Ultrastructural study of the ventral lobe of the prostate of mice with streptozotocin induced diabetes (C57BL/6J)

Morphological and functional changes caused by diabetes in the accessory sex organs and especially the prostate have been reported by several investigators. The aim of the present study was to examine the possible deleterious effects of experimentally induced diabetes on the secretory epithelium of the ventral prostate of mice. Sixteen adult male C57BL/6J mice were divided into two groups. The diabetic group received a streptozotocin injection of 75 mg/kg, while the control group received only 0.1 ml citrate buffer, i.p. After 30 days, the diabetic state was ascertained, the animals were sacrificed and the ventral lobe of the prostate was collected for histological and ultrastructural examination. The results showed reduction in glandular epithelium cell height, increased numbers of cytoplasmic vacuoles and thickening of the extracellular matrix. In conclusion, experimental diabetes has harmful effects on the secretory epithelial cells of the ventral lobe of the prostate of mice.     

Experimental alcoholism and pathogenesis of prostatic diseases in UChB rats

Previous studies have shown that long-term alcohol treatment has negative effects on prostatic stromal-epithelial interaction. Thus, the aim of the present study was to analyze the histochemical, immunohistochemical and ultrastructural alterations that occur in the prostatic stroma and epithelium of rats submitted to chronic alcohol ingestion and alcohol abstinence, as well as to establish the relationship between these changes and prostatic diseases. Thirty male rats (10 Wistar and 20 UChB rats) were divided into three experimental groups: the control group received tap water, the alcoholic group received ethanol diluted to 10 degrees G.L. for 150 days, and the abstinent group received the same liquid diet as the alcoholic group up to 120 days of treatment and only tap water for 30 days thereafter. At the end of treatment, all animals were sacrificed and the ventral lobe of the prostate was removed and processed for histochemical, immunohistochemical and ultrastructural analyses. In addition, plasma testosterone levels were measured. The results showed prostatic intraepithelial neoplasia, infolding of the epithelium towards the stroma, stromal hypertrophy and the presence of inflammatory cells in alcoholic animals. In the abstinent group, alterations were noted mainly in the stromal area. In conclusion, ethanol triggers alterations in prostatic epithelial and stromal compartments, affecting the stromal microenvironment and predisposing the organ to pathological processes.     

Influence of hormonal replacement on the ventral lobe of the prostate of rats (Rattus norvegicus albinus) submitted to chronic ethanol treatment

The harmful influence of the chronic alcohol ingestion on the male reproductive system leads to important alterations including hypogonadism and feminization, besides the morphological and functional disorganization of the different sexual glands. So, the aim of this study was to analyse the structural changes on the ventral lobe of the prostate of rats with hormonal replacement associated to chronic alcohol ingestion. A total of 30 rats (Rattus norvegicus albinus) was divided into three groups: control-received water; alcoholic-received ethanol diluted to 20% and hormone-treated alcoholic-received ethanol diluted to 20% associated with the administering of testosterone (5mg/kg of weight) during the last 30 days of treatment. After 150 days of treatment, the animals were sacrificed, the prostate removed and submitted to transmission and scanning electron microscopies, histochemical analysis for acid phosphatase, testosterone level and stereologic analysis. In the alcoholic group the results demonstrated reduction of the total cellular volume and disorganization of the organelles involved in the secretory process. It was characterized a partial recovery of the cellular volume after treatment with testosterone. It was concluded that the ethanol impaired the cellular morphology and the hormonal replacement by itself did not bring about efficient remodeling of the organelles responsible for the secretory process.     

