Effects of L-arginine on fatty diet induced changes in adrenal cortex: a morphometric study

To study the effects of L-Arginine on high fat diet induced changes in adrenal cortex. A prospective experimental study. This study was conducted at the Department of Anatomy, Basic Medical Sciences Institute, Jinnah Post Graduate Medical Centre Karachi from August 2008 to October 2008. Thirty male adult albino rats were taken for the study and were divided into three groups according to the dietary regimen. Group A received control diet. Group B received high fat diet with 20% added fat in the form of butter. Group C received high fat diet along with L-Arginine 300mg/kg/ day orally. After the end of the study period that is 8 weeks, animals were weighed and sacrificed. The adrenal glands were removed, fixed in buffered neutral formalin and after processing embedded in paraffin to form blocks. 4 micro m thick sections were cut and stained with H and E and Mallory's trichrome stains for morphometeric study. Highly significant increase in weight [P<0.001] was observed in animals of Group B and moderately significant [P<0.01] decrease was observed in Group c animals when compared to control and group B animals respectively. Cortical enlargement was found in zona glomerulosa and fasciculate and decreased thickness was observed in zona reticularis in group B animals, when compared to control, the results were highly significant [P<0.001]. In group C decreased cortical thickness was noted in zona glomerulosa and fasciculata but in zona Mallory's trichrome stained sections of Group B animals showed dilated blood vessels in the three cortical zones, more numerous in zona fasciculate when compared to control animals. In Group C no marked change was observed in all three cortical zones when compared to control. L-Arginine restricts the excessive weight gain caused by high fat diet. It also ameliorates the hypertrophic and vasodilatory effects on adrenal cortex caused by high fat diet

Ação da melatonina sobre a apoptose e fator de crescimento endotelial vascular no córtex da adrenal de ratas pinealectomizadas / Melatonin action in apoptosis and vascular endothelial growth factor in adrenal cortex of pinealectomized female rats

OBJETIVO: avaliar os efeitos da melatonina na apoptose e atividade celular no córtex da adrenal de ratas pinealectomizadas. MÉTODOS: foram utilizadas 40 ratas adultas, divididas randomicamente em 4 grupos com 10 animais cada: GCont - sem intervenção cirúrgica e com ministração do veículo; GSham - sem retirar a pineal com ministração do veículo; GExpV - pinealectomizado e tratado com veículo; GExpM - pinealectomizado e tratado com melatonina (10 µg/animal, por dia) durante a noite. Após 60 dias de tratamento, todos os animais foram submetidos à eutanásia, as adrenais retiradas, fixadas em formol a 10 por cento e processadas para inclusão em parafina. Nos cortes histológicos com 5 µm de espessura, foram realizados métodos imunoistoquímicos para detecção da apoptose (Caspase-3-clivada) e da atividade celular (fator de crescimento endotelial vascular, VEGF-A). Para avaliar a percentagem de apoptose foram contadas as células reativas em 1.000 células de cada zona da região cortical da adrenal em cada animal; já o VEGF-A foi expresso em escores. Os dados quantitativos foram analisados pela análise de variância (ANOVA) e pelo teste de comparações múltiplas de Tukey-Kramer (p<0,05). RESULTADOS: na avaliação do índice apoptótico, não foram encontradas diferenças significantes na zona glomerulosa entre os vários grupos estudados. Na zona fasciculada (GExpV=15,51±3,12*, p<0,05) e na reticulada (GExpV=8,11±1,90*, p<0,05) houve redução no índice apoptótico no grupo GExpV em relação aos demais grupos de estudo. Na reatividade ao VEGF-A, houve maior positividade, indicando maior atividade celular na zona na fasciculada do GExpV em relação aos outros grupos. CONCLUSÕES: a melatonina atua na região cortical da adrenal, em especial nas zonas fasciculada e reticulada, que estão relacionadas com a produção de esteroides sexuais.

