeneity, all participants were chosen from the same living community, Rusaifa city, Jordan, so that the socioeconomic levels were highly similar. All participants were required to provide a written informed consent before the study started. A questionnaire was administered, in which medical history was recorded through a personal interview with the each subject by well-trained personnel. At the beginning of the study, the number of participants was 120, while 20 subjects decided not to undergo the second stage and another 20 subjects decided not to undergo the third stage. So that, the 80 remaining subjects did undergo all of three stages of the study. III. 2BLOOD SAMPLING PROTOCOL Blood samples were collected as follows, each volunteer at each visit gives 5 ml blood placed in EDTA tube and 10 ml in a plain tube, from all healthy volunteers one week before, on the third week of, and one month after the end of. anthropometric measurements (body weight, height, waist and hip circumferences), and body fat % were performed by well-trained personnel. Each subject served as his own control by comparing before values with those during and after. At each visit, systolic and diastolic blood pressures were measured, hip circumference HC, waist circumference WC and Body mass index BMI were measured with the subject in a seated position after 5 min rest. Blood sampling and processing was conducted in Al-Quds Medical Laboratories center between 10 am and 4 pm each time. The blood samples were collected and the serum was separated within an hour. IV. 3BMATERIALS AND METHODS The blood samples collected in EDTA tubes were determined the complete blood count (CBC) for each volunteer by using auto hematology analyzer (Mindray BC- 3000, Plus). While the blood samples in plain tubes were centrifuged at room temperature for 5 min. at 4000 rpm and serum was collected and pooled for analysis of lipid profile. Triglycerides were assayed using enzymatic methods (Roche Diagnostics, Mannheim, Germany). high-density lipoprotein cholesterol was measured enzymatically after precipitation of LDL and very low-density lipoprotein with Mg2+-dextran (Roche Diagnostics). LDL-c was calculated by Friedewald s formula. During each of the three visits, anthropometric measurements (body weight, height, waist and hip circumferences), and body fat % were performed by welltrained personnel. Body fat was measured by Body Fat Analyzer (GIMA, Italy). Body mass index (BMI) and waist to hip ratio were calculated. BMI was calculated as body weight (in kg) divided by body height (in meters) squared. Volunteers with BMI 25 Kg/m2 are considered to have normal weight, and those with BMI 25 are considered to be overweight (WHO, 2006). V. 4BRESULTS Table (1) shows that all the anthropometric measurements that are measured for the volunteers showed no significant 263

3 differences between results before and after except for the body fat (%) in males ( P=0.051). The table (2) shows the result of hematology parameters (complete blood count), the results were divided into two parts hemoglobin and RBCs indices and other for white blood cells and the differential counts percentage and as absolute counts. The comparison was between each parameter before and after fasting for both sexes. The packed cell volume shows significant reduction in males and females (P=0.052, P=0.04) respectively, while RBCs showed significant reduction in females only (P=0.01). The platelets showed no any significant changes. Table (2) also shows the total white blood cells and the differential count showed no significant effect of fasting except for the monocyts absolute count as well as percentage in males which show significant increase(p<0.001). The results of the lipid profile as indicated in the table (3) showed an important significant (P=0.006) increase in the concentration of the high density lipoprotein cholesterol HDL-c after fasting of compared to those before. While the total cholesterol, LDL-cholesterol and Triglycerides showed no any significant changes TABLE I DEPICTS THE ANTHROPOMETRIC CHARACTERISTICS OF STUDY SUBJECTS BEFORE, DURING, AND AFTER RAMADAN. Parameter Body weight (kg) BMI (kg/m2) Before MeanSD During MeanSD Male Female Male Female Male Female HC (cm) WC (cm) Body fat (%) HC: hip circumference, WC: waist circumference, BMI: Body mass index CBC Parameter After MeanSD a P b P P 0.05 is considered to be statistically significant. TABLE II THE HEMATOLOGICAL PARAMETERS RESULTS FOR THE MALES AND FEMALES BEFORE, DURING, AND AFTER RAMADAN. Before MeanSD During MeanSD Male Female Male Female Male After MeanSD a P Female Hb( g/dl) PCV(%) RBCs (x10p P/L) MCV (fl) MCH(pg) MCHC (g/dl) RDW b P 264

