AL-Anbar J. of Agri. Sci. , Vol.: 5 No. (2) , 2007 HISTOLOGICAL STUDY OF ADRENAL CORTEX DURING PREGNANCY AND LACTATION IN ALBINO RATS Naheda I. H. Al-Dulaymi Deptt. of Anatomy and Histology – College of Medicien / University of Al-Anbar Abstract This study was done on 45 animal (20 male and 25 femal) albino rats. 25 femal were divided in to five groups, each with five femal rats: Group I, Group II, at first & second 10th day of pregnancy and Group III, Group IV at first&second 10th day of lactation and Group V was acontrol group The observations of this study were documented as following: Increasing in thickness of cortical zones with accompaned hyperplasia and hypertrophy during pregnancy and continous at first 10th day of lactation. There was no significant differences in the weight of adrenal glands, inspite of increasing in the weight of left adrenal gland, while significant increase in the body weight was recorded during pregnancy in the studied groups of rats. The objective of study was designated to know the histological changes in the adrenal glands during pregnancy and lactation and the influence of pregnancy and lactation on body weight and adrenal gland weight in albino rats دراسث ىسٌجٌث لقشرت الغدت الكظرٌث خالل الحهل والرضاعث فى جرذاو االلةٌىو ) ( الجرذاو الةٌضاء ًىاهدت إةراهٌن حهادي الدلٌه كمٌث الطب / جاهعث االىةار الخالصث 04 امدراست أجرٍث عني 45 حٍواى وخجبرً ( جرذ أال مببيٍو ) بوعدل 20 ذلر و40 أيثي . قسوث أل : ٌأيثي أمٌ وجب وٍع شرحث فٌ لل وجووعت 4 حٍوايبث ولوب ٍن , اموجووعت األومي فٌ امٍوه امعبشر وى امحول , اموجووعت امثبيٍت فٌ امٍووه امعشورٍى ووى امحوول اموجووعت امثبمثت فٌ امٍوه امعبشر وى امرضبعت , اموجووعت امرابعت فٌ امٍوه امعشرٍى وى امرضبعت, اموجووعوت امخبوست حٍوايبث سٍطرة . ولبيث يجبئج امدراست اموثبجت , حصول زٍبدة فٌ سوك طبقبث امقشرة رافقهب زٍبدة فرط امجيسج وزٍبدة حجه امخالٍب خالل وراحل امحول واسجورث امزٍبدة حجي اموراحل األومي وى امرضبعت. لوب سجنث امدراست زٍبدة وعيوٍت فٌ وزى امجسه بٍيوب لبيث غٍر وعيوٍت فٌ وزى امغد د املظرٍت عني امرغه ووى وجوود . زٍبدة فٌ وزى امغدة املظرٍت امٍسرى INTRODUCTION Pregnancy was considered a normal physiological state which requires women functional preparation (1) Physiological and histological alteration develop in many systems during course of pregnancy and lactation. Some of these changes may effect health of mother and fetus (2) such as phyeochromocytoma which result from adrenal hormones imbelance (3). Hyper function of adrenal cortex is due to adrenal cortical tumor or hyperplasia which resulting in the clinical picture of cushingś syndrome(4). Several investigators have reported that there is arise in the concentration of cortisol in the maternal blood of animals during pregnancy (5). The adrenal cortex secreted three ageneral classes of steroid hormones, which grouped according to the dominant action as glucocorticoid hormone from zona fasciculata, meneralocorticoid from zona glomerulosa and aldosteron from zona reticularis (6,7). MATERIALS AND METHODS The experiment was done on 45(20 male and 25 femal) albino rat. The animals were kept in the animal house under controlled temperature and light. Tap water, and food in form of bellet were accessible to them. They were divided in to five groups, each with five femal rats. The group are : Group I and Group II were sacrified at 10th & 20th day of pregnancy, Group III and Group IV were sacrified at 10th &20th day of lactation. Group V was acontrol group. Animals were anesthetized by diethyl ether and weighted by using sensitive balance/ Metter AE 160(Switzer land). Methods used for determination the day of pregnancy are: Vaginal pluge, and vaginal smear which considered, the day of pluge as day zero of