IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VO L.24 (3) 2011 Serumleptin, Triglycerides, Cholestrol and Hdl- Cholestrol In Non Dlabetic Obese Subjects M.A. Kassim Department of Biology, College of Science ،University of Al-Mustansyria Received in : 16 February 2011 Accepted in: 16 June 2011 Abstract This study was carried out to investigate the relat ionship between Serum leptin, Body Mass Index (BMI), T riglycerides, Cholesterol and HDL- Cholesterol in Non Diabetic Obese subjects comparing with healthy subject . A 36 male and female Iraqis obese were studied, mean age 50.1, 43.3 years respectively and 23 healthy subjects. Serum leptin, T riglycerides, Cholesterol and HDL- Cholesterol were measured. Leptin, T riglycerides, Cholesterol, HDL- Cholesterol and BMI significantly increased in obese males and females compared with control, but there was no significant difference in HDL- Cholesterol and BMI when compared between obese males and females. A low significant positive correlat ion was found between leptin and HDL- Cholesterol, and no significant difference between leptin and other types of lipids. The findings suggested that serum leptin concentrat ion topmost in obese subjects with noticeable effect on lipid metabolism. Key word: Serum leptin, Obesity, lipids Introduction Obesity is increasing progressively worldwide, It is closely associated with increased morbidity and mortality caused by several of the most common diseases in the world including gallstone diseases and cancer [1]. Obesity is determined by an interaction between environmental, psychosocial, and genetic factors [2]. Leptin (from the Greek word leptose, meaning thin) is a protein hormone that is encoded by the Ob gene and is secreted by adipose t issue to in the circulation. It acts mainly in the hypothalamus by binding to specific Leptin receptor and regulates food intake and energy balance [3]. In addition Leptin act ivates sympathetic nervous system and increases energy expenditure. Leptin thus decreases body weight and adiposity as a novel messenger of energy metabolism [4]. The information concerning the relationship between serum Leptin concentration and levels of serum lipids is important. Furthermore, when evaluat ing Leptin levels and metabolic profiles of patients with and without ischemic heart disease, no differences in Leptin concentration and no relat ionships between Leptin and plasma lipid were observed [5]. However, serum Leptin concentration in individuals with first- ever hemorrhagic stroke was sharply higher than in controls suggesting some association between serum Leptin and alteration of lipid metabolism [6]. Therefore, the aim of the present study is to evaluate serum Leptin, triglycerides, cholesterol and HDL- cholesterol in Iraqis obese people comparing with normal subjects. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VO L.24 (3) 2011 Materials and Methods Subjects A tot al of 36 (16 male and 20 female) Iraqis obese (BMI ≥ 30 Kg / m2) were enrolled in this study. Median age of the 36 female/male volunt eers was 43.0, 50.1 years respect ively, ranged (20 -61),(20 - 78)years respect ively. T hese out-patients were turned up t o AL-Numan Hospital laboratory for check-up for the period from June to August 2010 .A total of 23 control subjects(11 male and 12 female) with (BMI ≤ 25 Kg / m2) were also enrolled in this study. Both test and control