IGFR-I expression and structural analysis of the hard palatine mucosa in an ethanol-drinking rat strain (UChA and UChB)

The study analyzed the effects of chronic alcohol ingestion on the ultrastructure of the lining epithelium of the hard palatine mucosa of rats UChA and UChB (lines with voluntary alcohol consumption) in order to contribute to the understanding of the consequences of alcohol abuse for the morphology of the digestive system. Thirty female adult animals aged 120 days were divided into three experimental groups. (1) Ten UChA rats (genetically low ethanol consumer) with voluntary intake of 10% v/v (5.45 g/kg/day) ethanol solution and water. (2) Ten UChB (genetically high ethanol consumer) rats with voluntary intake of 10% v/v (7.16 g/kg/day) ethanol solution and water. (3) Ten Wistar rats with voluntary ad libitum water intake (control group). Both groups received Nuvital pellets ad libitum. The IGFR-I expression was intense in both experimental groups. The epithelial cells of the alcoholic rats UChA and UChB showed many alterations such as the presence of lipid droplets, altered nuclei, nuclei in corneum layer and disrupted mitochondria. It was concluded that ethanol intake induces ultrastructural lesions in the hard palatine mucosa.     

Stereology and ultrastructure of the seminal vesicle of C57/BL/6J mice following chronic alcohol ingestion

Excessive alcohol consumption causes metabolic changes and pathologic alterations in testes and accessory sex organ in different animal species. The aim of the present study was to evaluate the macroscopic, histologic and ultrastructural alterations provoked by chronic ingestion of different ethanol concentrations over increasing periods of time on the secretory epithelium of the seminal vesicle of C57/BL/6J mice in using stereological methods. Sixty male adult mice were divided into three experimental groups: Control, Alcoholic 25% and Alcoholic 35%, respectively, receiving tap water and tap water containing ethanol diluted to 25 and 35 degrees Gay Lussac. All mice were fed with the same solid diet. After 150 and 250 days of treatment the animals were sacrificed and the seminal vesicles were collected and processed for light and transmission electron microscopy. The cellular, cytoplasmic and nuclear volumes and the area density of autophagic and secretory vacuoles were measured. The histologic alterations observed in the alcoholic mice consisted of a reduction in epithelial size and cell volume, with maintenance of the same nuclear and cytoplasmic ratio as verified in the control groups. The ultrastructural alterations were: increased density of dense body area, decreased density of secretory granule area, and dilated rough endoplasmic reticulum and Golgi cisternae. We conclude that chronic ethanol ingestion causes depleting morphologic alterations in the epithelial cells of the seminal vesicle and negatively affects the secretory process of this gland.     

Morphometric and morphological features of the ventral prostate in rats submitted to chronic nicotine and alcohol treatment

Clinical studies analyzing simultaneous nicotine-alcohol use by patients showed important alterations in various organic systems such as: respiratory, digestory, and genital. Also, the prostatic morphology and physiology have been analyzed, specially due to large occurrence of prostatic diseases. Then, this work aimed at determining the structure and ultrastructure of the prostatic stroma and epithelium, as well as the stroma epithelium interactions from rats submitted to simultaneous long-term alcohol-nicotine treatment. A total of 40 male rats were divided into four groups: control group (10 animals) received tap water; alcoholic group (10 animals) received diluted 10% Gay Lussac ethanol; nicotine group (10 animals) received a 0.125mg/100g of body weight dose of nicotine injected subcutaneosly on a daily basis; nicotine-alcohol group (10 animals) received simultaneous alcohol and nicotine treatment. After 90 days of treatment, the animals were sacrificed and samples from the ventral lobe of the prostate were collected and processed for transmission electron and light microscopies. The results showed atrophied epithelium; prostatic intra-epithelial neoplasia; dilated cisterns of the granular endoplasmic reticulum, large amounts of collagen fibers besides inflammatory cells, specially in the alcoholic and nicotine-alcohol groups. Therefore, it could be concluded that the association between alcohol and nicotine caused the impairment of the prostatic secretory process. Moreover, this association is related to prostatic pathogenesis, which could lead to late glandular malignancy.     

