Dichlorodiphenyltrichloroethane and the Adrenal Gland: From Toxicity to Endocrine Disruption

Endocrine disruptors are exogenous compounds that pollute the environment and have effects similar to hormones when inside the body. One of the most widespread endocrine disruptors in the wild is the pesticide dichlorodiphenyltrichloroethane (DDT). Toxic doses of DDT are known to cause cell atrophy and degeneration in the adrenal zona fasciculata and zona reticularis. Daily exposure in a developing organism to supposedly non-toxic doses of DDT have been found to impair the morphogenesis of both the cortex and the medulla of the adrenal glands, as well as disturbing the secretion of hormones in cortical and chromaffin cells. Comparison of high and very low levels of DDT exposure revealed drastic differences in the morphological and functional changes in the adrenal cortex. Moreover, the three adrenocortical zones have different levels of sensitivity to the disruptive actions of DDT. The zona glomerulosa and zona reticularis demonstrate sensitivity to both high and very low levels of DDT in prenatal and postnatal periods. In contrast, the zona fasciculata is less damaged by low (supposedly non-toxic) exposure to DDT and its metabolites but is affected by toxic levels of exposure; thus, DDT exerts both toxic and disruptive effects on the adrenal glands, and sensitivity to these two types of action varies in adrenocortical zones. Disruptive low-dose exposure leads to more severe affection of the adrenal function.

Effects of Estrogen and Exercise Stress on Adrenal Glands of Male Rats

Consciously done exercises are beneficial for human health. However, unconscious and wrong practices (medication use, etc…) can cause various injuries and permanent damage to the human body. For reasons such as increasing muscle mass and/or to getting more performance, taking steroid hormones disrupts overall body hormone balance. Effects of exercise on oxygen and energy metabolism and estrogen as an exogenous steroid have significant effects on the adrenal gland. The aim of this study was to see the effects of 17b-estradiol on surrenal glands of rats that is put through regular physical exercises. 36 male Sprague-Dawley rats between the weights 220-250 gr, were put into thermal-controlled room with day-night cycles to stimulate an optimal day for the subjects. The experiment was modeled by dividing 36 animals into 6 groups in total according to the control, exercising and estrogen administration criteria. Experimental group animals received daily doses of 10 μg/kg/day 17-β-estradiol during 30 days. Also exercises group animals ran at 20 m/min on a 15% grade for 90 minutes and rest 34 minutes. Then, TUNNEL and Hematoxylen & Eosin staining were performed to measure the damage on the adrenal glands. In group 2, dense presence of degenerative fibroblasts and inflammatuary cells infiltration in zona fasciculata were significantly different. In group 3, the degenerative areas were significant in all adrenal cortex zones. In group 4, necrotic areas were determined in zona reticularis. In group 5, zona fasciculata was severely degenerated. With group 6, Sinusoidal features were completely lost in zona reticularis. The results strongly show that exercise may affect the zona glomerulase in short time period. As a result, exposure to exercise and exercise stress with external administration of estradiol may cause cellular degeneration especially zona fasciculata and zona reticularis in the adrenal gland.

From adrenarche to aging of adrenal zona reticularis: precocious female adrenopause onset

Objective Adaptive changes in DHEA and sulfated-DHEA (DHEAS) production from adrenal zona reticularis (ZR) have been observed in normal and pathological conditions. Here we used three different cohorts to assess timing differences in DHEAS blood level changes and characterize the relationship between early blood DHEAS reduction and cell number changes in women ZR. Materials and methods DHEAS plasma samples (n = 463) were analyzed in 166 healthy prepubertal girls before pubarche (<9 years) and 324 serum samples from 268 adult females (31.9–83.8 years) without conditions affecting steroidogenesis. Guided by DHEAS blood levels reduction rate, we selected the age range for ZR cell counting using DHEA/DHEAS and phosphatase and tensin homolog (PTEN), tumor suppressor and cell stress marker, immunostaining, and hematoxylin stained nuclei of 14 post-mortem adrenal glands. Results We confirmed that overweight girls exhibited higher and earlier DHEAS levels and no difference was found compared with the average European and South American girls with a similar body mass index (BMI). Adrenopause onset threshold (AOT) defined as DHEAS blood levels <2040 nmol/L was identified in >35% of the females >40 years old and associated with significantly reduced ZR cell number (based on PTEN and hematoxylin signals). ZR cell loss may in part account for lower DHEA/DHEAS expression, but most cells remain alive with lower DHEA/DHEAS biosynthesis. Conclusion The timely relation between significant reduction of blood DHEAS levels and decreased ZR cell number at the beginning of the 40s suggests that adrenopause is an additional burden for a significant number of middle-aged women, and may become an emergent problem associated with further sex steroids reduction during the menopausal transition.

