Chronic unpredictable mild stress increases serum aldosterone without affecting corticosterone levels and induces hepatic steatosis and renal injury in young adult male rats.

Stress is often associated with anxiety and depressive symptoms in adolescents. Stress is associated with components of metabolic syndrome and inflammation. The present study hypothesizes that aldosterone, more than corticosterone, promotes chronic stress-hepatic steatosis and fibrosis, as well as renal inflammation and fibrosis in young adult rats. Thirty-two young adult male Wistar rats of 51 days old were divided into four groups (n = 8 per group): Control (C), chronic unpredictable mild stress (CUMS), control plus vehicle (C plus veh), CUMS plus eplerenone, a selective aldosterone blocker (CUMS plus EP). On postnatal day 51, eplerenone was administered orally through a gastric tube two hours before the start of the stress test. The CUMS paradigm was administered once daily at different times, with no repetition of the stressor sequence for four weeks. Renal inflammation and fibrosis were measured, as well as liver glycogen, triacylglycerol, and fibrosis levels. The serum concentrations of corticosterone, aldosterone, sodium, and creatinine were measured in urine and serum. The CUMS group showed a high level of serum aldosterone without affecting the level of corticosterone, increased urinary sodium, tubular atrophy, glomerular sclerosis, the presence of inflammation, and fibrosis, without affecting creatinine, increased glycogen content, triacylglycerol, and moderate fibrosis in the liver, and treatment with eplerenone prevented the inflammation, fibrosis, glycogen, and triacylglycerol. Our results show that chronic stress-induced aldosterone promotes hepatic steatosis and renal injury more than corticosterone. The prevention by eplerenone supports our hypothesis.

Sucrose consumption modifies the urethrogenital reflex and histological organization of the bulbospongiosus muscle in the male rat.

Disorders of the bulbospongiosus muscle (Bsm) are associated with male sexual dysfunction, such as premature ejaculation. We determined the effect of sucrose-water consumption during pregnancy-lactation and postnatal on reflex responses and morphology of Bsm fibers in adult male Wistar rat offspring. Female rats were mated and grouped into consumed tap water mothers and sucrose-water (5 %) mothers during pregnancy-lactation to obtain experimental groups. Male pups were weaned and assigned into four groups (n = 12; each group). Those from control mothers who continued drinking tap water (CM-CO group) or sucrose water (CM-SO group), and those from sucrose mothers who drank tap water (SM-CO group) or continued drinking sucrose water (SM-SO group) until adult life. In male rat offspring (n = 6 per group) was recorded the electrical activity of Bsm was recorded during penile stimulation and urethrogenital reflex (UGR). Other male rat offspring were designated for histological analysis (n = 6 per group). Sucrose consumption during prenatal stages increased the frequency of the Bsm during UGR, while pre and postnatal consumption modified muscle fiber cross-sectional area and increased the collagen content, suggesting that a combination of a diet with pre- and postnatal sucrose changes the Bsm morphophysiology possibly causing male sexual dysfunctions.

Dietary intervention in adult rats exposed to a high-sugar diet early in life permanently impairs sperm quality.

Childhood obesity predicts the presence of adult obesity. Obesity is associated with poor sperm quality. We hypothesized that exposure to a high-sugar diet (HSD) in early life would cause permanent histomorphology damage to the testes, resulting in reduced sperm quality in adult life. Wistar rats (aged 21days) were divided into four groups (n=6). In the first experiment, the rats received tap water (control) and a 30% sucrose diet for two months (S30). In the second experiment, the control and 30% sucrose diets were fed for two months, followed by replacement with tap water for two months (IS30). Eating and drinking were monitored. Animals were then euthanized, visceral and gonadal fat tissue and testes were collected, and epididymal spermatozoa were excised. Testicular samples were used for morphological description by H&E staining and for quantifying triacylglycerol content, caspase activity, and oxidative stress. Serum testosterone concentration was evaluated. Spermatozoa were used to assess sperm quality. Our results show that sperm quality was impaired by consuming HSD and could not be restored by dietary intervention. HSD feeding induced hyperplasia of visceral adipose tissue, increased testicular weight, and serum testosterone levels. The dietary intervention increased visceral adipose tissue, serum, and testicular triacylglycerol levels and normalized serum testosterone levels. Overall, the HSD diet caused permanent changes in seminiferous tubule cross-sectional area, caspase activity, oxidative stress, and sperm quality. Therefore, a high-sugar diet in early life causes permanent damage to sperm quality in adulthood.