Acute and chronic dose of alcohol affect the load carrying capacity of long bone in rats

In the present investigation, the effects of acute and chronic dose of alcohol were evaluated on mechanical properties of long bones of Sprague Dawley rats. In "acute study", 18 animals were divided into three groups containing six animals each, i.e. Group A: control animals, normal saline was given to them intraperitoneally for the period of 5 days; Group B: treated animals, given 20% (v/v) absolute alcohol and Group C: treated animals, given 30% (v/v) absolute alcohol, by same route and time duration. In "chronic study", also, 18 animals were divided into three groups containing six animals each, i.e. Group A: control animals, normal saline was given to them intraperitoneally for the period of 6 weeks; Group B: treated animals, given 20% (v/v) absolute alcohol and Group C: treated animals, given 30% (v/v) absolute alcohol by same route and time duration. A significant increase was observed in bone weight of animals taking 20% alcohol but there was decrease in the same for 30% alcohol in case of acute study. For chronic study, there was a decrease in bone weight for both treated groups. During acute study, breaking strength of bone was increased in case of 20% alcohol administration but a slight decrease was shown in the same for 30% alcohol group as compared to control animals. Breaking strength of long bone in the case of chronic study was decreased in case of both groups taking alcohol, i.e. 20% and 30%. The present document is useful in understanding the functional load carrying capacity of bone during alcoholism.     

Alcoholism and coagulating gland: Androgen and insulin like growth factor-1 receptor features

The aim of this work was to characterize the structural and molecular changes in the coagulating gland from rats submitted to long-term alcohol treatment, as well as the possibility of recovery of these parameters after interrupting the alcohol administration. Ten Wistar and twenty UChB rats were divided into: Control group received tap water; Alcoholic group received 10% (v/v) ethanol daily for 150 days; and Abstinent group, received 10% (v/v) ethanol daily for 120 days and then tap water like the control for another 30 days. After 150 days, samples from the coagulating glands were processed for morphological and immunohistochemical analyses. The results showed atrophied epithelium and hypertrophied stroma, especially in the alcoholic group. Intensed androgen receptor (AR) immunolocalization was verified in the epithelium and weak in the stroma of the control group in relation to the other groups. Intensed insulin-like growth factor receptor-1 (IGFR-1) immunolocalization was verified in the stroma of the alcoholic and abstinent groups. Thus, it could be concluded that the excessive alcohol consumption caused morphological and molecular changes in the coagulating gland, characterizing the inverse relation of AR and IGFR-1 localization. The alcohol was an important factor in cellular mitosis occurrence, which could be fundamental element involved in glandular lesions.     

Structural evaluation of the effects of chronic ethanol ingestion on the testis of Calomys callosus

Chronic exposure to ethanol may results in pathophysiologic changes in cellular function. The present work was designed to investigate the morphology of testis submitted to experimental ethanol ingestion. Experimental animals were divided into two groups. The control group (n = 23) received a solid diet and tap water and the alcoholic group (n = 23) received the same solid diet and ethanol P.A. diluted 20% in water (v/v). After 120 days of treatment, all animals were anesthetized, weighed and sacrificed. Testosterone and luteinizing hormone levels in serum were lower in the alcoholic group than in the control group. Histological and ultrastructural alterations were observed in the testicular alcoholic germinative cells like enormous spaces, lipid droplets accumulation, digestive vacuoles, irregular diameter of the seminiferous tubules and interstitial dilated blood vessels. It was concluded that 20% ethanol provokes lesions on the testis germinative epithelium probably inducing gonadal dysfunction.     

Chronic effects of alcohol on the epithelium of the small intestine using two experimental models

The effects of chronic alcohol ingestion on the rat small intestinal epithelium have been measured using two different experimental models, viz. either a solid diet plus alcohol or a liquid diet plus alcohol. All experimental animals consumed adequate quantities of food and exhibited a normal growth rate; thus the recorded effects were unlikely to be associated with malnutrition. After a period of 24 days, there were significant decreases in the jejunal villus cell population in both groups of experimental animals and corresponding increases in the crypt cell populations; no such changes were found in the ileum. Alcohol had no significant effect on the crypt cell production rate and the growth fraction at either of these two sites in the intestine, and thus the major effect of alcohol was to cause a lengthening of the cell cycle time in the jejunal crypts.     

