Jorge Luiz Alves-Pereira

Molecular and morphometric analysis of the rat ventral prostate injected with leptin

The aim of this study was to evaluate the effect of leptin administration on the ventral prostate lobe of adult rat. Twenty adult male rats were divided into 2 groups: L-animals were daily injected with 50 μL of leptin (8 μg/100 g BW, subcutaneous) for four days and C-animals received the same volume of saline solution. Lipid profile and testosterone serum levels were evaluated. The prostate ventral lobe was processed for histomorphometric analysis. Gene expression of aromatase, androgen, leptin and estrogen receptors isoforms was evaluated by real-time PCR. Cell proliferation was evaluated by PCNA immunohistochemistry. Data were expressed as mean±standard error and analyzed by students t-test. Serum levels of cholesterol (C=39.7±4.2;L=55.2±4.2, mg/dL; P<0.02) increased and testosterone (C=1.6±0.43;L=0.6±0.15, ng/dL; P<0.03) decreased in L group. The histomorphometric analysis showed a reduction in cell density (C=8868±242; L=8211±210, mm(2); P<0.04), in total (C=0.24±0.026; L=0.10±0.009, mm(2); P<0.001) and in the internal acini areas (C=0.16±0.009; L=0.08±0.006, mm(2); P<0.0002). On the other hand, there was an increase in the epithelial height (C=17.3±0.3; L=22.8±0.2, μm; P<0.0001) and in the number of acini (C=7.0±0.2; L=8.7±0.1, mm(2); P<0.0002). The histomorphometric analyses together with PCNA immunohistochemistry results suggest that leptin increases cell proliferation. In relation to the gene expression, leptin treatment increased the expression of all genes, but ER-α, in more than 200 times compared to the expression in C group. In conclusion, in this paper we showed that leptin has a direct effect on the prostate gland of adult rats leading to an increase in proliferation and in the gene expression of aromatase, androgen, leptin and estrogen receptors isoforms that are important for the physiology of the prostate gland.

Collagen I and III and metalloproteinase gene and protein expression in prostate cancer in relation to Gleason score

PURPOSE To evaluate if the expression of metalloproteinase, collagen I and III are related to Gleason score, preoperative PSA and pathological stage in prostate cancer. MATERIALS AND METHODS Our study group included radical prostatectomy specimens of 33 patients with prostatic adenocarcinoma who underwent surgery from 2001 to 2009. Patients were divided into 3 groups: Gleason score=6 (13 patients), Gleason score=7 (10 patients), Gleason score ≥ 8 (10 patients). The control group included prostates of patients submitted to cystoprostatectomy and benign prostatic tissues adjacent to the cancer area. Specific areas of tissues were selected under microscope and further processed for collagen I and III analysis by real time PCR. In addition, 10 deparaffined sections of each group were used to evaluate collagen I, III and metalloproteinase immune expression. The results were correlated with Gleason score, preoperative PSA and pathological stage. RESULTS We found significant difference in both collagen I and III gene expression between benign and tumoral areas in the prostate samples from Gleason score=6 (collagen I=0.4 ± 0.2 vs 5 ± 2.4, p < 0.05; collagen III=0.2 ± 0.06 vs 0.7 ± 0.1, p < 0.05) and Gleason score ≥ 8 (collagen I=8 ± 3.4 vs 1.4 ± 0.8, p < 0.07; collagen III=1.8 ± 0.5 vs 0.6 ± 0.1, p < 0.05). There was no correlation of collagen expression with Gleason score, preoperative PSA or pathological stage. There was a positive correlation between metalloproteinase expression and Gleason score (r(2)=0.47). CONCLUSIONS The positive correlation between metalloproteinase expression and Gleason score suggests that metalloproteinase could be a promising factor to improve Gleason score evaluation. Its expression and regulation do not seem to be related with collagen degradation.

Leptin Regulates Proliferation and Apoptosis in Human Prostate

This paper aimed to evaluate the leptin role on the cellular proliferation and the expression of fibroblast growth factor 2, aromatase enzyme, and apoptotic genes in the human prostate tissue. Methods. Fifteen samples of hyperplasic prostate tissue were divided in four symmetric parts maintained in RPMI medium supplemented with 10% fetal bovine serum, 1 ng/mL of gentamicin, and added with 50 ng/mL leptin (L) or not (C). After 3 hours of incubation, gene expression was evaluated by real time RT-PCR. Cellular proliferation was evaluated by immunohistochemistry for PCNA. Results. The leptin treatment led to an increase cellular proliferation (C = 21.8 ± 0.5; L = 64.8 ± 0.9; P < 0.0001) and in the expression of Bax (C = 0.4 ± 0.1; L = 0.9 ± 0.2; P < 0.05) while Bcl-2 (C = 19.9 ± 5.6; L = 5.6 ± 1.8; P < 0.05), Bcl-x (C = 0.2 ± 0.06; L = 0.07 ± 0.02; P < 0.05), and aromatase expressions (C = 1.9 ± 0.6; L = 0.4 ± 0.1; P < 0.04) were significantly reduced. Conclusion. Leptin has an important role in maintaining the physiological growth of the prostate since it stimulates both cellular proliferation and apoptosis, with the decrement in the aromatase gene expression.

