IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 Association Between Lipid Profile ,BMI , and Some Pituitary Ho rmones Abnormalities In Sera of Iraqi Infertile Females A. R.Mahmood Departme nt of Chemistry, College of Education Ibn – Al-Haitham, Unive rsity of Baghdad Abstract In this st udy , the clinical imp act of interaction between gonadotrop hin hormones (luteinzing hormone, LH, and follicle st imulating hormone ,FSH ) and p rolactin PRL in serum of seventeen Iraqi infertile female with the lip id p rofile . In addition to control group involving age matched fertile females . Immunoradiometric assay ( IRM A ) technique for the determination of (LH , FSH and PRL) was utilized. The lip id p rofile { i.e. total cholesterol ( Tc ) , triglycerides (TG) , and high density lip op rotein – Cholest rol ( HDLc )} ,were evaluated by using colorimetric method , while{ low density lip op rotein cholesterol ( LDLc ) and very low density lip op rotein – cholesterol ( VLDLc )} , were evaluated by using a mathematical formulas. The body mass index ( BM I ) 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 group s under consideration . A conclusion could be obtained from the above data , that (BM I )or obesity is not associated with infertility alway s. Introduction The reproductive sy st em of women shows regular cyclic changes that may be regarded as p eriodic p reparations for fertilization and pregnancy . This cycle is t he menstrual cycle [1] . M enstruation is dependent on the p rop er functioning of a chain made up of ; hy p othalamus → p ituitary →ovary(HPO) ; amenorrhea p resup p oses awakening or break in one or more of these links [2] . Interaction ( HPO ) to p roduce cyclic exp ression of the app rop riate hormones at the exp ected time or chronology age at which sexual maturation should normally occur [3] The growt h and the reproductive activities of the gonadal tissue are controlled by the gonadotrop hins hormone LH and FSH from the interior p ituitary gland . LH and FSH are called gonadotrop hins because they regulate the function of the gonads (ovaries and test es ) , in both sexes , FSH st imulates gamete ( sp erm or egg ) p roduction, while LH p romotes p roduction of gonadal hormones [4] .The secretion of both LH and FSH is st imulated by gonadotrop hin releasing hormone from the hy p othalamus , LH and FSH bot h are subjected to feed back loops regulation by the ovarian hormones [5]. In women , LH along with FSH are ordered as p art of t he workup of infertility and also useful in the invest igation of menst rual irregularities , and to aid in the diagnosis delay ed and p recocious p uberty [6]. Prolactin PRL st imulates mammary development and subsequent lactation , it is an episodic secretion that it p roduced by the lactotrop hs of interior p ituitary[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 p resence of a disease , this would increase in PRL level in the blood above the p hy siological 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 hy p erandrogenism , although the mechanisms underly ing this relationship remain largely undetermined [ 9 ]. The aim of the p resent st udy is to evaluate the lip id p rofile of infertile woman with p ituitary hormones dy sfunction and compared with that for healthy fertile woman matched in age and body mass index BM I. Experimental Part Seventeen infertile female were selected from Elwy ia M aternity Teaching Hosp ital during 2008, a careful history was obtained from the p atients including menst rual disturbance , if any , associated with the sy mptom of infertility , weight and height and all p atients, were free from medication affecting hormone level . Evaluation of each p atient is done by detecting body mass index, serum gonadotrop hens hormones level LH , FSH and serum PRL. Excluding the test during the days (12 – 16)from the cy cle period . As control (17) normal female with normal regular menst rual 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 p lain tubes , centrifuged at 3000 rp m for 10 min, after allowing the blood to clot at room temp erature . The sera were a liquated and frozen until assay p erformed . Body mass index uses a mathematical formula based on p erson's height and weight , BM I equals