酒精对丙酸睾酮诱导的小鼠前列腺增生的作用

目的：探讨酒精对丙酸睾酮引起的小鼠前列腺增生（BPH）的作用及其生殖毒性。方法：成年雄性昆明系小鼠70只随机分为空白对照组（Control）、阴性对照组（Negative control,sc大豆油25 mg/（kg·d）,ig蒸馏水7.5 ml/（kg·d）,连续处理7 d、21 d）、酒精7 d和21 d组（AL7和AL21,ig 50°白酒7.5 ml/（kg·d）,连续处理7 d、21 d）,丙酸睾酮7 d和21 d组（TP7和TP21,sc丙酸睾酮注射液25 mg/（kg·d）,连续处理7 d、21 d）,丙酸睾酮+酒精7 d组（TP+AL7,sc丙酸睾酮注射液25 mg/（kg·d）,ig50°白酒7.5 ml/（kg·d）,连续处理7 d）,每组10只。末次处理24 h后处死小鼠,计算小鼠前列腺和睾丸系数,检测精子参数,测定睾丸和前列腺组织中自由基水平、抗氧化能力,观察前列腺组织病理学变化。结果：与对照组、TP7 d组、AL7和AL21 d组相比,TP+AL7 d组的前列腺系数显著提高、精子数量和质量显著降低、前列腺和睾丸MDA含量显著升高、SOD和GPx酶活力显著下降（P均< 0.05）;与TP21 d组相比,TP+AL7 d组的前列腺系数无显著差别（P>0.05））。结论：丙酸睾酮和酒精共同处理7 d就可以达到典型的前列腺增生（BPH）状态,并引起睾丸及精子的损伤,导致生殖系统的氧化应激反应增强,说明酒精对丙酸睾酮引起的小鼠前列腺增生有明显的促进作用。     

Morphometric analysis of the rat ventral prostate and seminal vesicles during prepubertal development: effects of neonatal treatment with estrogen

In a morphometric study on the ventral prostate and seminal vesicles in the rat, we investigated the changes in fibromuscular stroma, glandular epithelium, and glandular lumen. Animals were studied at 15, 30 and 45 days of age. The rapid prepubertal growth started earlier in the ventral prostate than in seminal vesicles. In addition, the effects of neonatal administration of estrogens on the different tissue compartments were studied, comparing rats that had been castrated and/or treated with estrogen at birth to intact animals at 15 days of age. Estrogens caused a decrease in the volume of the glandular epithelium and increased the volume of the fibromuscular stroma in both ventral prostate and seminal vesicles. Castration partially abolished the estrogen-induced growth of the stroma, which suggests that the growth is dependent on testicular factors. The difference in proportion of the fibromuscular stroma between the two glands is evidence that the size of the whole seminal vesicles has increased whereas the size of the ventral prostate has decreased.     

Ultrastructural and proliferative features of the ventral lobe of the prostate in non-obese diabetic mice (NOD) following androgen and estrogen replacement associated to insulin therapy

Diabetes causes harmful effects on prostatic function. Thus, the aims of this study were to characterize morphological and proliferative features of the prostate of diabetic mice after long-term glycemic control and testosterone and estrogen replacement. A total of 48 mice (Nod and BALBc) were used. After 20 days in a diabetic state, the mice were divided into six groups: the control group received a 5 mL/kg dose of peanut oil; the diabetic group received the same treatment as the control group; the diabetic-insulin group received 4 IU doses of insulin; the diabetic-testosterone group received a 5 mg/kg dose of testosterone cypionate; the diabetic-estrogen group received a 25 μg/kg dose of 17β-estradiol; the diabetic-insulin-testosterone-estrogen group received insulin, testosterone and estrogen at the same concentration as the other groups. After 20 days, the ventral lobe was processed for morphological and immunological analyses. The results showed structural disorganization, which was more intense in the diabetic group than in the other groups. The diabetic state showed a proliferation and apoptosis rate that was two times higher than that found in the control group. To conclude, diabetes disturbed the prostatic secretory activity and the association of insulin, testosterone and estrogen was crucial for glandular structural restoration, characterizing the complex activity of the prostate. The imbalance verified between the proliferation process and apoptosis in diabetic mice showed diabetes to be a triggering factor for prostatic pathogenesis.     