4 CV(%) RDW SD(fL) Platelets (x10p P/L) MPV(fL) WBCs (10P P/L) Lymph# (x10p P/L) Mono# <0.00 (x10p P/L) Gran# (x10p P/L) Lymph (%) Mono(%) < Gran(%) Hb: Hemoglobin; PCV: Packed cell volume; RBCs: Red blood cells; MCV: Mean corpuscular volume; MCH; Mean corpuscular hemoglobin; MCHC: Mean corpuscular hemoglobin concentration;rdw: Red blood cell distribution width; MPV: Mean platelet volume (p <0.05), * Significant difference TABLE III LIPID PROFILE CONCENTRATION RESULTS FOR THE MALES AND FEMALES BEFORE, DURING, AND AFTER RAMADAN. Biochemical test Before During After P Total cholesterol HDL-Cholesterol LDL-Cholesterol Triglycerides VI. 5BDISCUSSION The anthropometric measurements that are measured for the volunteers showed no significant differences between results before and after except for the body fat (%) in males. These results are in concordance with (Heilbronn, et al., 2005) founded that Alternative day fasting is usually associated with weight loss and increased fat oxidation which is translated to fat mass loss. While (Haghdoost, and PoorRanjbar, 200) shows more foods rich in fat and dietary cholesterol were consumed during this period. The effect of fasting on body weight. Despite the marked changes in food intake habits some studies showed that fasting had no effect on body weight or BMI (El ati, et al., 15; Aksungar, et al., 2005; Lamri-Senhadji, et al., 200; Ibrhim, et al., 2008), while other studies showed that fasting was associated with significant weight loss (Ziaee, et al., 2006; Mansi, 2007; Al-Hourani and Atoum, 2007). These studies agree with our study. The hematological parameters (complete blood count), results were divided to hemoglobin and RBCs indices while the second for white blood cells and the differential counts percentage. The packed cell volume shows significant reduction in both sexes, while RBCs showed significant reduction in females only. The significant reduction in the PCV and RBCs count may because of the disturbance in the body homeostasis in volunteers during and after fasting, (Unalacak et. al. 2011). The table also showed that the total white blood cells and the differential count showed no significant effect of fasting except for the monocyts in males which shows significant increase in their number this may because the fasting stimulate some cytokine production which stimulate the production of the monocytes. The results of the lipid profile shows an important significant increase in the concentration of the high density lipoprotein cholesterol. These promising results are with concordance with the results of Mansi et. al 2007 that showed increased in HDL- 265

5 cholesterol levels, suggest a beneficial effect on coronary heart disease (CHD) patients. As a matter of fact, Temizhan and colleagues (1) have evaluated the effect of fasting on a population of CHD patients. They found that the number of cases with acute CHD events was lower in, thus confirming the beneficial effect of fasting in such population. While other studies showed no significant changes in TG and LDL levels with significant elevation in HDL levels (Aksungar, et al., 2005).These founding agree with our study. Heilbronn, et al. (2006) showed no significant change in level of protein carbonyl and a significant decrease in DNA damage after 6 month of caloric restriction in healthy volunteers. These findings should be taken into account when evaluating the effects of on the general population. To avoid the adverse effects of anemia, increased intake of iron-rich foodstuffs is recommended during the month, regardless of fasting practice. Farther investigations concerning the effect of the fasting on the function of the immune cells including the monocytes, type of cytokine stimulation and the mechanism of increasing the high density lipoprotein cholesterol are also recommended. Inflammation in Overweight Adults With Moderate Asthma. Free Medical Biology and Medicine, 42(5), [10] Lamri-Senhadji, M. Y. El Kebir, B. Belleville, J. and Bouchenak, M. (200), Assessment of Dietary Consumption and Time-Course of Changes in Serum Lipids and Lipoproteins Before, During and After in Young Algerian Adults. Singapore Medical Journal, 50(3) [11] Mansi, K. M. S. (2007), Study the Effects of Fasting on Serum Glucose and Lipid Profile among Healthy Jordanian Students. American Journal of Applied Sciences 4(8), [12] Temizhan, A. Donderici, O. Ouz, O. and Demirbas, B. (1), Is There any Effect of Fasting on Acute Coronary Heart Disease Events? International Journal Of Cardiology, 70(2), [13] Unalacak, M. Kara, I. H. Baltaci, D. Erdem, O. and Bucaktepe, P. G. (2011), Effects of Fasting on Biochemical and Hematological Parameters and Cytokines in Healthy and Obese Individuals. Metabolic Syndrome and Related Disorders. [14] Varady, K. A. Roohk, D. J. and Hellerstein, M. K. (2007), Dose Effects of Modified Alternate-Day Fasting Regimens on In Vivo Cell Proliferation and Plasma Insulin-Like Growth Factor-1 in Mice. Journal of Applied Physiology, 103(2), [15] Wan, R. Ismail, A. Brown, M. Cheng, A. Kamimura, N. Talan, M. and Mattson, M. P. (2010), Cardioprotective Effect of Intermittent Fasting is Associated With an Elevation of Adiponectin Levels in Rats. Journal of Nutritional Biochemistry, 21(5), [16] Ziaee, V. Razaei, M. Ahmadinejad, Z. Shaikh, H. Yousefi, R. Yarmohammadi, L. Bozorgi, F. and Behjati, M. J. (2006), The Changes of Metabolic Profile and Weight During Fasting. 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