groups were non diabet ic (serum glucose concentrat ion ≤110) and with normal blood pressure. BMI was calculated as weight (in kg) divided by squared height (in m2). Sample Collections Five milliliters (5ml) venous blood was obtained between 08:00 and 10.00 a.m. aft er a 12 hour fasting period. All blood samples were dispensed into dry glass test tubes for clotting and ret raction to take place. Sera were obtained aft er samples were centrifuged at 2000g for five minutes and stored at -20°C until assayed for laboratory investigat ions [7]. Laboratory techniques Fasting serum glucose concentrat ions had been measured by t he glucose oxidase procedure and serum HDL-cholesterol, t riglycerides, cholesterol were measured enzymetically by quantitat ive determination kit (SPINREACT, S.A.U. SPAIN). Stored frozen sera samples were retrieved, thawed, and tested for Leptin levels by means of a quantitat ive enzyme immunoassay using a commercial kit (DRG Inst ruments GmbH, Germany) (specificity, 100%). Reference standards were used to produce a standard curve to quantitate Leptin levels. T he results were expressed in ng/ml. T he normal value for Leptin levels was up 3.84±1.79 ng/ml in male and 7.36±3.73 ng/ml in female. Statistical analysis The stat istical computer software package MINIT AB was used to analyze t he data. The mean, range, and 95% confidence interval (C.I.) for median, were calculated. T he relat ionships between the individual parameters were evaluated using Spearman’s correlat ion. P-value < 0.05 was considered stat istically significant. Results and Discussion The study included 36 obese and 23 control subjects were divided in two groups, males and females. Mean, range and 95% confidence interval (CI) for age, serum Leptin, HDL, T riglyceride, Cholesterol, Glucose and BMI of obese females and control females are shown in table (1). The mean ages of the obese females and control females were 43.3 years (20-61) and 30 years (21-28), respectively. All of the parameters in table (1) were significantly higher than those of the control except glucose. Conversely, HDL and BMI were significantly lower between obese females and control females. The same parameters were applied to obese males and control males shown in table (2), the mean ages of obese males 50.1 years (20-78) and 35.25 years (24-43), respectively. Each of the parameters in table (2) was significantly high except glucose. In table (3) the triglyceride and cholesterol showed high significant increase in obese females compared with males, while serum leptin showed low significant. Conversely, HDL, glucose, BMI, and age showed non significant change between obese males and obese females. The relationships between the studied parameters in the obese males and females group are shown in table (4). As expected a highly significant positive correlation was found between BMI and serum leptin. In addition the table demonstrated a low significant positive correlation between leptin and HDL. Furthermore no significant correlation between leptin and cholesterol, IBN AL- HAITHAM J. FO R PURE & APPL. SC I. VO L.24 (3) 2011 triglyceride was foun . Also there was no significant correlation between BMI and cholesterol, triglyceride, HDL. In humans, it is well established that plasma leptin levels are directly proport ional to percentage body fat . Most obese individuals have high concentrat ions of