Effect of ethyl alcohol on plasma testosterone level in mice

The effect of ethanol ingestion on testicular steroidogenesis in mice was evaluated by measuring plasma testosterone level. Four groups of CBA/J male mice were treated with one of the following doses of ethanol: 1.240, 0.620, 0.310 or 0.155 g ethanol/Kg body weight. A control group was given water. The data showed no effect of the treatment on testicular weight. The concentration of testosterone in the plasma was significantly reduced in animals treated with alcohol. There was also a significant relationship between the dose of alcohol and the plasma testosterone level, with the decrease in testosterone being from 2 to 18 fold in various groups.     

The EZC-prostate model: noninvasive prostate imaging in living mice

Recently, progress in the development of prostate-specific promoters and high resolution imaging techniques has made real-time monitoring of transgenic expression possible, opening a vista of potentially important in vivo models of prostate disease. Herein, we describe a novel prostate reporter model, called the EZC-prostate model that permits both ex vivo and in vivo imaging of the prostate using a sensitive charge-coupled device. Firefly luciferase and enhanced green fluorescent protein were targeted to the prostate epithelium using the composite human kallikrein 2 (hK2)-based promoter, hK2-E3/P. In EZC-prostate mice, the ventral and dorsal/lateral prostate lobes were brilliant green under fluorescence microscopy, with expression localized to the secretory epithelium. In contrast, enhanced green fluorescent protein was undetectable in the anterior lobes of prostate, seminal vesicles, testes, liver, lung, and brain. The kinetics of luciferase activity in intact and castrated living mice monitored with the IVIS charge-coupled device-based imaging system confirmed that firefly luciferase expression was largely prostate restricted, increased with age up to 24 wk, and was androgen dependent. Decreases in reporter expression after 24 wk may reflect well known, age-related decreases in androgen signaling with age in humans. Ex vivo imaging of microdissected animals further confirmed that the luminescence detected in living mice emanated predominately from the prostate, with minor signals originating from the testes and cecum. These data demonstrate that the hK2-E3/P promoter directs strong prostate-specific expression in a transgenic mouse model. Multigenic models, generated by crosses with various hyperplastic and neoplastic prostate disease models, could potentially provide powerful new tools in longitudinal monitoring of changes in prostate size, androgen signaling, metastases, or response to novel therapies without sacrificing large cohorts of animals.     

Intracellular localization of prostatic binding protein (PBP) in rat prostate by light and electron microscopic immunocytochemistry

Summary Extra- and intracellular distribution of Prostatic Binding Protein (PBP) was studied in the different genital organs of the male rat by immunocytochemistry at the light and electron microscopic levels. PBP was extracted from cytosols of rat ventral prostate and used for immunization of rabbits. The specificity of the antiserum raised was tested by western blotting and immunoelectrophoresis. From the different fixatives tested for optimal structural and antigenic preservation of the ventral prostate a mixture containing 2.5% paraformaldehyde, 0.5% glutaraldehyde and 0.5% CaCl₂ in cacodylate buffer, 0.05 M, pH 7.3 was selected. Using the immunofluorescence technique and the unlabeled antibody enzyme method PBP-immunoreactivity was detected at the light microscopic level in the luminal secretions of the ventral prostate. No reaction was observed with the seminal vesicle, the coagulating gland, the dorsal and lateral prostates, the epididymis and the testis. Intracellular secretory granules reacting with PBP antiserum were exclusively found in the secretory cells of the ventral prostate. Insufficiently fixed cells showed a diffuse generalized reaction of the cytoplasm indicating a leakage of the antigen from the secretory granules. Such artifacts were common in tissue sections processed with the preembedding-staining procedure. At the ultrastructural level therefore mostly the postembedding staining method was performed using both the unlabeled antibody enzyme method and the ferritin-labeled immunoglobulin technique in osmicated, Epon-embedded tissue. Labeling with either method was intense in the secretory granules and the condensing vacuoles, while the labeling density of the rough endoplasmic reticulum and the Golgi cisternae was in the background range. Castration experiments showed that secretory material displaying PBP immunoreactivity was retained within the acinar lumen of the gland for several days after castration, but was absent from most secretory cells already by four days after castration. Immunocytochemistry of PBP therefore is a very sensitive method for analysing the secretory activity and its androgen dependence of the prostate of the rat.Supported by a grant from the Deutsche Forschungsgemeinschaft (Au 48/7-6) and a grant from the Cystic Fibrosis Foundation (G 261 A)     