High-sucrose diet-fed male rats show affectations in pubococcygeus reflex activation and myofiber content.

In males, the function of the pubococcygeus muscle (Pcm) shows a high androgen dependency. High sucrose intake increases testosterone levels in pubertal male rats. We aimed to analyze the Pcm reflex activity and fiber type in response to a 30 % sucrose diet. Wistar male rats were assigned into two groups: control and sugar. The Pcm electromyographic activity was recorded during the mechanical stimulation of the scrotal skin and penis. We also determined the relative levels of Glut4 expression, glycogen content, myofiber cross-sectional area (CSA), and the content of glycolytic and oxidative with NADH-TR or fast, intermediate, and slow fibers with alkaline (pH 9.4) ATPase histochemistry. The sugar group showed a short Pcm reflex activity, a higher Glut4 expression, and glycogen content; the CSA fibers showed a significant difference in the percentage of fibers with the different transversal areas. The number of nuclei was positively correlated with the CSA of the Pcm fibers. There was also a decrease in oxidative fibers but an increase in glycolytic fibers, while the content of fast muscle fibers increased to the detriment of intermediate and slow fibers. Thus, a high-sugar diet reduced Pcm reflex activity, fiber type, and dysregulation of skeletal muscle energy metabolism.

The Effect of Microbiome Therapies on Waist Circumference, a Measure of Central Obesity, in Patients with Type 2 Diabetes: A Systematic Review and Meta-analysis of Randomized Controlled Trials.

BACKGROUND: Microbiome therapies (probiotic, prebiotic, and synbiotics) have been proposed as adjuvants in the control of central obesity; however, their results for patients with type 2 diabetes (T2D) remain inconclusive. OBJECTIVE: The aim of this systematic review and meta-analysis was to evaluate the effect of microbiome therapies on central obesity as measured by waist circumference (WC), and to evaluate the effect of microbiome therapies for glycemic parameters (fasting glucose [FPG], fasting insulin [FPI], hemoglobin A1c [HbA1c], and insulin resistance [HOMA1-IR]) in patients with T2D. METHODS: SCOPUS, Pubmed, EBSCO, and LILACS databases were searched for studies that investigated the effect of microbiome therapies on WC up to June 1, 2022. Heterogeneity was determined using Cochran's Q test and quantified using the inconsistency index. The random effects model was used to calculate the pooled difference in means (DM) and 95% confidence intervals (95%CI). Egger's test and Beggs-Muzamar's test were used to assess publication bias. RESULTS: Fifteen reports were included (443 treated and 387 controls). Overall, a significant decrease in WC was found (DM = -0.97 cm; 95% confidence interval [95%CI] = -1.74 to -0.20; P = 0.014); however, when stratified by type of microbiome therapy, only probiotics significantly decreased WC (DM = -0.62 cm; 95%CI = -1.00 to -0.24; P = 0.002). No effect was observed for prebiotics and synbiotics. With respect to glycemic parameters, HbA1c, FPG, and HOMA1-IR significantly decrease with microbiome therapies (P ≤ 0.001). When stratified by the type of therapy, for probiotic treatments, HbA1c, FPG, and HOMA1-IR scores decrease (P < 0.001). For prebiotic treatments, HbA1c and FPG (P ≤ 0.001) levels decrease, whereas FPI increased (P = 0.012). Synbiotic treatments were only associated with an increase in FPI (P = 0.031). CONCLUSION: Findings indicate that using probiotics alone improved WC in patients with T2D. Both probiotics and prebiotics decreased HbA1c and FPG; however, prebiotics and synbiotics resulted in an increase in FPI. The formulation of the therapy (single vs multi) had no difference on the effect.