PURPOSE: to evaluate the reactivity of VEGF-A and cleaved caspase-3 in the adrenal gland cortex of female pinealectomized rats treated with melatonin. METHODS: forty adult female rats were divided into 4 groups (G) of 10 animals: GI - no surgical intervention, with vehicle administration; GII - sham pinealectomized with vehicle administration; GIII - pinealectomized with vehicle administration; GIV - pinealectomized with melatonin administration (10 µg/animal) during the night. After 60 days of treatment, all animals were anesthetized, and the adrenal glands were removed and fixed in 10 percent formaldehyde (phosphate buffered) for histological processing and paraffin embedding. Sections (5 µm thick) were collected on silanized slides and submitted to imunnohistochemical methods for the detection of cleaved caspase-3 (apoptosis) and of vascular endothelial growth factor (VEGF-A) in the adrenal cortex. The data obtained were submitted to analysis of variance (ANOVA) complemented by the Tukey-Kramer test (p<0.05). RESULTS: reactivity to cleaved Caspase-3 was noted in the zona glomerulosa of the adrenal glands in all studied groups. There were no significant differences in the zona glomerulosa; however, the zona fasciculata (15.51±3.12*, p<0.05) and the zona reticularis (8.11±1.90*, p<0.05) presented the smallest percentage of apoptosis in the pinealectomized group (GIII). The reactivity to the VEGF-A was stronger in the zona glomerulosa and weaker in the zona reticularis in all groups. We found a stronger VEGF-A reactivity in the zona fasciculata in the pinealectomized group (GIII). CONCLUSIONS: the pineal gland affects the arrangement of the zona glomerulosa and reticularis of the adrenal glands, which are related to the production of sex hormones.

comparative study of the effect of Budesonide versus prednisolone on adrenal gland of the male rabbit

Although oral glucocorticoids are the treatment of choice for moderate to severe pancolitis, their systemic side effects and adrenal suppression account for considerable morbidity. Budesonide is a new intestinal topical active glucocorticoid which displays high therapeutic efficacy and high systemic tolerability. To deliver the active drug to ileal and ileocecal area, Budesonide has been formulated into enterocapsule preparation. Several studies compared the efficacy of Budesonide with that of Prednisolone. However, few determined the extent of adrenocortical suppression occurring with both drugs on histological basis. In this work, fifteen adult male New Zealand albino rabbits were used. They were classified into three equal groups. Group I served as control. Group II included animals that received intragastrically one tablet of 5 mg prednisolone daily for four weeks. Group III included animals that received orally one capsule of entocort containing 3 mg budesonide every other day for four weeks. The adrenals were processed for histological and immunohistochemical study. In the present comparative study, evidence of adrenal suppression was significantly greater in the prednisolone group than in budesonide treated animals. Light microscopic examination of H and E stained sections of prednisolone group, revealed an apparent decrease in the size of zona fasciulata cells which was proved significant by morphometric study. Moreover, Oil red stained sections of group II demonstrated a relative decrease in cytoplasmic lipid content of both zona glomerulosa and fasciculata. Using chromaffin reaction, it was noticed that there was a relative increase in the number of nor-epinephrine cells than in group III. Immunohistochemical study showed that most of the nuclei of cells in zona fasciculata in group II were negatively stained for proliferating cell nuclear antigen [PCNA] which was further proved significant morphometrically using the PCNA index. Thus it was concluded that, the budesonide preparation is associated with much less impairment of adrenal axis function. Therefore, budesonide offers a useful advance in treatment of inflammatory bowel disease

effect of Methylcobalamin on the adrenal medulla

Methylcobalamin is one form of cobalamin that acts as a nerve growth factor. Since adrenal medulla is part of the nervous system, therefore, this study was designed to assess the effect of methylcobalamin on this gland in rats. Albino rats were injected with methylcobalamin for 14 days; adrenals were taken, fixed in Ortho's fixative, and embedded in paraffin. They were cut in thin sections and stained with H and E and Giemsa stain. They were studied under light microscope. The result showed that methylcobalamin has stimulated the adrenal medulla, which was expressed by dilated vessels, and increase in the granularity and size of chromaffin norepinephrine containing cells. Sympathetic ganglionic cells have also increased in number and size. Cortical zona reticularis was hypervascular and hypertrophied. This study suggests that, methylcobalamin might have acted directly through the sympathetic nerves on the adrenal medulla or indirectly on the medulla through the adrenal cortex resulting in an increase in number of norepinephrine containing cells and the ganglionic sympathetic cells

Efecto de dosis altas de budesonida y función suprarrenal de niños asmáticos severos / High doses of inhaled budesonide and adrenal function in children with severe asthma

Background: The use of inhaled steroids is common in the treatment of bronchial asthma in children. Aim: To assess adrenocortical function in children with severe asthma receiving inhaled budesonide for six or more months. Material and methods: Children with severe asthma that required 800 µg/day or more of inhaled budesonide and that did not required systemic steroids for more than six days in the last four months to control their disease, were studied. Serum cortisol was measured one hour after administration of 0.25 mg of ACTH (Cosyntropin®) intravenously. The test was considered normal if post stimulation serum cortisol was over 18 µg/ml. Results: Twenty children (aged 5 to 14 years, 15 males), were studied. The stimulation test was normal in 17 children. Conclusions: Alterations in adrenal function are present in a small proportion of asthmatic children who require 800 µg/day of inhaled budesonide.