REGULATION OF ADRENAL RETICULARIS CELL’S PROLIFERATION IN ORGANISM DEVELOPMENTALLY EXPOSED TO LOW DOSES OF DDT

Altered patterns of cell proliferation in adrenal zona reticularis during postnatal development and regulation of proliferative activity by transcriptional factor PRH were revealed in rats exposed to endocrine disrupter DDT.

Biometry of Adrenal Glands of Mixed-Breed Dogs

Background: Differences of the size of the adrenal glands, in healthy dogs or in endocrinopathies, undermine correct diagnosis of endocrine disorders and evaluations of the adrenal cortex in relation to its size and possible correlation with endocrinopathies are rare. The aim of the present study was to perform measurements of the length, width, thickness and weight of the adrenal glands of young, adult and elderly mixed-breed dogs and correlate them with the age, sex and weight of animals. In addition, the areas occupied by the zona glomerulosa, zona fasciculata, and zona reticularis of the adrenal cortex were measured in order to establish a microscopic biometric pattern.Material, Methods & Results: The right and left adrenal glands of 12 young (six females and six males), 12 adults (six females and six males), and 12 elderly (six females and six males), all mongrels, derived from routine and necropsied in the Animal Pathology Sector of the Veterinary Hospital of the Federal University of Espírito Santo (HOVET-UFES) were weighed and the length, width, and thickness were measured. For the microscopic measurement of the adrenal cortex, 10 randomly selected samples were submitted to routine histological processing and the microscope slides were observed under a light microscope at 5× and 10× objectives, photodocumented and measurements were obtained from a random portion of the adrenal cortex and the zones composing the cortex were measured in triplicate with the aid of the computerized image analysis software. The left adrenal showed a greater average length than the right adrenal in young, adult, and elderly dogs. The size of the glands in the young and adult dogs is not influenced by the sex of the animals, but in older dogs the females had a greater mean width than the males. The weight of the animals presented a positive correlation in relation to the length and weight of the right and left adrenal glands in all studied groups. The age did not influence the length and weight variables of the glands. However, some differences in thickness and width were observed in the elderly group compared to those in other groups. For the microscopic measurements in the right adrenal gland, the cortex was 1.53 mm, being 0.21 mm for the glomerular zone (14.6% of the total adrenal cortex), 1.04 mm for the zona fasciculata (66.9%), and 0.29 mm for the zona reticularis (18.5%). In the left adrenal gland, the cortex was 1.83 mm, being 0.23 mm for the glomerular zone (13.2% of the total adrenal cortex), 1.23 mm for the zona fasciculata (63.96%), and 0.37 for the zona reticularis (22.84%).Discussion: Studies on the size of the adrenal gland in dogs using macroscopic biometrics are scarce, and the current study presents results regarding mixed-breed (male and female) dogs of different ages, which presented variation in size and weight, which could also influence the size of the adrenal gland. Thus, it can be observed that a variation of measurements can be found, especially for dogs with no defined breed. These results demonstrate the importance of studies that perform the macroscopic analysis of such glands. It was concluded that there was a clear variation in the size of the adrenal glands in mixed-breed dogs of different ages and both sexes; the left adrenal showed greater lengths and weights than the right gland. The sex of the animals did not influence the size of the glands in young and adult dogs, but in elderly dogs, the females showed a greater mean width than the males. The right and left adrenal cortices are formed at a greater percentage by the zona fasciculata, followed by the zona reticularis and zona glomerulosa.