Corrigendum: Maternal and offspring sugar consumption increases perigonadal adipose tissue hypertrophy and negatively affects the testis histological organization in adult rats.

[This corrects the article DOI: 10.3389/fcell.2022.893099.].

Disordered Glucose Levels Are Associated with Xanthine Oxidase Activity in Overweight Type 2 Diabetic Women.

Oxidative stress plays an important role in vascular complications observed in patients with obesity and Type 2 Diabetes (T2D). Xanthine oxidase (XO) breaks down purine nucleotides into uric acid and contributes to the production of reactive oxygen species (ROS). However, the relationship between XO activity and glucose homeostasis in T2D subjects with obesity is unclear. We hypothesized that disordered glucose levels are associated with serum XO activity in overweight women and men with T2D and without hyperuricemia. We studied serum XO activity in women and men with and without T2D. Our results show that serum XO activity was greater in T2D patients with body mass index (BMI) ≥ 25 kg/m2 than in those with BMI < 25 kg/m2 (p < 0.0001). Sex-based comparative analyses of overweight T2D patients showed that serum XO activity correlated with homeostasis model assessment of ß-cell function (HOMA-ß), fasting plasma glucose (FPG), and hemoglobin A1C in overweight T2D women but not in overweight T2D men. In addition, as compared to overweight T2D men, women had higher high-sensitivity C-reactive protein (hs-CRP) levels. However, overweight T2D men had higher XO activity and uric acid levels than women. Our results suggest that XO activity is higher in overweight T2D patients, especially in men, but is more sensitive to disordered glucose levels in overweight women with T2D.

Histomorphological testicular changes and decrease in the sperm count in pubertal rats induced by a high-sugar diet.

BACKGROUND: During childhood and adolescence, excessive food consumption stimulates adipose tissue expansion promoting overweight in humans, and mice. A high-sucrose diet is related to obesity and metabolic syndrome. Infertility is commonly related to these pathologies. We aim to evaluate possible histomorphological testicular changes induced by a high-sucrose diet on sperm count during the post-weaning period. METHODS: Wistar male rats aged 21 days, weaned, were randomly assigned into two groups: control (fed and hydrated normally) and sugar group (fed normally but hydrated with a solution containing 30% of diluted sucrose during 30 days). At the pubertal age of 51 days, animals were killed and blood samples were taken to measure testosterone and leptin. Testicles were collected and gonadal adipose tissue and semen samples from the epididymis were excised. Testicle samples were used for morphological description using H&E staining, as well as to quantify the triacylglycerol content and the lactate dehydrogenase (LDH) expression. Semen samples were used to assess motility, viability, and sperm count. RESULTS: The sugar group presented an increase in the testicular weight, but a reduction in the cross-sectional area of seminiferous tubules. Moreover, disorganization of Sertoli cells and spermatogonia, an increase in the LDH expression within the entire seminiferous tubule, and a reduced sperm count and spermatozoid motility were found. These alterations were accompanied by high serum levels of testosterone and leptin. CONCLUSIONS: Our results indicate strong damage of testis by sugar consumption during early life that may lead to the onset of infertility in adulthood.

High-sucrose diet potentiates hyperaldosteronism and renal injury induced by stress in young adult rats.

