IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 Association Between Lipid Profile ,BMI , and Some Pituitary Hormones Abnormalities In Sera of Iraqi Infertile Females A. R.Mahmood Department of Chemistry, College of Education Ibn – Al-Haitham, University of Baghdad Abstract In this study , the clinical impact of interaction between gonadotrophin hormones (luteinzing hormone, LH, and follicle stimulating hormone ,FSH ) and prolactin PRL in serum of seventeen Iraqi infertile female with the lipid profile . In addition to control group involving age matched fertile females . Immunoradiometric assay ( IRMA ) technique for the determination of (LH , FSH and PRL) was utilized. The lipid profile { i.e. total cholesterol ( Tc ) , triglycerides (TG) , and high density lipoprotein – Cholestrol ( HDLc )} ,were evaluated by using colorimetric method , while{ low density lipoprotein cholesterol ( LDLc ) and very low density lipoprotein – cholesterol ( VLDLc )} , were evaluated by using a mathematical formulas. The body mass index ( BMI ) was calculated as weight ( Kg) / height ( m² ). The results revealed that only( 20%) of infertile female with abnormal hormonal levels are obese , while (40% )of them were overweight . Only (TG ) in sera of infertile female showed a significant increase than that of control group. On the other hand no significant differences in (Tc) , (HDLc) were noticed in sera of groups under consideration . A conclusion could be obtained from the above data , that (BMI )or obesity is not associated with infertility always. Introduction The reproductive system of women shows regular cyclic changes that may be regarded as periodic preparations for fertilization and pregnancy . This cycle is the menstrual cycle [1] . Menstruation is dependent on the proper functioning of a chain made up of ; hypothalamus → pituitary →ovary(HPO) ; amenorrhea presupposes awakening or break in one or more of these links [2] . Interaction ( HPO ) to produce cyclic expression of the appropriate hormones at the expected time or chronology age at which sexual maturation should normally occur [3] The growth and the reproductive activities of the gonadal tissue are controlled by the gonadotrophins hormone LH and FSH from the interior pituitary gland . LH and FSH are called gonadotrophins because they regulate the function of the gonads (ovaries and testes ) , in both sexes , FSH stimulates gamete ( sperm or egg ) production, while LH promotes production of gonadal hormones [4] .The secretion of both LH and FSH is stimulated by gonadotrophin releasing hormone from the hypothalamus , LH and FSH both are subjected to feed back loops regulation by the ovarian hormones [5]. In women , LH along with FSH are ordered as part of the workup of infertility and also useful in the investigation of menstrual irregularities , and to aid in the diagnosis delayed and precocious puberty [6]. Prolactin PRL stimulates mammary development and subsequent lactation , it is an episodic secretion that it p roduced by the lactotrophs of interior pituitary[7] . IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 High level of PRL is a biochemical finding and does not necessarily indicate the presence of a disease , this would increase in PRL level in the blood above the physiological concentration, it is now well documented to interfere with the female reproductive function [8] . It was reported that obesity was a common finding in woman with ovarian hyperandrogenism , although the mechanisms underlying this relationship remain largely undetermined [ 9 ]. The aim of the present study is to evaluate the lipid profile of infertile woman with pituitary hormones dysfunction and compared with that for healthy fertile woman matched in age and body mass index BMI. Experimental Part Seventeen infertile female were selected from Elwyia Maternity Teaching Hospital during 2008, a careful history was obtained from the patients including menstrual disturbance , if any , associated with the symptom of infertility , weight and height and all patients, were