MON-171 Characterization of the Adrenal Gland and Adrenal Rest Tissues in Congenital Adrenal Hyperplasia

Background: Adrenonodular hyperplasia and tumor formation are common long-term complications of congenital adrenal hyperplasia (CAH) driven by chronic ACTH elevation. Clinical studies indicate that the majority of males with classic CAH have testicular adrenal rest tumors (TART). Ovarian adrenal rest tumors (OART) are less commonly observed. Little is known about the pathophysiology of adrenal rest, however both adrenal cortex and Leydig cell markers have been described in TART, suggesting a pluripotent embryological cell origin. Objective: To characterize adrenals and adrenal rest tissues of patients with CAH in comparison with normal tissues. Materials and Methods: Using immunohistochemistry (IHC) and, real-time qRT-PCR we investigated CAH-affected adrenals (n=5), adrenal rest tissues (n=2; 1 testicular, 1 ovarian), controls [normal adrenal (n=2), testis (n=1), and ovary (n=1)]. Tissue sections prepared from paraffin embedded tissue blocks were immunostained with adrenal [melanocortin 2 receptor (MC2R), delta-like homolog 1 (DLK-1), steroidogenic factor 1 (SF-1), steroidogenic acute regulatory protein (StAR) and other cytochrome P450 genes], inflammatory [interleukin 2 receptor (IL-2R), B-lymphocyte antigen (CD20), cluster of differentiation 3 (CD3), tumor necrosis factor alpha (TNFα), interleukin 6 (IL6)], and gonadal markers [progesterone receptor (PR), androgen receptor (AR), insulin-like 3 (INSL3)]. RNA was isolated and gene expression studies were performed. High-throughput RNA sequencing technology was used to analyze the differential transcriptome profiles between the CAH adrenals, adrenal rest and normal tissues. Results: In contrast to the controls, CAH adrenals and adrenal rest tissues showed the following: (i) IHC studies revealed 95% of tissue positive for adrenal zona-reticularis; (ii) significant nodular lymphocytic infiltration with a predominance of B and T lymphocytes and overexpression of lymphocyte markers IL-2R, CD20, CD3, and inflammatory cytokines TNFα and IL6; (iii) increased expression of adrenocortical specific genes MC2R, DLK1. Conclusion: CAH-affected adrenals and adrenal rest tissue have similar predominance of zona reticularis and demonstrate lymphocytic infiltration. Active inflammation may play a role in the abnormal development of adrenal and adrenal rest tissue in CAH patients.

Morphofunctional rearrangements of adrenal cortex of adult rats at the experimental microelementosis

Xenobiotics, including salts of heavy metals, determine the development of pathology of individual organs and systems of the body. Adrenal hormones occupy a key place in the regulation and maintenance of the organism basic functions. The study of morphological, biochemical and immunohistochemical changes in the adrenal cortex of pubescent male rats at exposure to the complex of heavy metal salts remains an actual aspect of modern morphology. The experiment involved 24 white adult male rats weighing 250–300 g, aged 7–8 months. The animals of the experimental group consumed ordinary drinking water saturated with a combination of heavy metal salts (Zn, Cu, Fe, Mg, Cr) for 60 days. Histological, biochemical (determination of the content of hormones COR – cortisol, DHS – dehydroepiandrosterone sulfate) and immunohistochemical (determination of the expression of proliferation marker Ki-67) methods were used. Long-term consumption of the combination of heavy metals salts by mature rats leads to thickening of the stromal component of the gland, the rupture of capillaries and the violation of rheological properties of blood. Chromatin condensation, its marginal location, and initial stages of necrobiotic changes have been found in the spongiocyte nuclear apparatus. For endocrinocytes of the zona glomerulosa and zona reticularis of the gland, a slight increase in proliferative activity has been revealed, while cells of the zona fasciculata were non-reactive. These results correlate with the results of biochemical research, according to which hormones of the zona reticularis – DHS were actively involved in the formation of adaptive reactions in the body of experimental animals at the 60-day period of administration of the complex of heavy metals salts. The results of complex studies indicate a weakening of the secretory activity of cells of the zona fasciculata, which adversely affects the development of compensatory-adaptive processes in the body and the course of the general adaptation syndrome in response to the action of the damaging agent.