leptin in their serum and plasma but exhibit leptin resistance because of decrease leptin transport into the central nervous system or down regulation of leptin receptors [8, 9]. In our study higher leptin concentrations were found in Iraqis obese males and females (BMI ≥ 25.00kg/m2) than the normal control group. The difference between the median of leptin in both obese males and females was significant depending on the distribution and composition of fat in the body or depending on the effect of sex steroids like estrogen and progesterone. [10, 11, 12] In the subjects investigated in the present study, the relationship between leptin and BMI was linear, as shown in table 4. The results similarly results presented in this study in good agreement with result reported in Saudian overweight and obese females in Makkah community [13], where significant relationship was found between leptin, BMI and waist circumference. Also t he results seemed to agree with those reported by Chaisiri et al. in Japanese obese women [14], where a linear relationship between serum leptin and BMI was found. On the other hand, the relationship between leptin and BMI in Caucasian individuals is not linear in very obese persons because Caucasians have different metabolic states from Asians, in addition for t he differences might be the greater height of Caucasians, so that an increase in the volume of fat t issue to a given height does not correspond in the same way as in Asians. The increase in fat t issue in Asians, with a relatively shorter height, results in a more direct relationship between fat t issue and BMI. It was found that blood-brain barrier transportation had a threshold level for serum leptin (about 25-30 ng/ml), above which increases in serum levels were not translated into proportional increases in cerebrospinal or brain leptin levels, which means t hat it may result in apparent leptin resistance and obesity[8]. Most investigat ions have demonstrated, in agreement with the data of this study, a lack of correlation between serum leptin and lipid profiles [15,16,17,18,19] mainly when body weight and degree of adiposity are taken into account References 1.Mantzoros,C.S . ،)1999 ( The role of leptin in human obesity and disease. A review of current evidence. Ann Intern. Med, 130:651-657 2.Kopelman, P .G. . ،)2000 ( Obesity as a medical problem. Nature,404:635-643. 3.Ahima, R.S. and Flier, J.S .،)2000 ( Leptin. Ann. Rev. Phys.62:413-437. 4. Sagawa N., Yura S., Itoh H., MIse H., Kakui K. Korita D ) .2002 ( Role of leptin in pregnancy-A review. Placenta:16 :S80-86. 5. Sodenberg, S.; Ahren, B.; Jansson, J.H ) .1999 ( Leptin is associated with increased risk of myocardial infraction. J Int. Med.246:409-418. 6.Couillard, C.; Mauriege, p.and Prud hommer, D. ) .1997 ( Plasma leptin concentrat ion: gender differences and associat ion with metabolic risk factors for cardiovascular disease. Diabetologia ؛ 40:1178-1184. 7. Alan, H.B. ) .2006 ( T ietz Clinical Guide to Laboratory T est ،4 th ed WB Saunders, Philadelphia. 8.Caro, J.F.; Kolaczynski, J.W.and Nyce, M.R. ).1996 ( Decreased cerebrospinal fluid serum leptin ration in obesity: A possible mechanism for leptin resistance .Lancet ,348:159-161. 