free from medication affecting hormone level . Evaluation of each patient is done by detecting body mass index, serum gonadotrophens hormones level LH , FSH and serum PRL. Excluding the test during the days (12 – 16)from the cycle period . As control (17) normal female with normal regular menstrual cycles with body mass index within normal range ,patients and control aged ( 20 – 39 ) years were included in this study . Blood samples ( 5ml ) were collected in plain tubes , centrifuged at 3000 rpm for 10 min, after allowing the blood to clot at room temperature . The sera were a liquated and frozen until assay performed . Body mass index uses a mathematical formula based on person's height and weight , BMI equals weight in kilograms divided by height in square meters ( BMI = kg / m²) [10]. Fost et al suggested that a BMI of ( 18.5 – 24.9 ) indicates a person of normal weight . A person with a BMI of ( 25 – 29.9 ) is overweight , while a person with a(BMI of ≥ 30) is obese [11] . Immunoradiometric assay IRMA for the in vitro determination of FSH , LH , and PRL in human serum was utilized by kits from immunotech abeckman coulter company . The Immunoradiometric assay of luteinizing hormone FSH , LH , and PRL is a sandwich type assay the kit utilizes mouse monoclonal antibodies directed against two different epitopes of FSH , LH and PRL hence not competing . Samples or calibrators are incubated in tubes , coated with the first monoclonal antibody in the presence of the second monoclonal antibody labeled with iodine 125. The content of tubes is aspirated and rinsed after incubation and bound radioactivity measured values are calculated by interpolation from the standard curve . The radioactivity bound is directly proportional to the concentration of FSH , LH , and PRL in the samples. The results are obtained from the standard curve by interpolation. The standard curve serves for the determination of FSH , LH and PRL concentration in samples measured at the same time as the calibrator . Determination of total serum cholesterol [12] involves the use of three enzymes , cholesterol esterase , cholesterol oxidase and peroxidase . In the presence of the former mixture, ( N- ethy l propyl – m – anisidine ) and 4 – amino – antipyrine are condensed by hydrogen peroxide to form quinoneimine dye proportional to the concentration of cholesterol , when the absorbance of the samples measured against the reagent blank within 60 minutes at 500 nm. IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 The Triglycerides were determined after enzymatic hydrolysis with lipases . The indicator is aquinoneimine formed from hydrogen peroxide , 4-aminophenazone , and 4 – chlorophenol under the catalytic influence of perioxidase . The absorbance was measured for test and standard against the reagent blank within 60 minutes at 500 nm [13]. In determination of high density lipoprotein – cholesterol HDLc [14], the method uses a selective precipitations of chylomicrones and the apolipoprotein containing lipoprotein VLDLc and LDLc by addition of 4% phosphotungstic acid solution , which contain 10 % magnesium chloride PH 6.2 . Sedimentation of the precipitant by centrifugation , and subsequent enzymatic analysis of HDLc as residual cholesterol remaining in the clear supernatant , from which the cholesterol can be determined as described as above according to [12]. Low density lipoprotein cholesterol LDLc was determined by using empirical Friedwald formula which was based on the assumption that VLDLc is present in serum at concentration equal to one fifth of the TG concentration . This formula is as follow [15] :when all concentration are given in milligrams per deciliter :- LDLc(mg / dl) = Total cholesterol - ( HDLc +VLDLc ) Statistical Analysis of Data To compare the significance of the differences in the mean values of any two groups student's t-test was applied and P value less than 0.05( p<0.05) was considered statistically significant . Results and Discussions Table (1)and fig(1) showed the mean distribution of serum hormones concentration