Analyze the effect of stress and high-sucrose diet on serum aldosterone levels and the morphometric characteristics of the kidney in young adult rats. Wistar male rats aged 21 days old weaned were randomly assigned into four groups: control (C), stressed (St), high-sucrose diet (S30), and chronic restraint stress plus a 30% sucrose diet (St + S30). Rats were fed with a standard chow and tap water ad libitum (C group) or 30% sucrose diluted in water (S30 group) during eight weeks. The St and St + S30 groups were subject to restraint stress (1-hour daily in a plastic cylinder, 5 days per week), four weeks before euthanasia. At 81 days old, all animals were killed and blood samples and kidneys were collected. Stressed rats had an increase in the serum aldosterone and renal triacylglycerol, a decrease in the area of the renal corpuscle, glomeruli, proximal tubules, and aquaporin 2 expressions with loss of glomeruli. For its part, the high-sucrose diet decreased the area of the renal corpuscle, glomeruli, and aquaporin 2 expressions in the cortex. The combination of stress and high- sucrose diet maintained similar effects on the kidney as the stress alone, although it induced an increase in the creatinine levels and renal glycogen. Our results showed that chronic stress induces hyperaldosteronism and kidney injury. The intake of a high-sucrose diet may potentiate the renal injury promoted by stress.

Hypothyroidism modifies differentially the content of lipids and glycogen, lipid receptors, and intraepithelial lymphocytes among oviductal regions of rabbits.

Hypothyroidism affects the content of triacylglycerol (TAG), total cholesterol (TC), oxidized lipids, glycogen, and infiltration of immune cells into the ovary and uterus. This study aimed to analyze the impact of hypothyroidism on the lipid content of different regions of the oviduct. Control (n = 6) and hypothyroid (n = 6; 10 mg/kg/day of methimazole in the drinking water for 30 days) adult rabbits were used. In the fimbriae/infundibulum (FIM/INF), ampulla, (AMP), isthmus (IST), and utero-tubal junction (UTJ), the TAG and TC concentrations, presence of oxidized lipid, relative expressions of perilipin A (PLIN A), peroxisome proliferator-activated receptor γ (PPARγ), CAAT/enhancer-binding protein α (C/EBPα), and farnesoid X receptor (FXRα) were analyzed. The content of glycogen and glycans, as well as the infiltration of lymphocytes, were also quantified. In the FIM/INF, hypothyroidism reduced the content of TC, expression of C/EBPα, and presence of glycans while increased the number of intraepithelial lymphocytes. In the AMP and IST-UTJ regions, hypothyroidism increased the content of TAG, oxidized lipids, expression of PPARγ, and glycogen content but decreased the expression of PLIN-A. The FXRα expression in secretory cells of IST-UTJ was higher in the hypothyroid rabbits compared to controls. Additionally, hypothyroidism reduced the C/EBPα expression and the number of intraepithelial lymphocytes in the AMP and IST-UTJ regions, respectively. We demonstrated that the effect of hypothyroidism depends on the oviductal region, possibly associated with different physiological functions specific to each region. These alterations may be related to infertility, tubal disturbances, and ectopic pregnancy observed in hypothyroid women.

Inferring lanosterol functions in the female rabbit reproductive tract based on the immunolocalization of lanosterol 14-demethylase and farnesoid beta-receptor.

Female reproductive organs have de novo synthesis of cholesterol. Some sterol molecules, intermediaries in the cholesterol synthesis, have important paracrine/autocrine actions. Lanosterol binds to the farnesoid beta-receptor (FXRß), a molecule widely expressed in the ovaries, suggesting that it may play a role in reproduction. Up to date, we know little about lanosterol functions across female reproductive organs. We described immunolocalized lanosterol 14-demethylase (LDM or CYP51A1), responsible for catalyzing the conversion of lanosterol in cholesterol, and FXRß in the ovary, oviduct, uterus, and vagina of virgin and pregnant rabbits. In virgin rats, we found CYP51A1 and FXRß immunoreactivity was found in all ovarian follicles, epithelial cells, stroma, and Graafian follicles. Also, the epithelium and stroma, as well as the smooth muscle of the oviduct, vagina, and uterus showed CYP51A1 and FXRß immunoreactivity. In pregnant dams, we observed the presence of CYP51A1 and FXRß immunoreactivity in the corpora lutea, giant uterine cells, and trophoblastic cells. The presence of CYP51A1 and FXRß support that lanosterol participates in diverse reproductive processes, including follicular maturation, transport of gametes and zygote, implantation of blastocyst, lubrication, and contraction of the vagina, secretion of female prostate, and control of delivery mediated by pelvic muscles contraction.