9.Bjorbaek, C.; El Haschimi, K.; Frant z, J.D.and Flier, J.S. ) .1999 ( The role of SOCS -3 in leptin signaling and leptin resistance. J Biol. Chem.274:30059-30065. IBN AL- HAITHAM J. FO R PURE & APPL. SC I. VO L.24 (3) 2011 10.Fluda, S.; Linseies, J.; Wolfram, G.; Himmerch, S.; Gedrich; K., Pollmacher. T .and Himmerich, H. . )2010 ( Leptin plasma levels in the general population influence of age, gender, body weight and medical history. P rotein Pept. Let t .17(11):1436-1440. 11.Minoccin, A.; Savia, G.; Lucantoni, R.; Berselli, M.E.; Tagliaferri, M.and Calo, G.and Petroni, M.L. ).2000 ( Leptin plasma concentrat ion are depend on body fat distribution in obese patients. Int. J. Obese Relat . Metab. Disord24. (9): 1139 - 1144 12.Steven, M.; Heikki, M.; Pauli, K.; Leena, M.and Markku, L. ).1997 ( Leptin concentrat ion sex hormones, and cort isol in non diabet ic man. J. Clin. Endocrinol and Metab .82(6):1807-1813. 13.Bahathiq, A.O. ).2010 ( relationship of leptin hormones with Body Mass Index and waist circumference in Saudi female population of the Makkah community. The Open Obesity J.2:95-100. 14.Chaisiri, K.; Pongpaew, P.; Tungtrongchitr, R.; Phonrat , B.; Kullrap, S. and Schelp, F.P ) .1998 ( Nutritional status and serum lipids of a rural population in Northeast Thailandan example of health transition. Int. J. Vitam. Nutr. Res.68:196-202. 15.De courten, M.; Zimmet, P.and Hodge, A. ) .1997 ( Hyperlept inaemia: T he missing link in the metabolic syndrome? Diabet. Med.14:200-208. 16.Haluzik, M.; Fiedler, J.and Nedvidkova, J. ) .2000 ( Serum leptin levels in patients with hyperlipidemias. Nutrit ion. 16:429-433. 17.Leyva, F.; Godsland, I.F.and Ghatei, M. ) .1998 ( Hyperleptinemia as acomponent of metabolic syndrome of cardiovascular risk. Arterioscler T hromb Vasc. Biol.18:928-933. 18.Ryan,A.S.and Nicklas, B.J. ) .1999 ( Age-related changes in fat deposition in mid-thigh muscle in women: relat ionships with metabolic cardiovascular disease risk factors. Int. J. Obes126- 23 :-132. 19. Ho, S.C. ;T ai, E.S.and Eng, P.H.K. (1999) A study in the relat ionship between leptin, insulin and body fat in Asian subject . Int . J. Obes.; 23: 246-252. Table : )1 ( Mean, ranges an d 95% CI of age, leptin, HDL, Triglyceride, Cholesterol, Glucose and BMI in obese and control females Control Female O bese Female Normal value P- value 95 %CI Mean (range ) 95 %CI Mean (range) Parame ters 0.00 21.6-23.8 30 (21-28) 33.5-41.81 43.3(20-61) Age yrs 20-25 0.05 22.3-23.1 24.3(21.5-25.3) 26.3-30.8 31.27(30-34.6) BMI Kg/m2 7.3 0.00 0.4-1.2 2.8(1.8-7) 8.3-11.6 15.83(1.2-50.7) Leptin mg/ml 40-140 0.00 82.4-110.9 106.1(79.6-132.7) 141.2-156.8 172.78(70.7-265) Triglyce ride mg/dl 250-140 0.00 115.7-130.8 138.8(108-154.5) 196.3-218.2 198.47(69.5-243) Cholest erol mg/dl 72-40 0.01 46.3-526 57.9(54-61.7) 41.3-48.6 46.49(38.6-61.7) HDL mg/dl 70-110 N.S. 75.6-81.7 79.2(72-84.6) 75.4-88.6 83.35(55.8-110) Glucose mg/dl BMI = body mass index, HDL-C= high density lipoprotein cholesterol, Significant difference between obese Female and control Female subjects p ≥0.05 IBN AL- HAITHAM J. FO R PURE & APPL. SC I. VO L.24 (3) 2011 Table (2): Mean, ranges and 95%CI of age, leptin, HDL, Triglyceride, Cholesterol, Glucose and BMI in obese and control males Control male Male Obese Normal value P-value 95 % CI Mean (range ) 95 % CI Mean (range( Paramet ers 0.01 39- 6 .28 .1 35-25(24-43) 6 .51- 6 .33 )78 -20 (1 .50 Age yrs 25-20 0.05 1 .23- 1 .21 )25.3- 5 . 21( 24 1 .32 -6 .29 )1 .38 -30 (7 .32 BMI Kg/m2 3.8 0.00 1.1 -37 .0 1.7(1-4) 6 .10- 9 .6 )1 .44 -3 .1 (4. 