LH , FSH , and PRL of the infertile female patients with the minimum and maximum values to each hormonal level when compared with the mean concentration of the normal control . A significant difference was observed between normal and high hormonal levels of infertile female compared to the majority of infertile have hormonal levels higher than that of control with a percentage of 74.4%, on the other hand there was no significant differences in the level of FSH in 28% of infertile female compared with healthy control . The significant difference was found in the higher levels of FSH in 72% of infertile female compared with normal hormonal level of healthy control . The results agreed partly with other studies who concluded that different results could reflect variation in the selection of patients and / or the different lifestyle factors [16]. The mean serum level of LH / FSH ratio for patients was(0.8 )and for control group was(0.8). The difference is not significant yet others studies reported higher or lower ratio and they stated that this variation might be due either to primary central disorders involving (GnRH) secretion or secondary pituitary sensitization to (GnRH) by an abnormal feed back signals from ovaries [17] . The PRL level in (60%) of infertile female showed a significant increase compared to that of healthy control while lower significant values than control was depicted in the rest of the patient .High prolactin level are found in( 30% )of women with different kind of amenorrhea leading to infertility [18] . About( 60% )of patients of high level of prolactin showed lower levels of FSH and LH than healthy control. These results are in agreement with other studies who suggested that a decline in gonadotrophine in hyperproctanemic patient showed the association between gonadatrophine deficiency and hyperprolactinemia which might be an indirect sign of functional hypothalamic pituitary interruption due to the inhibitory effect of PRL [19] . Body mass index is a measurement that is associated with body fat and it is widely used by health care provider. IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 Table (2)represent mean±SD of BMI for both groups .Fig (2) showed the distribution of infertile female according to BMI.There was a significant difference between patients, patients were overweight (40 % ), and( 33.3 %) of patients were in a healthy weight range ,while the percentage of obese patient was( 20%). The present results disagree with some reports of infertility association with obesity [20 ] . Table (3) and fig (3) showed the lipid profile in serum of the infertile female and the healthy control. The mean serum level of total cholesterol Tc for the infertile group was (172.125± 35.58 mg /dl ) and for the control group was (171± 14.83mg / dl ). The values for triglycerides TG were (108.68 ±31.63 mg /dl) and (80 ±7.19mg /dl )for the infertile females and control respectively . The HDLc and LDLc were (57.75 ±7.56 mg / dl) and (97.68 ±27.14 mg / dl) in serum of the infertile female respectively , and were (54.7± 7.39mg/dl) and (100.1 ±10.38mg /dl )for the control group respectively . The values for VLDLc were (21.68±6.33mg/dl)and(16±1.438mg/dl) for the infertile females and control respectively .A non significant elevation in total cholesterol as compared with control group was found .The result does not agree with other studies which found that women with different causes of infertility had increase Tc levels[21]. A significant elevation in TG in serum of infertility female patient compared to control was obvious as shown in table( 3 ). The pattern of dyslipidemia is mostly found in a wide range of causes of infertility , even studies reported that high TG levels are found in both obese and non obese women suffering from infertility [22&23 ] . Abnormalities of LDLc had not been found consistently in some cases of infertility , even in those with a normal LDLc level had shown increase VLDLc and small dense LDLc relative to control subject [24] . High TG and low HDLc in patients with hyperprolactinemia were reported due to acceleration of hepatic and adipose tissue biosynthesis of triglyceride and certain phospholipids factors [ 25 ]. References 1.Ganong, W.F. ( 1999 ) .The Gonads Development and Function of the Reproductive System In : A review of Medical Physiology ( 19 th edition) . Appleton and Lang Medical Publishment – Stamford , Connecticut .P 393 – 432. 