Morphometric changes and AQP2 expression in kidneys of young male rats exposed to chronic stress and a high-sucrose diet.

OBJECTIVE: Consumption of a cafeteria-like diet and chronic stress have a negative impact on kidney function and morphology in adult rats. However, the interaction between chronic restraint stress and high-sucrose diet on renal morphology in young rats is unknown. A high-sucrose diet does not modify serum glucose levels but reduces serum corticosterone levels in stressed young rats, in this way it is confusing a possible potentiate or protector effect of this diet on kidney damage induced by stress. METHODS: Wistar male rats at 4 weeks of age were randomly assigned into 4 groups: control (C), stressed (St), high-sucrose diet (S30), and chronic restraint stress plus a 30% sucrose diet (St + S30). Rats were fed with a standard chow and tap water (C group) or 30% sucrose diluted in water (S30 group). Chronic restraint stress consisted of 1-h daily placement into a plastic cylinder, 5 days per week, and for 4 weeks. RESULTS: Stressed rats exhibited a low number of corpuscles, glomeruli, high number of mesangial cells, major deposition of mesangial matrix and aquaporin-2 protein (AQP-2) expression, and low creatinine levels. Meanwhile, high-sucrose diet ameliorated AQP-2 expression and avoided the reduction of creatinine levels induced by chronic stress. The combination of stress and high-sucrose diet maintained similar effects on the kidney as stress alone, although it induced a greater reduction in the area of proximal tubules. CONCLUSIONS: Our results show that both chronic stress and a high-sucrose diet induce histological changes, but chronic stress may generate an accelerated glomerular hypertrophy associated with functional changes before puberty.

Interactive effects of chronic stress and a high-sucrose diet on nonalcoholic fatty liver in young adult male rats.

Glucocorticoids have been implicated in nonalcoholic fatty liver diseases (NAFLD). The influence of a palatable diet on the response to stress is controversial. This study explored whether a high-sucrose diet could protect from hepatic steatosis induced by chronic restraint stress in young adult rats. Male Wistar rats aged 21 days were allocated into four groups (n = 6-8 per group): control, chronic restraint stress, 30% sucrose diet, and 30% sucrose diet plus chronic restraint stress. After being exposed to either tap water or sucrose solution during eight weeks, half of the rats belonging to each group were subject or not to repeated restraint stress (1 h per day, 5 days per week) during four weeks. Triacylglycerol (TAG), oxidative stress, activity of 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD-1), infiltration of immune cells, and glycogen amount in the liver were quantified. Serum concentrations of corticosterone and testosterone were also measured. The stressed group showed normal serum concentrations of corticosterone and did not have hepatic steatosis. However, this group showed increased glycogen, inflammation, mild fibrosis, oxidative stress, and a high activity of 11ß-HSD-1 in the liver. The group exposed to the high-sucrose diet had lower concentrations of corticosterone, hepatic steatosis and moderate fibrosis. The group subject to high-sucrose diet plus chronic restraint stress showed low concentrations of corticosterone, hepatic steatosis, oxidative stress, and high concentrations of testosterone. Thus, restraint stress and a high-sucrose diet each generate different components of nonalcoholic fatty liver in young adult rats. The combination of both the factors could promote a faster development of NAFLD.

Role of Estrogens in the Size of Neuronal Somata of Paravaginal Ganglia in Ovariectomized Rabbits.