Age-related changes in the in vivo adrenocortical production of cAMP and corticosterone in response to exogenous ACTH in long-evans female rats

Ageing effects on the in vivo cyclic 3, 5'- adenosine monophosphate [cAMP] production by the adrenal cortex were studied in the rat. Materials and Methods: Eleven old [from 23 to 29 months] and 13 young [from 4 to 5 months], dexamethasone pre-treated Long-Evans female rats received 5.0 mu.i. [1-24] ACTH/100g b.w. by intravenous injection. The plasma concentration of corticosterone as well as the adrenal contents in cAMP and corticosterone were measured, by radioimmunoassay, just before and 45 min after the [1-24] ACTH injection. Results: The basal plasma corticosterone level and the adrenal contents in corticosterone and cAMP were low and no group difference was observed. The [1-24] ACTH injection causes significant increases in the plasma corticosterone level and the glandular contents in corticosterone and cAMP, which were lesser in the old animals than in the young ones; the differences aged/young were approximately -37%, -18% and -55% respectively. Conclusions: These results suggest that the reduced stero‹dogene response of the adrenal cortex in the old rat, to an acute ACTH administration is at least partly due to a decrease in the cellular production of the principal second messenger of this hormone, i.e. the cAMP

Protection of adrenocortical activity by dietary casein in ether anaesthetized rats.

Adrenal delta5-3beta-hydroxysteroid dehydrogenase (delta5-3beta-HSD) activity and serum corticosterone level were significantly higher in rats fed with 5% casein or 4% albumin diets after 1 hr of ether anaesthetic stress as compared to the controls, 5% casein and 20% casein (equivalent to 4% albumin) respectively. Ether anaesthesia to 20% casein fed rats caused no change in adrenal delta5-3beta-HSD activity and serum corticosterone level when compared with controls fed 20% casein diet. The results suggest that high milk protein diet may prevent acute stress effects by protecting adrenocortical activity. The present investigation opens up a new area of management of stress.

Prevention of adrenocortical hyperactivity by dietary casein in rats exposed to forced swimming stress.

Adrenal weight, adrenal hydroxysteroid dehydrogenase activity and serum corticosterone level were significantly higher in rats fed with 5% casein diet after 7 days of swimming stress (45 min/day) as compared to their controls. All the parameters were similar to their control levels in rats receiving 20% casein diet and exposed to swimming stress. The results suggest that casein can play an important role in preventing adrenocortical hyperactivity in swimming stressed rats.

Effect of nicotine on the morphology of adrenal cortex

The morphological effects of nicotine on the adrenal cortex have been observed in this study to get supportive evidence whether nicotine has some effect on the adrenal cortex; as regards the morphology. Animals were divided into three groups, which comprised group A control group, group B treated group and group C placebo group. Among these animals half of the animals from each group were sacrificed after 40 days and the remaining half after 60 days of treatment. Group A was control group, group B was given Nicotine Sigma 1 mg/kg body weight daily intra peritoneally for 50 days and B2 for 60 days. Group C and C 1, were given the same quantity of saline intra peritoneally daily for same period. On autopsy adrenals were removed and fixed in normal saline. 5 micron thick sections in paraffin and 10 micron thick frozen sections were studied for the cortical zones, cell size in each zone and among of lipid in cortical cells. It was observed that in treated group there was no change in B1 group but there was a marked hypertrophy of fasciculata cells with increased width of zona fasciculata. To conclude, the present study supports the view that nicotine has stimulant effect on the adrenal cortex and prolonged administration of the drug can induce morphological changes in fasciculata cells

Morning steroid profile in children with congenital adrenal hyperplasia under different hydrocortisone schedules.

We studied 13 children with 21-hydroxyalse deficiency to explore the immediate potential suppressive effect of hydrocortisone dose schedule on the adrenal cortex. They were given 20 mg/m2 daily in a controlled trial. After random administration of a greater dose in the morning (7 patients) or at night (6 patients), we measured plasma levels of 17-hydroxyprogesterone, testosterone, and androstenedione at times-24, 0, 2, 4, and 6h. Considerable fluctuation of the steroid levels, unrelated to the drug intake, was observed. There was no statistically significant differences between the "morning dose" and "night dose" groups for any steroid. We conclude that; (i) the greater night dose did not avoid the 17-hydroxyprogesterone morning peaks, and (ii) the variation in plasma steroid levels is so marked that a single morning sample is unreliable to reflect the degree of adrenal suppression.