17 Leptin mg/ml 160-60 0.00 198- 3 .95 )106 - 106 (106 1 .163 – 6 .128 136.(70.7-273) Triglycer ide mg/dl 140-250 0.00 118.3- 78 .91 )131 - 9 .84( 108 6 .191 – 3 .146 169.5(96.5-243.2) Choleste rol mg/dl 40-72 0.00 54.81-59.1 )7 .61 -65 (35 .63 7 .51 – 13 .43 49.9(34.7-65.6) HDL mg/dl 70-110 N.S. 68.6-81.33 82.8(75.6-90) 63 .88 – 3 .85 93.9(72-113.6) Glucose mg/dl BMI = body mass index, HDL-C= high density lipoprotein cholesterol, Significant difference between obese male and control male subjects p ≤ 0.05 Table (3) Mean, ranges and 95%CI of age, leptin, HDL, Triglyceride, Cholesterol, Glucose and BMI in O bese males and females Obese female Obese male P-value 95 %CI Mean (range( 95 %CI Mean (range( Parameters N.S. 33.5-41.81 43.3(20-61) 33.6-51.6 50.1(20-78) Age yrs N.S. 26.3-30.8 31.27(30.65) 29.6-32.1 32.7(30-38.1) BMI Kg/m2 0.05 8.3-11.6 15.83(1.2-50.7) 6.9-10.6 17.4(1.3-44.1) Leptin mg/ml 0.001 156.8 -141.2 . 172.78(70.7-265) 128.6-163.1 136.9(70.7-274) Triglyceride mg/dl 0.00 2218. -3 .196 198.47(89.5-243) 146.3-191.6 169.5(96.5-243.3) Cholesterol mg/dl N.S. 41.3-48.6 46.49(38.6-16.7) 43.13-51.7 49.9(34.7-65.6) HDL mg/dl N.S. 75.4-88.6 83.35(55.8-110) 85.3-88.63 93.9(72-113.5) Glucose mg/dl BMI = body mass index, HDL-C= high density lipoprotein cholesterol, Significant difference between obese male and control female subjects p ≤ 0.05 Table (4) Correlation coefficients of age, leptin, HDL, Triglyceride, Cholesterol and BMI in obese males and females Age BMI HDL Triglyceride Cholestero l Leptin Parameters 0.361** 0.710** 0.205* -0.059 -0.029 1.00 Leptin 0.338** -0.043 0.204** 0.285** 1.00 -0.029 Cholesterol 0.201** - 0.041 -0.410** 1.00 0.285** -0.059 Triglyceride 0.244** -0.038 1.00 0.140** 0.204** 0.205* HDL 0.722 1.00 -0.038 0.041 -0.043 0.710** BMI 1.00 0.722 0.244** -0.201** 0.338** -0.361** Age Significant difference: *p <0.05,**p<0.01 2011) 3( 24مجلة ابن الهیثم للعلوم الصرفة والتطبیقیة المجلد نیه عالیة الكثافة الكولسترول و البروتینات الده، الدهون الثالثیة ، هرمون اللبتین في مصل دم البدناء غیر المصابین بالسكر مهند عبد اإلله قاسم الجامعة ألمستنصریه،كلیة العلوم ، قسم علوم الحیاة 2011شباط 16:استلم البحث في 2011 حزیران 16: قبل البحث في الخالصة یــة ، ) BMI(مؤشــر كتلـة الجســم ، رمــون اللبتـین تـم التحـري فــي هـذه الدراســة عـن العالقــة بـین ه الكولسـترول فضــآل ، الـدهون الثالث ابین بمـرض السـكر مقارنـة باألصـحاء شخصـآ مـن البـدناء 36شـملت الدراسـة . عن البروتینات الدهنیة عالیـة الكثافـة لـدى البـدناء غیـر المصـ ناثا لهم معـدل عمـر تـوا 43,3: 50,1العراقیین ذكورا و إ قـیس تركیـز هرمـون اللبتـین و نسـبة الـدهون . عینـة لألصـحاء 23لي و سـنة علـى ال افــة ،وشـملت الكولســترول یــة الكث نــات الدهنیـه عال ـة، و البروتی تــین ، و الــدهون الثالثیـ ـادة بتركیــز هرمــون اللب ــوي وزیـ ــوحظ وجــود فـرق معن ، ل افــة اء الـذكور و اإلنـاث مقارنــة BMIو الكولسـترول، و الـدهون الثالثیـة فضــآل عـن،البروتینـات الدهنیـه عالیـة الكث عنـد عینـات الدراســة للبـدن یــة الكثافـة، و ، باألصـحاء اء مــن BMIلكـن لـم یالحـظ وجــود فـرق معنـوي للبروتینـات الدهنیـة عال اء الـذكور مـع البـدن بــدن عنـد المقارنـة بـین ال ابي قلیـل فـي العالقـة بـین هرمـون اللبتـین و البروتینـ. اإلناث ات الدهنیـه عالیـة الكثافـة بینمـا لـم یالحـظ فـرق معنـوي كما وجد فرق معنـوي ایجـ اء مقارنة باألصـحاء . بین هرمون البتین و بقیة أنواع الدهون من تلك النتائج وجد أن هرمون اللبتین كان بمستوى عال عند األشخاص البدن .ولم یكن له تأثیر ملحوظ في التكوین األیضي للدهون لكلمات المفتاحیة الدھون , البدانة , ھرمون اللبتین : ا Age