pregnancy (8,9). Animals were dissected. Adrenal glands were weighted and small blocks one cubic centimeter was used for routin histological studies. Tissue blocks were sectioned by microtome / Bright, 5030 microtome (England). At five microne, floated on water bath (45°c) and mounted on slids. Heamatoxylin and Eosin (H & E) and thickness of zones of adrenal cortex were calculated by slid champer. Data was analyzed using Statistical Package for Social Science(SPSS), F test (ANOVA) was used to compare among the groups (10). RESULTS The presente statistical study shows increasing in thickness of zones (ZG, ZF.ZR), (p<0.05) of both adrenal glands during pregnacy and continouse at first 10th day of lactation and regrssing during second 10th day of lactation (Table 1). It was also observed that adrenal zones more than right adrenal gland (Table 1). Table(1):- Distribution of thickness zones of the adrenal cortex in the studied groups of rats.(Mm) Pregnancy Pregnancy Lactation Lactation Control 1st 10th day 2nd 10th day 1st 10th day 2nd 10th day Mean±SD Mean±SD Mean±SD Mean±SD Mean±SD Left adrenal 46.6±12.1 57.7±14.4 48.4±15.5 71.04±24.3 51.06±20.2 ZG a* Left adrenal 455.2±60.7 517.2±180.2 466.4±116.5 488.5±92.7 415.1.±160.1 ZF b* Left adrenal 163.2±37.9 289.6±23.2 182.04±38.9 224.2±22.7 186.5±75.8 ZR c* Rightadrenal 50.9±6.3 52.7±8.3 51.06±6.07 68.8±26.5 53.3±16.5 ZG d* Right adrenal 426.2±23.01 501.2±174.1 408.4. ±64.4 490.6±105.7 455.1±113.5 ZF e* Right adrenal 142.1±24.1 261.9±29.9 142.1±21.4 162.1±55.3 264.3±138.8 ZR f* A* there was no significant defferences in the studed groups P>0.05 F* there was significant defferences in the studed groups P<0.05 Microscopically: the study reveled that there is a histological changes in the adrenal cortex as seen in figures . ZG ZG ZF ZF ZR ZR Fig. (1):- Section of control animal shows normal Fig. (2):- Section shows hyperplasia with increase thick ness of appearance of ‘adrenal gland zones:ZG(zona glomerulosa ↓) ZG( ↓), ZF(↕) , ZR( ↑) during first 10 th day of pregnancy , ZF ( zona fasiculata ↖ ) ZF(zona fascicu ZR(zona reticularis ↓) H&E(X100) H&E(X100) ZG ZF ZF Fig. (4):- High power picture , show clear hyperplasia with Fig.(3):- Hyperplasia with hypertrophy in zona hypertrophy in zona glomerulosacells (ZG↔ ), and fasciculate cells ZF(↕)and sinusoidal dilatation hypertrophy in zona fasciculata cells(ZF→) during first 10th between the columns(→ ) , at second 10th day of day of lactation H&E(X400) pregnancy. H&E(X100) of lac ZG ZF ZR Fig. (5):- Section show regression in thickness and hyperplasia of cortical cells (ZG,ZF,ZR) to reach as control animals at second 10th dayof lactation. H&E(X100) Table 2: -Distribution of the body weight and adrenal glands weight in rats. Body weight Weight of adrenal (gm) glands(gm) Groups Right (b) left(c) (a) Mean±SD Mean ±SD Mean ±SD Virgin Control 185.3±55.13 0.017±0.002 0.018±0.002 Pregnancy 1st 10th day 236.3±34 0.018±0.007 0.090±0.157 2nd 10th day 250.2±14.7 0.020±0.008 0.015±0.007 1st 10th day 223.9±16.8 0.034±0.031 0.032±0.033 Lactation 2nd 10th day 194.4±20.5 0.012±0.002 0.022±0.003 Table 2:- showed significant differences (P<0.05) in the body weight,(a) while no significant differences(P>0.05) in the wight of adrenal glands(b,c) although there was an increase in the weight of left adrenal gland during pregnancy. . 