We aimed to determine the role of estrogens in modulating the size of neuronal somata of paravaginal ganglia. Rabbits were allocated into control (C), ovariectomized (OVX), and OVX treated with estradiol benzoate (OVX + EB) groups to evaluate the neuronal soma area; total serum estradiol (E2) and testosterone (T) levels; the percentage of immunoreactive (ir) neurons anti-aromatase, anti-estrogen receptor (ERα, ERß) and anti-androgen receptor (AR); the intensity of the immunostaining anti-glial cell line-derived neurotrophic factor (GDNF) and the GDNF family receptor alpha type 1 (GFRα1); and the number of satellite glial cells (SGCs) per neuron. There was a decrease in the neuronal soma size for the OVX group, which was associated with low T, high percentages of aromatase-ir and neuritic AR-ir neurons, and a strong immunostaining anti-GDNF and anti-GFRα1. The decrease in the neuronal soma size was prevented by the EB treatment that increased the E2 without affecting the T levels. Moreover, there was a high percentage of neuritic AR-ir neurons, a strong GDNF immunostaining in the SGC, and an increase in the SGCs per neuron. Present findings show that estrogens modulate the soma size of neurons of the paravaginal ganglia, likely involving the participation of the SGC.

Consumption of sucrose from infancy increases the visceral fat accumulation, concentration of triglycerides, insulin and leptin, and generates abnormalities in the adrenal gland.

Consumption of sugar-sweetened beverages promotes the development of metabolic syndrome (MetS) and type 2 diabetes mellitus in humans. One factor related to the appearance of MetS components is the dysfunction of the adrenal gland. In fact, the experimental generation of hyperglycemia has been associated with morphological and microvascular changes in the adrenal glands of rats. We hypothesized that high sucrose consumption from infancy promotes histological disruption of the adrenal glands associated with the appearance of metabolic syndrome indicators. Male Wistar rats were separated at weaning (21 days old) into two groups: free access to tap water (control group, C) or 30 % sucrose diluted in water (sugar-fed group). After 12 weeks, high sucrose consumption promoted an increase in visceral fat accumulation, adipose cell number, and insulin resistance. Also, a rise in the concentration of triglycerides, very low-density lipoprotein, insulin and leptin was observed. In control rats, a histomorphometric asymmetry between the right and left adrenal glands was found. In the sugar-fed group, sucrose consumption produced a major change in adrenal gland asymmetry. No changes in corticosterone serum level were observed in either group. Our results suggest that a high sucrose liquid-diet from early life alters the morphology of adrenocortical zones, leading to MetS indicators.

High Sucrose Intake Ameliorates the Accumulation of Hepatic Triacylglycerol Promoted by Restraint Stress in Young Rats.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disorder. Stress promotes the onset of the NAFLD with a concomitant increment in the activity of the hepatic 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD-1). However, the interaction between the stress and a carbohydrate-enriched diet for the development of NAFLD in young animals is unknown. In the present study, we evaluated the impact of chronic stress on the hepatic triacylglycerol level of young rats fed or not with a high sucrose-diet. For doing this, 21-day old male Wistar rats were allocated into 4 groups: control (C), chronic restraint stress (St), high-sucrose diet (S30), and chronic restraint stress plus a 30 % sucrose diet (St + S30). Chronic restraint stress consisted of 1-hour daily session, 5 days per week and for 4 weeks. Rats were fed with a standard chow and tap water (C group) or 30 % sucrose diluted in water (S30 group). The St + S30 groups consumed less solid food but had an elevated visceral fat accumulation in comparison with the St group. The St group showed a high level of serum corticosterone and a high activity of the hepatic 11ß-HSD-1 concomitantly to the augmentation of hepatic steatosis signs, a high hepatic triacylglycerol content, and hepatic oxidative stress. Conversely, the high-sucrose intake in stressed rats (St + S30 group) reduced the hepatic 11ß-HSD-1 activity, the level of serum corticosterone, and the hepatic triacylglycerol content. Present findings show that a high-sucrose diet ameliorates the triacylglycerol accumulation in liver promoted by the restraint stress in young male rats.