weight in kilograms divided by height in square meters ( BM I = kg / m²) [10]. Fost et al suggested that a BM I of ( 18.5 – 24.9 ) indicates a person of normal weight . A p erson with a BM I of ( 25 – 29.9 ) is overweight , while a p erson with a(BM I of ≥ 30) is obese [11] . Immunoradiometric assay IRM A for the in vitro determination of FSH , LH , and PRL in human serum was utilized by kits from immunotech abeckman coulter comp any . The Immunoradiometric assay of luteinizing hormone FSH , LH , and PRL is a sandwich ty p e assay the kit utilizes mouse monoclonal antibodies directed against two different ep itop es of FSH , LH and PRL hence not comp eting . Samp les 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 asp irated and rinsed after incubation and bound radioactivity measured values are calculated by interp olation from the st andard curve . The radioactivity bound is directly p rop ortional to t he concentration of FSH , LH , and PRL in the samp les. The results are obtained from the st andard curve by interp olation. The st andard curve serves for the determination of FSH , LH and PRL concentration in samp les measured at t he same time as t he calibrator . Determination of total serum cholesterol [12] involves the use of three enzy mes , cholesterol esterase , cholesterol oxidase and p eroxidase . In the p resence of the former mixture, ( N- ethy l p rop y l – m – anisidine ) and 4 – amino – antip y rine are condensed by hy drogen p eroxide to form quinoneimine dy e prop ortional to t he concentration of cholesterol , when the absorbance of the samp les 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 enzy matic hy drolysis with lipases . The indicator is aquinoneimine formed from hy drogen p eroxide , 4-aminophenazone , and 4 – chlorop henol under the cataly tic influence of perioxidase . The absorbance was measured for test and st andard against the reagent blank within 60 minutes at 500 nm [13]. In determination of high density lip op rotein – cholesterol HDLc [14], the method uses a selective p recipitations of chylomicrones and the apolip op rotein containing lip op rotein VLDLc and LDLc by addition of 4% p hosp hotungst ic acid solution , which contain 10 % magnesium chloride PH 6.2 . Sedimentation of the p recipitant by centrifugation , and subsequent enzy matic analysis of HDLc as residual cholesterol remaining in the clear sup ernatant , from which the cholesterol can be determined as described as above according to [12]. Low density lip op rotein cholesterol LDLc was determined by using emp irical Friedwald formula which was based on the assump tion that VLDLc is p resent in serum at concentration equal to one fifth of t he TG concentration . This formula is as follow [15] :when all concentration are given in milligrams p er deciliter :- LDLc(mg / dl) = T otal cholesterol - ( HDLc +VLDLc ) S tatisti cal Analysi s of Data To comp are the significance of the differences in the mean values of any two group s st udent's t-test was app lied and P value less than 0.05( p <0.05) was considered statist ically significant . Re sults and Discussions Table (1)and fig(1) showed the mean distribution of serum hormones concentration LH , FSH , and PRL of the infertile female p atients with the minimum and maximum values to each hormonal level when comp ared with the mean concentration of the normal control . A significant difference was observed between normal and high hormonal levels of infertile female comp ared to the majority of infertile have hormonal levels higher than that of control with a p ercentage of 74.4%, on the other hand there was no significant differences in the level of FSH in 28% of infertile female comp ared with healthy control . The significant difference was found in the higher levels of FSH in 72% of infertile female comp ared with normal hormonal level of healthy control . The results agreed p artly with other studies who concluded that different results could reflect variation in the selection of p atients and / or the different lifesty le factors [16]. The mean serum level of LH / FSH ratio for p atients was(0.8 )and for control group was(0.8). The difference is not significant y et ot hers st udies rep orted higher or lower ratio and they st ated that