2.Tindall ,V.R. ( 1987 ) . Amenorrhoe : Scanty and Infrequent menstruation . In : Jeffcoate's principles of Gynaecology , ( 5 th edition ) . Butterworth & Co. ( Publishers ) Ltd. P.495 – 496 . 3.James , D. and Brian , A.M. ( 2004 ) " Amenorrhea " .In " Clinical obstetrics and Gynaecology " , ( 1st edition ) . Saunders . P 127 . 4.Elaine , N. and Marieb , R.N. ( 2000 ) : The Endocrine System in Anatomy and physiology . Benjamin Cummings , San Francisco , Chapter 15: 512 , 518 . 5.Zilva , F. ; Mayne , D. Philip and Pannel ( 1999 ) The hypothalamus and pituitary gland .In : Clinical chemistry in Diagnosis and Treatment , 6th ed. Hodder Headline group PLC London , chapter 5: 106 – 115. 6.Mc Donough ,P.G. ( 2003 ) : Molecular abnormalities of FSH and LH action , Ann N Y Acad Sci 997 : 22 – 34. 7.Marc , E.F.; Bela , k. ; Anna ,L. and Gyorgy , N. ( 2000 ) , p rolactin : Structure , Function and regulation of section : Physiological Reviews 80 ( 4 ) : 1523 – 1631 8.Hanan , L.S. ( 2002 ) . The effect of hyperprolactinema on Follicular development and estradiol concentration in stimulated cycle .M Sc. Thesis , college of medicine . Univ. of Baghdad , Iraq .P.44 – 46 . 9.Franks , S. ; Kiddy , D. and Sharp , P. ( 1991 ) obesity and polycystic ovary syndrome . Ann N.Y. ,Acad.Sci . 626 : 201 – 206 . IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 10.Dennis , L.K. ; Eugene , B. ; Anthong , S.F. ; Stephen , H. and Dan, L.L. ( 2005 ) Harrison 's principles of Internal Medicine , Volume 1, ( 16th edition ) . McGraw Hill ,Medical Publishing Division. P. 423 – 425 . 11.Fost , L. ; Hane , L.J. ; and Vestergaard, . p.( 2005) " Over weight and obesity as risk factors for a trial fibrillation or flutter The Danish diet ,Cancer , and health study " Am. J. of 17ed , 118 : 489 – 95 . 12.Richmond , W. ( 1974 ) Proceedings in the development of an enzymatic technique for the assay of cholesterol in biological fluids .Clin Scc Mol Med.46:6 – 7 13.Fassati , P. and Prencipe , L. ( 1982 ) Measarement of serum triglycerides calorimetrically with an enzyme that p roduce H2O2 . Clin Chem 28 ( 10 ) : 2077 – 2080 . 14.Burstein M.; Scolink , H.R. and Morfin , R. : Measurement of HDLc in the plasma with a sensitive colorimetric method . J . Lipid Res. 1970 ; 19 : 583. 15.Friedwald ,W.T. : Kevy , R.L. and Fredrikson ,D.S. ( 1972 ) . Estimation of the concentration of LDL in plasma without use of preparative ultra centrifugation . Clin. Chem. ,18 : 499 – 502. 16.Hall , J.E. ; Taylor , A.E. and Hays F.J. ( 1998 ) .Insights into Hypothalamic – pituitary dysfunction in polycystic ovary syndrome . J. Endocr . Invest, 21:602 – 611. 17.Lobo , R.A. and Carmina , E, ( 2000 ) .The importance of diagnosing the polycystic ovary syndrome . Ann Intem Med 132 (12) : 989 – 993 . 18.Yen. S.C.C.( 1986 ) Chronic an ovulation due to CNS – Hypothalamic – pituitary dysfunction .Reproductive endocrinology ,physiology , path physiology and clinical management ; Jaffe , R.B. ( Eds. ) : P 500 – 545 . 19.Mchleilly , A.S. ( 1987 ) . Prolactin and the control of gonadotrophin secretion . J. Endocrinal , 115 : 1-5 . 20.Danielson ,K.K. ; Palta , M. ; Allen , C. ; and Dº Alessio , D.J. ( 2005 ) : " The Association of Increased Total Glycosylated Hemoglobin Levels with Delayed Age at Menarche in Young Women with Type 1 Diabetes " J. Clin Endocrinal .Metab., 90 (12) :6466 – 6471 . 21.Fiances , S.G. and David , G.G. ( 2001 ) . " Basic and Clinical Endocrinology " Disorder of lipoprotein , catabolic " ( 6 th Edition ) .( Chapter 20 ) :7230 . 22.Mather ,K.J. ; Kwan , F. and Corenbluw , B ( 2000 ) . Fertile , Sterile . 73 : 150 – 156 . 23 .Julie , L.Sh ( 2003 ) Clinical Diabetes , 21 ( 4 ) : 154 – 161. 