Effect of adrenaline administration on the histological structure, lipid and ascorbic acid contents of mouse adrenal cortex

The adrenal glands were taken from 9 male mice after a single intraperitoneal injection of 0.03/100 g body weigh of adrenaline hydrochloride solution at different periods of time 0.5 hour, one hour and 2 hours [3 at a time]. The histological structure, lipid ascorbic acid contents have been studied. The results obtained in the present study showed that, the most obvious changes were seen as follows: Dilatation of cortical sinusoids, loss of subglomerulosal zone, and transformation of the fasciculate cells from the clear to dark type. Depletion of sudanophilic materials from zona reticularis and zona fasciculata started at 0.5 hour after injection with maximal depletion at 2 hours after injection. A single adrenaline injection caused a rapid fall in ascorbic acid content at 0.5 hour after injection. One hour after injection there was complete ascorbic acid depletion from zona glomerulosa and outer zona fasciculata

Effect of cortisone administration on the cholesterol content and phosphomonoesterases activities of mouse cortex

After a single IM injection of cortisone, the adrenal glands of 9 male mice were taken at different intervals of time 3 hours, 2 days and 3 days [3 at a time]. The cholesterol content and phosphomonoesterases activities have been studied. The results obtained in the present study showed that, 3 hours after cortisone injection, depletion of cholesterol content was observed in both zona fasciculata and zona reticularis. Subsequent accumulation of cholesterol in the zone fasciculata then occurred. Cortisone injection caused a decrease in phosphomonoesterases activities 3 hours after the injection, but these activities increased after 2 days from the injection

Effect of cortisone administration on the histological structure and lipid content of mouse adrenal cortex

After a single IM injection of cortisone, the adrenal glands of 9 male mice were taken at different periods of time [3 hours, 2 days and 3 days, 3 at a time]. The histological structure and lipid content have been studied. The results obtained in the present study showed marked narrowing of the adrenal cortex compared to the medulla. This narrowing involved only zona fasciculata and zona reticularis. Zona glomerulosa, however, showed relative broadening in width. Depletion of sudanophilic materials initially occurred followed by a marked increase 2 days after injection

Efeito da rifampicina sobre a reserva adrenocortical de cortisol em pacientes com tuberculose / Effect of rifampicin upon the adrenocortical reserve of cortisol in patients with tuberculosis

A tuberculose é causa conhecida de insuficiência adrenocortical primária. A administraçäo de rifampicina, parte do esquema tríplice empregado no tratamento da tuberculose, induz a síntese de enzimas hepáticas que aceleram o metabolismo do cortical, Uma vez que a rifampicina pode preciptar crise adrenal em tuberculose com funçäo adrenocortical comprometida, avaliamos possíveis alteraçöes na reserva de corticol em 33 pacientes com tuberculose, subdivididos em dois grupos: Grupo I - 17 pacientes (13M/4F, 21-45 anos) foram estados antes e, novamente, 30 dias após instituiçäo do esquema tríplice; Grupo 2 - 16 pacientes (13M/3F, 18-64 anos) foram estudados ao fim de seis meses de tratamento e, novamente 30 dias após sua interrupçäo. Como controles utilizamos 32 indivíduos normais, pareados para sexo e idade. Em cada estudo determinamos os níveis séricos de cortisol antes e 50 minutos após estímulo com ACTH (250µg i.v.). Nenhum paciente apresentou clínica de insuficência adrenal ou calcificaçöes nas radiográfias de abdome. Os valores do cortisol basal e pós ACTH (em µg/dl, X ñ DP) no Grupo 1 foram: 20,3 ñ 8,5 e 32,9 ñ 8,5 (antes) e 16,3 ñ 5,1 e 30,3 ñ 6,4 (após 30 dias de tratamento); no Grupo 2 foram: 19,1 ñ 7,3 e 35,4 ñ 12,1 (após seis meses de tratamento) e 18,4 ñ 4,6 e 31,7 ñ 8,2 (30 dias após a internaçäo). Nos controles normais obtivemos: 11,3 ñ 3,4 e 26,3 ñ 4,8, definindo limites críticos inferiores (LCL) de 5,7 e 18,4 para cortisol basal e pós-ACTH, respectivamente. Nenhum paciente apresentou níveis basais ou pós-ACTH de cortisol abaixo do LCI. A instituiçäo (Grupo 1) ou interrupçäo (Grupo 2 do tratamento com rifampicina näo modificou a resposta do cortisol ao estímulo com ACTH. Os dois grupos näo diferenciaram quanto aos níveis e repostas do cortisol. Assim, reduçäo da reserva de cortisol näo foi detectada em nenhum de 33 pacientes com tuberculose estudados prospectivamente, nos quais o tratamento breve ou prolongado com rifampicina näo resultou em alteraçöes significantes da resposta ao estímulo com ACTH