this variation might be due either to p rimary central disorders involving (GnRH) secretion or secondary p ituitary sensitization to (GnRH) by an abnormal feed back signals from ovaries [17] . The PRL level in (60%) of infertile female showed a significant increase comp ared to that of healthy control while lower significant values than control was depicted in the rest of the p atient .High p rolactin level are found in( 30% )of women with different kind of amenorrhea leading to infertility [18] . About ( 60% )of p atients of high level of p rolactin showed lower levels of FSH and LH than healthy control. These results are in agreement with other st udies who suggested that a decline in gonadotrop hine in hy p erproctanemic p atient showed the association between gonadatrop hine deficiency and hy p erprolactinemia which might be an indirect sign of functional hy p othalamic p ituitary interrup tion 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 p rovider. IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (4) 2009 Table (2)represent mean±SD of BM I for both group s .Fig (2) showed the distribution of infertile female according to BM I.There was a significant difference between p atients, p atients were overweight (40 % ), and( 33.3 %) of p atients were in a healthy weight range ,while the p ercentage of obese p atient was( 20%). The p resent results disagree with some reports of infertility association with obesity [20 ] . Table (3) and fig (3) showed the lip id p rofile 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 resp ectively . The HDLc and LDLc were (57.75 ±7.56 mg / dl) and (97.68 ±27.14 mg / dl) in serum of the infertile female resp ectively , and were (54.7± 7.39mg/dl) and (100.1 ±10.38mg /dl )for the control group resp ectively . The values for VLDLc were (21.68±6.33mg/dl)and(16±1.438mg/dl) for the infertile females and control resp ectively .A non significant elevation in total cholesterol as comp ared with control group was found .The result does not agree with other st udies which found that women with different causes of infertility had increase Tc levels[21]. A significant elevation in TG in serum of infertility female p atient comp ared to control was obvious as shown in table( 3 ). The p att ern of dy slipidemia is mostly found in a wide range of causes of infertility , even st udies 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 consist ently 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 p atients with hy p erprolactinemia were reported due to acceleration of hepatic and adip ose tissue biosy nthesis of triglyceride and certain p hosp holip ids factors [ 25 ]. Re ferences 1.Ganong, W.F. ( 1999 ) .The Gonads Develop ment and Function of the Rep roductive Sy st em In : A review of M edical Phy siology ( 19 th edition) . Ap p leton and Lang M edical Publishment – Stamford , Connecticut .P 393 – 432. 2.Tindall ,V.R. ( 1987 ) . Amenorrhoe : Scanty and Infrequent menstruation . In : Jeffcoate's p rinciples of Gy naecology , ( 5 th edition ) . Butt erworth & Co. ( Publishers ) Ltd. P.495 – 496 . 3.James , D. and Brian , A.M . ( 2004 ) " Amenorrhea " .In " Clinical obst etrics and Gy naecology " , ( 1 st edition ) . Saunders . P 127 . 4.Elaine , N. and M arieb , R.N. ( 2000 ) : The Endocrine Sy st em in Anatomy and phy siology . Benjamin Cummings , San Francisco , Chapter 15: 512 , 518 . 5.Zilva , F. ; M ayne , D. Philip and Pannel ( 1999 ) The hy p othalamus and p ituitary gland .In : Clinical chemist ry in Diagnosis and Treatment , 6 th ed. Hodder Headline group PLC London , chapter 5: 106 – 115. 6.M c Donough ,P.G. ( 2003 ) : M olecular abnormalities of FSH and LH action , Ann N Y Acad Sci 997 : 22 – 34. 7.M arc , E.F.; Bela , k. ; Anna ,L. and Gy orgy , N. ( 2000 ) , p rolactin : Structure , Function and regulation of section : Phy siological Reviews 80 ( 4 ) : 1523 – 1631 8.Hanan , L.S. ( 2002 ) . The effect of hy p erprolactinema on Follicular development and estradiol concentration in stimulated cy cle .M Sc. Thesis , college of medicine . Univ. of Baghdad , Iraq .P.44 – 46 . 9.Franks , S. ; Kiddy , D. and Sharp , P. ( 1991 ) obesity and polycyst ic ovary sy ndrome . 