Origin and Antimeric Distribution of the Femoral Nerves in New Zealand Rabbits

Se estudio el origen y distribucion del nervio femoral de ambos antimeros en 30 conejos neozelandeses, 15 machos y 15 hembras. Los animales fueron recolectados despues de su muerte natural y se fijaron en formaldehido al 10%. En los machos, el nervio femoral se origino a partir de los ramos ventrales del cuarto, sexto y septimo nervios espinales lumbares en siete casos (46,67%); en tres casos (20%) desde los ramos ventrales del quinto, sexto y septimo nervios espinales lumbares; en dos casos (13,33%) desde los ramos ventrales del quinto y sexto nervios espinales lumbares, mientras que en tres animales (n=1 respectivamente), desde los ramos ventrales del quinto nervio espinal lumbar (6,67%), los ramos ventrales del cuarto y quinto nervios lumbares espinales (6,67%) y desde los ramos ventrales del quinto y septimo nervios espinales lumbares. En las hembras, el nervio femoral se origino a partir de los ramos ventrales del cuarto, sexto y septimo nervios espinales en cuatro casos (26,67%); en otros cuatro casos (26,67%) desde los ramos ventrales del quinto, sexto y septimo nervios espinales lumbar, en tres casos (20%) desde los ramos ventrales del sexto y septimo nervios espinales lumbares, en dos casos (13,33%) desde los ramos ventrales del quinto y sexto nervios espinales, y en dos animales (n=1, respectivamente) procedian desde los ramos ventrales del quinto nervio espinal lumbar (6,67%) y de los ramos ventrales del cuarto y septimo nervios espinales lumbares (6,67%). Los nervios femorales en todos los animales estaban distribuidos en diversos ramos de los musculos psoas mayor y menor, cuadriceps femoral sartorios y pectinatos.

Effects of a high energy density diet in the “corpus cavernosum ” of mice

Erectile dysfunction is a common condition that affects men over age 40. It is highly related to obesity. The corpus cavernosum is the most important structure involved in erection. The aim of this study was to evaluate the structure of the corpus cavernosum of mice fed with a high energy density diet (HED). At 3 months of age, male C57BL/6 mice were fed with a HED diet (50% lipids) or standard chow (SC) diet (10% lipids) for 14 weeks. Afterwards, the animals were euthanized and the corpus cavernosum was analyzed through stereology. Statistical significance was calculated by the student’s t-test (p < 0.05). The group fed with HED diet showed higher values of body weight, blood pressure and higher rates of cholesterol, triglycerides, and glucose from the second week to the end of the experiment. The HED group showed a significant increase in the connective tissue (15%) and a decrease in smooth muscle fibers (41%). The testosterone concentration in the HED group was 63% lower than in SC animals. Animals fed with a HED presented reduced testosterone serum levels and morphological changes on the corpus cavernosum, which may be related to erectile dysfunction.

The testis of the mice C57/BL6 offspring in adulthood have alterations due to maternal caffeine consumption

PURPOSE To investigate the effects of the maternal caffeine consumption during pregnancy to adult male testis mice offspring. METHODS Twenty pregnant mice were divided into control group (c) and caffeine group (cf). dams received daily saline or 20 mg/kg of caffeine subcutaneously. Male offspring were monitored daily until 13th week. The testis were used to evaluate both the proliferation (pcna) and apoptosis (bax); leptin receptor (ob-r); aromatase; follicle stimulating hormone (fshr), luteinizing hormone (lhr) and androgen receptors (ar); steroidogenic acute regulatory protein (star); vascular endothelial growth factor (vegf) and estrogen receptors (erα and erβ) by western blotting. Serum concentrations of testosterone, estradiol and leptin were measured. RESULTS There was a significant reduction in food intake and the body mass gain (p<0.05) in the cf ; pcna (p=0.01), fshr (p=0.02), star (p=0.0007), vegf (p=0.009), ar (p=0.03) in the cf. while an increase were note in bax (p=0.01), ob-r (p=0.02), lhr (p=0.04) and in the aromatase (p=0.03) in the cf. only erα and erβ were not changed by maternal caffeine. The serum testosterone levels in the cf offspring were 90% lower than in the c offspring (p=0.04). CONCLUSION Maternal caffeine consumption has a role and alters the testis of the offspring in adulthood.