Effect of an LH-RH analog on cat testis and adrenal cortex function

The LH-RH analog-LH-RH-A (des-Gly10, [D-Trp6]-LH-RH ethylamide) was administerd in pharmacological doses (20 µg/Kg, sc) to adult male cats for 15 days and its effect on testis and adrenal function was determined. Dayly adminitration of the analog promoted a 3-fold increase in plasma testosterone levels after 7 days, indicating stimulatory effect of LH-RH-A (mean ñ SD for 6 treated cats, 1.88 ñ 0.35 vs 0.51 ñ 0.08ng/ml for 6 control cats). After 15 days the LH-RH-A-treated group exhibited a similar plasma testosterone concentration as the control group (mean ñ SD, 0.96 ñ 0.35 ng/ml vs 0.88 ñ 0.39 ng/ml, respectively), similar testicular and adrenal weights and no significant differences in the spermatogenic process. However, semiquantitative analysis of the zona fasciculata of the adrenals from the LH-RH-A treated group showed a significant accumulation of a substance not stained by hematoxylin-eosin or Schiff periodic acid (mean ñ SD of index of accumulation was 3.50 ñ 0.4 for treated cats vs 2.20 ñ 0.3 for control cats). The present results show that pharmacological doses of LH-RH-A have an effect on the adrenal cortex of cats without modifying spermatogenesis or plasma testosterone levels

Effect of lithium chloride on adrenocortical activity in male rats: evidence of dose and duration dependent response.

Changes in the adrenal weight, adrenal 5-ene-3 beta-hydroxysteroid dehydrogenase (5-ene-3 beta-HSD) activity and serum levels of corticosterone were observed in male wistar rats after the treatment of lithium chloride in the doses of 100, 200 and 400 micrograms/100 g of body weight/day for 7, 14 and 21 days. The experiments indicate that 200 and 400 micrograms/100 g.b.w. administered for 14 and 21 days caused a significant stimulation in the activities of adrenal 5-ene-3 beta-HSD along with elevation of adrenal weights and serum levels of corticosterone. 100 micrograms of lithium chloride was not able to modulate the adrenal activity. Moreover, plasma levels of lithium remain in therapeutic range in this experiment at the doses of 200 and 400 micrograms/100 g body weight. Therefore, our data suggest that lithium can alter the adrenal activity within its therapeutic range according to the duration of treatment.

Effect of ethinyl estradiol and norethisterone acetate on adrenal and some other tissues of the rat

Alkaline phosphatase activity [AP] of the adrenal cortex of rats was determine under the effect of ethinyl estradiol [EE] and/or norethisterone acetate [NA], the two components of the contraceptive pill Gyn-Anovlar [Schering A.G. Berlin]. LA poathological study was also carried out to examine the effects of EE and NA on other tissues, mainly the liver, lungs, spleen and ovaries. EE in a dose of 10 micro/rat/day for 2 weeks caused a significant increase in the weight of the adrenal but no significant increase in the AP/gm cortical tissue. The 25 and 50 micro doses for the same period caused a significant increase in both adrenal weight and AP. When treatment was prolonged to 6 weeks no effect on adrenal weight or APO was detected. The same findings obtained with NA in a dose of 7 mg/rat/day for 2 weeks. The 14 mg dose of NA for the same period caused a significant increase in adrenal weight but no effect on AP. The 21 mg dose caused a significant increase in adrenal weight accompanied by significant decrease in AP/gm cortical tissue. Treatment with NA for 6 weeks caused a rise in adrenal weight and AP with the 7 mg dose, then a decline in AP with the 14 mg dose, and a decline in both adrenal weight and AP with the 21 mg dose. As regards the effects of EE and NA on other tissues, EE was found to have a powerful stimulatory effect on the reticulo-endo-thelial system [RES] as well as toxic effects on the liver. NA did not produce such lesions except with the large doses and prolonged periods of treatment. In addition NA produced cholestasis in the liver associated with staining of the liver cells with bile. Combination of EE and NA in the form of gyn-anovlar produced more powerful stimulation of RES and decreased the toxic manifestations of either component. As regards the ovaries, administration of 50 ug EE for 6 weeks produced profound hyperplasia of the granulose cells of the Graafian follicles and inhibited ovulation, however, NA did not inhibit ovulation. With gyn-anovlar, the effect of EE on the ovaries seemed to predominate and ovulation was inhibited