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 p rinciples of Int ernal M edicine , Volume 1, ( 16 th edition ) . M cGraw Hill ,M edical 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 develop ment of an enzy matic technique for the assay of cholesterol in biological fluids .Clin Scc M ol M ed.46:6 – 7 13.Fassati , P. and Prencip e , L. ( 1982 ) M easarement of serum triglycerides calorimetrically with an enzy me that p roduce H2O2 . Clin Chem 28 ( 10 ) : 2077 – 2080 . 14.Burst ein M .; Scolink , H.R. and M orfin , R. : M easurement of HDLc in the p lasma with a sensitive colorimetric method . J . Lip id Res. 1970 ; 19 : 583. 15.Friedwald ,W.T. : Kevy , R.L. and Fredrikson ,D.S. ( 1972 ) . Est imation of the concentration of LDL in plasma without use of p reparative ultra centrifugation . Clin. Chem. ,18 : 499 – 502. 16.Hall , J.E. ; Tay lor , A.E. and Hay s F.J. ( 1998 ) .Insights into Hy p othalamic – p ituitary dy sfunction in p olycyst ic ovary sy ndrome . J. Endocr . Invest , 21:602 – 611. 17.Lobo , R.A. and Carmina , E, ( 2000 ) .The importance of diagnosing the polycyst ic ovary sy ndrome . Ann Int em M ed 132 (12) : 989 – 993 . 18.Yen. S.C.C.( 1986 ) Chronic an ovulation due to CNS – Hy p othalamic – p ituitary dy sfunction .Rep roductive endocrinology ,p hy siology , p ath p hy siology and clinical management ; Jaffe , R.B. ( Eds. ) : P 500 – 545 . 19.M chleilly , A.S. ( 1987 ) . Prolactin and the control of gonadotrop hin secretion . J. Endocrinal , 115 : 1-5 . 20.Danielson ,K.K. ; Palta , M . ; Allen , C. ; and Dº Alessio , D.J. ( 2005 ) : " The Association of Increased Tot al Gly cosy lated Hemoglobin Levels with Delayed Age at M enarche in Young Women with T y p e 1 Diabetes " J. Clin Endocrinal .M etab., 90 (12) :6466 – 6471 . 21.Fiances , S.G. and David , G.G. ( 2001 ) . " Basic and Clinical Endocrinology " Disorder of lip op rotein , catabolic " ( 6 th Edition ) .( Chap ter 20 ) :7230 . 22.M ather ,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 ( M edline ) . 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 distributi on of( LH , FS H and PRL) levels i n serum of infe rtil e female and healthy control Hormone LH ( IU/L ) Levels Number of subjects (%) of subject M ean ± S.D. M in. value M ax 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 infertil e female and control group Group s No. Of subjects BM I ( 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 ) S erum level of lipid profil e of infe rtil e females and control subjects Group s 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, &FS H)levels in se rum of infertile fe male and he althy control Fig( 2):Distribution of infertile female according to( BM I ) Fig.(2):Distributi ons of infe rtil e females according to (BMI) Fig.(3): S erum level of lipid profile of infertile females and control subjects 14.1 17.56 1.76 9.712 7.1 1 2.48 0 5 10 15 20 PRL LH FSH Con trol p atient 6.7 0% 3 3.30% 40% 2 0% 0.0 0% 10.00 % 20.00 % 30.00 % 40.00 % % patients <18.5 18.5-2 4.9 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 patie nts 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 الهرمــون ،) LH(هرمونـات الجـسم االصـفر{ الجریبـات فـي هـذه الدراسـة تـم تأكیـد التـداخالت بـین هرمونـات أمــرأة عراقیــة عقــیم مــع صــورة الــدهون ، ) 17( فــي أمــصال })PRL(والهرمــون المحفــز للحلیــب ) FSH(المحفــز للجریبــة أمــرأة بحالــة صـــحیة جیــدة وغیــر عقیمــات ، وأعمـــارهن مقاربــة لعمــر النـــساء ) 17( مجموعــة ســـیطرة تــشمل ال عــن ضفــ .العقیمات IRM(أســتعملت تقنیــة التحلیــل االشـــعاعي المنــاعي A ( تـــشمل فلقیــاس الهرمونـــات المــذكورة فــي أعاله،أمـــا صــورة الــدهون الكثافـــة البروتینـــات الـــشحمیة العالیـــة، )TG(، الكلیــسریدات الثالثیـــة )Tc(مــستوى مـــصل الـــدم مـــن الكولیـــستیرول الكلـــي )HDLc (اســتخدمت المعــادالت الریاضــیة فــي تقــدیرالبروتینات الـــشحمیة ق اللونیــة ، وائــ الطرال باســتمت، فقــد قــدر BM(أن مؤشر كتلة الجسم ). VLDLc(والواطئة الكثافة جدا ) LDLc(واطئة الكثافة I قدر بطریقة حسابیة. 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 الــشحمیة العالیــة البروتینــات ،)Tc(مـن ناحیــة أخــرى لــم تظهــر أي فــروق معنویــة فـي مــستویات الكولیــستیرول الكلــي .بین المجامیع قید الدراسة ) LDLc(والبروتینات الشحمیة واطئة الكثافة ) HDLc(الكثافة BM(یمكن ان نستنتج من النتائج اعاله بأن مؤشر كتلة الجسم I (أو السمنة التقترن مع العقم دائما.