Radioprotection by pretreatment with deuterated water: cytokinetic changes in the small intestine of the mouse

All mice partially deuterated by ingestion of 29% heavy water for 12 days survived whole body gamma irradiation (8.5 Gy) from a 60Co source, whereas 42% of nondeuterated control animals died from bone marrow failure. The incorporation of 3HTdR into enterocytic DNA, as measured by autoradiography and liquid scintillation spectrometry, was used to assess the proliferative activity of small intestinal epithelium. The sequence and the magnitude of changes in tritium activity were in good agreement. Deuteration alone resulted in a reduced proliferative activity of small intestinal crypt epithelium, particularly in the basal cell positions and the first positions of the proliferation compartment. The number of positions occupied by the proliferative compartment and the crypt length were, however, barely affected by deuteration. The radiation-induced depression of DNA synthesis in the proliferative compartment was of similar magnitude in both groups. Crypt epithelium in deuterated mice, however, displayed signs of an accelerated and/or enhanced regeneration. The cytokinetic changes in deuterated animals are consistent with a protective effect for clonogenic intestinal epithelium at the time of irradiation.     

Electroacupuncture inhibits CB1 upregulation induced by ethanol withdrawal in mice

CB1R play a role in alcohol withdrawal and in some effects of acupuncture. Interestingly, acupuncture has been used to alleviate alcohol withdrawal. Here, we investigated electroacupuncture (EA) effects during ethanol withdrawal on CB1R immunoreactivity. Male Swiss mice were daily injected with ethanol (2g/kg, i.p) (EtOH group), for 21 days. EA was performed daily during 4 days of ethanol withdrawal. The stimuli of 2 or 100 Hz were provided in two acupoints combination: Ea1 [(ST-36/Zusanli) and (PC-6/Neiguan)] or Ea2 [(DU-14/Dazhui) and (DU-20/Baihui)]. The specificity of the acupoints were assessed by the inclusion of three additional groups, Ea3 [(ST 25/Tianshu - acupoints used to other non-related disorders)], Sham1 and Sham2 (transdermic stimulation nearly to the respective acupoints). EtOH group were only handled during withdrawal and Saline group was chronically treated with Saline and handled similarly to EtOH group. One day after withdrawal the animals were perfused and their brains processed for immunohistochemistry. There was an increase of CB1R in the prefrontal cortex, striatum, hippocampus, amygdala and ventral tegmental area. The procedures used in the 2HzEa1 and 100HzEa2 groups were the most effective and specific to inhibit this CB1R upregulation. Therefore, EA inhibits CB1R upregulation seen in ethanol withdrawn mice. The specificity of acupoints stimulation depends of the encephalic nuclei, acupoints association and frequency of stimulation.     

Prostatic stromal microenvironment and experimental diabetes

The diabetes causes alterations in various organ systems, including the male accessory sex glands. The prostate is very important in the reproductive process and it is a frequent target of malignant changes. The aim of this work was to demonstrate the histochemical and ultrastructural alterations in the prostate of diabetic animals. Two groups of animals were utilized: control and non-obese diabetic mice (NOD). Twelve days after the characterization of diabetic status the ventral prostate was collected, fixed in Karnovsky and paraformaldehyde, processed for histochemistry and TEM associated to stereology. The results showed reduction of the epithelial area and increasing of the stromal area with muscular and collagen hypertrophy in the prostatic gland. It was characterized the development of prostatic intraepithelial neoplasia, inflammatory processes and dilation of the organelles involved in the secretory process. It was concluded that diabetes besides damaging the reproductive process, affects the glandular homeostasis favoring the development of prostatic pathologies.