24 .Pirwany , L.R. ; Fleming , R. , and Greer, I.A. ( 2001 ) . Clin.Endocr. ( Oxfo) , 54 : 447 – 453 ( Medline ) . 25.Weiss – M esser ,E.; Ber , R. and Barkey , R.J. ( 1996 ) .Endocrinology . 137 : 5509 – 5510 IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 Table ( 1 ) Mean± S .D‚ range and distribution of( LH , FSH and PRL) levels in serum of infertile female and healthy control Hormone LH ( IU/L ) Levels Number of subjects (%) of subject Mean ± S.D. Min. value Max value P- value Healthy Control 17 100 5.7± 1.76 0.7 7.8 Normal 5 25.6 6.7± 1.4 4.2 7.7 High 12 74.4 11.3± 2.6 8.4 17.5 Low Nil Nil Nil Nil Nil Patients Total 17 100 9.71±3.34 4.2 17.5 P<0.05 FSH ( IU/L ) Healthy Control 17 100 7.1±2.13 1.3 10 Normal 4 28 7.7 ±2.03 5.5 10 High 13 72 14.1± 3.1 10.2 21 Low Nil Nil Nil Nil Nil Patients Total 17 100 12.48±3.91 5.5 17.7 P<0.05 PRL (ngm/ml) Healthy Control 17 100 14.1± 3.05 7.03 18.2 Normal 5 33.4 12.6 ± 1 11 14 High 11 60 21.2± 3.8 15 29 Low 1 6.6 7± 0.00 7 7 Patients Total 17 100 17.56± 5.58 7 29 P<0.05 Table ( 2) BMI for infertile female and control group Groups No. Of subjects BMI ( Kg/m² ) ±S.D Patients 17 27.00 ±4.72 Control 17 25.8± 2.68 P-Value -------- P>0.05 - Each value represents mean ± standard deviation(S .D ) Table ( 3 ) Serum level of lipid profile of infertile females and control subjects Groups No. Of subjects Tc ( mg/dl ) ± S.D TG ( mg/dl ) ± S.D HDLc (mg/dl)± S.D LDLc ( mg/dl ) ± S.D VLDLc ( mg/dl ) ± S.D Patients 17 172.125± 35.58 108.68 ±31.63 57.75 ±7.56 97.68 ±27.14 21.68± 6.33 Control 17 171± 14.83 80 ±7.19 54.7± 7.39 100.1 ±10.38 16± 1.438 P-Value -------- P>0.05 P<0.05 P>0.05 P>0.05 P<0.05 Each value represents mean ± standard deviation(S .D ) IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 Fig(1):Distributions of (PRL, LH, &FSH)levels in serum of infertile female and healthy control Fig( 2):Distribution of infertile female according to( BMI ) Fig.(2):Distributions of infertile females according to (BMI) Fig.(3): Serum level of lipid profile of infertile females and control subjects 14.1 17.56 1.76 9.712 7.1 12.48 0 5 10 15 20 PRL LH FSH Control patient 6.70% 33.30% 40% 20% 0.00% 10.00% 20.00% 30.00% 40.00% % patients <18.5 18.5-24.9 25-29.9 >30 16 21.68 100.1 97.68 54.7 57.75 80 108.68 171 172.125 0 20 40 60 80 100 120 140 160 180 VLDL LDL HDL TG Tch control patients 2009) 4 (22مجلة ابن الهیثم للعلوم الصرفة والتطبیقیة المجلد ة الجسم مع الخلل في بعض هرمونات العالقة بین صورة الدهون ومؤشر كتل الغدة النخامیة في أمصال نساء عراقیات عقیمات عبد الرحمن رشید محمود جامعة بغداد، ابن الهیثم –كلیة التربیة ، قسم الكیمیاء الخالصة ة تـم تأكیـد التـداخالت بـین هرمونـات الهرمــون ،) LH(هرمونـات الجـسم االصـفر{ الجریبـات فـي هـذه الدراسـ أمــرأة عراقیــة عقــیم مــع صــورة الــدهون ، ) 17( فــي أمــصال })PRL(والهرمــون المحفــز للحلیــب ) FSH(المحفــز للجریبــة ــیطرة تــشمل ال عــن ضفــ ـة سـ ـارهن مقاربــة لعمــر النـــساء ) 17( مجموعـ أمــرأة بحالــة صـــحیة جیــدة وغیــر عقیمــات ، وأعمــ .العقیمات ــعاعي المنــاعي ـا صــورة الــدهون ) IRMA(أســتعملت تقنیــة التحلیــل االشـ ــات المــذكورة فــي أعاله،أمــ تـــشمل فلقیــاس الهرمونـ ـــي ــات الـــشحمیة العالیـــة، )TG(، الكلیــسریدات الثالثیـــة )Tc(مــستوى مـــصل الـــدم مـــن الكولیـــستیرول الكل الكثافـــة البروتینـ )HDLc (ـة ، وائــ الطرال باســتمت، فقــد قــدر اســتخدمت المعــادالت الریاضــیة فــي تقــدیرالبروتینات الـــشحمیة ق اللونیـ .قدر بطریقة حسابیة BMI(أن مؤشر كتلة الجسم ). VLDLc(والواطئة الكثافة جدا ) LDLc(واطئة الكثافة مــنهن %) 40( حـین أن فـي ,خلـل هرمـوني یعـانین الـسمنةلفقـط مـن النـساء العقیمـات نتیجـة %) 20(اظهـرت النتـائج ان .ات أوزان أعلى من الطبیعي وذ مـع اقـرانهن مـن عنـد النـساء العقیمـات بالمقارنـة)TG(أظهـرت لنـا الدراسـة ارتفاعامعنویـا فـي مـستوى الكلیـسریدات الثالثیـة .النساء الالتي بحالة صحیة جیدة ة فـي مــستویات الكولیــستیرول الكلــي الــشحمیة العالیــة البروتینــات ،)Tc(مـن ناحیــة أخــرى لــم تظهــر أي فــروق معنویــ .بین المجامیع قید الدراسة ) LDLc(والبروتینات الشحمیة واطئة الكثافة ) HDLc(الكثافة .أو السمنة التقترن مع العقم دائما) BMI(یمكن ان نستنتج من النتائج اعاله بأن مؤشر كتلة الجسم