260 240 220 Mean of body weight 200 180 first 10 days pregna fist 10 lactation cotrol second 10 days pregn 2nd 10 days lact FACTOR Figur (6):- Distribution of body weight in the studied. groups of rats. DISCUSSION Many changes occures during normal pregnancy in adrenal steroids which presented by increased anabolic and decrease catabolic of adrenal steroids, increasing the placental steroids (11). Other changes that occur during pregnancy, increasing in ACTH secretion that lead to stimulate corticosteroids secretion from zona fasciculata (5). This study showed an increasing in thickness of zones(Zona glomerulosa, fasciculata and reticularis), of adrenal gland cortex during pregnancy (Table 1). Our suggestion that this increasing in thickness is due to hormonal secretion from these zones. It is agreed with the finding of Fleischman(12) , Foreier(13), Brouchu (14), who suggested that the increasing plasma renin activity (P.R.A) stimulat zona glomerulosa to increase aldosteron secretion in plasma of pregnant rats. Also several investigators reported that there is aris in concentration of corticosteroid in pregnant rats and mouse (15,16,17). There was inverse relastion ship between secretion of cortisol and advance pregnancy(18),Gartner(19), and Hadden(3), reported that the zona reticularis was increased in secretion of steroid hormones such as estrogene and progesterone during pregnancy. Also our study provide continous an increasing of thickness of zones at first 10th day of lactation(Table 1). Functional adrenal cortex producing the hormone II oxygenated steroid which promote glucogeneogensis is to be essential for lactogenesis(20). In lactating mice the fasciculata cells show amarked increase in vaculation and enlargement in size(21) . Our findings provide morphological evidence as hyperplasia and hypertrophy that associated with increasing of thickness zones during pregnancy(Fig.2 ,fig3. ) which continued at first 10th day of lactation(Fig.4) , and diminished during second 10th day of lactation to reach as control (Fig. 5) . It was observed in hamster by Nowak(22), that total number of adrenal cortex cells was increased in 10th days of pregnancy and decreased after pregnancy. This study was in agreement with Kissane (23), Gruhm (24) ,who claimed that hyperplasia showed an increase in cortical cells . Hyperplasia of adrenal cortex is due to clinical picture of cushing syndrom (4). The mean concentration of prolactin increased during the 6-day periods after delivery, this might be due to increase in dexamethasone concentration (25). Our data demonstrated there was no significant differences in the weight of both adrenal glands(Table.2 ). A marked increase in weight of left adrenal gland during first 10th day of pregnancy and decline at second 10th day of lactation (Table 2). These changes were due to the morphological and callculated changes in adrenal cortex as hyperplasia and change in size. Our finding was in agreement with finding of Gruhm(24) , who demonstrrated the adrenal cortex hyperplasia lead to increase in cortical cells that usually lead to increase in weight and size of glands. Blood worth (26), Sloper(27), Symington(28), and Page(29) ,were showed that adrenal hyperplasia lead to increase weight of glands 5% of normal weight. The suggestion of the some of investigators were supporte the our study, that founde inverse realation shipe between the weight of adenal gland and thickness of zone The body weight during pregnancy and lactation Results obtained from the present study showed significant (P<0.05) Increase in the body weight (Table 2, Fig.6). It was suggested that the increasing in the hormonal activity during pregnancy, such as progesteron leads to weight gain (30). Its well known in the laboratory animals, such as rats, that the number of fetuses increased during pregnancy. This may lead to increase of the body weight of mother (31, 32,33). 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