[Rauwolfia extract inhibits the proliferation of prostate cells in rats with benign prostatic hyperplasia].

OBJECTIVE: To investigate the effects of different concentrations of Rauwolfia extract (RE) on the proliferation of prostate cells in the rat model of benign prostatic hyperplasia (BPH). METHODS: We randomly divided 48 male SD rats into six groups of an equal number, BPH model control, finasteride, low-concentration RE, medium-concentration RE, high-concentration RE and normal control, and established a BPH model in the former five groups by subcutaneous injection of testosterone propionate following castration. We treated the rats of the finasteride and RE groups intragastrically with finasteride solution at 5 mg/kg and RE at 5, 10 and 20 mg/kg respectively, and those of the model control and normal control groups with an equal dose of normal saline, all once a day for 28 consecutive days. Then, we killed all the animals, collected their prostate tissue, obtained the wet weight and volume of the prostate, the prostate index and the contents of serum T and dihydrotestosterone (DHT), observed the morphological changes of the prostate tissue by HE staining, counted the glands in the prostate tissue, measured the intraglandular area, and determined the expressions of PCNA and α-SMA by immunohistochemistry. RESULTS: Compared with the rats of the normal control group, the BPH model controls showed significantly increased wet weight (ï¼»0.923 ± 0.15ï¼½ vs ï¼»1.455 ± 0.52ï¼½ g, P < 0.05), volume (ï¼»1.035 ± 0.29ï¼½ vs ï¼»1.687 ± 0.31ï¼½ ml, P < 0.05) and index of the prostate (ï¼»0.23 ± 0.04ï¼½% vs ï¼»0.37 ± 0.15ï¼½%, P < 0.05), dilation, hyperemia and edema of the prostatic stroma and vessels, and proliferation rate of the prostatic cells, but remarkably decreased number of glands (ï¼»20.35 ± 3.83ï¼½ vs ï¼»12.56 ± 2.58ï¼½, P < 0.05), epithelial thickness (ï¼»39.76 ± 5.20ï¼½ vs ï¼»19.52 ± 1.52ï¼½ µm, P < 0.05) and intraglandular area (ï¼»12.3 ± 1.21ï¼½ vs ï¼»5.96 ± 0.34ï¼½ ×103µm2, P < 0.05). In comparison with the BPH model controls, the animals treated with RE, especially in the high-concentration RE group, exhibited marked decreases in the weight (ï¼»1.455 ± 0.52ï¼½ vs ï¼»0.862 ± 0.31ï¼½ g, P < 0.05), volume ( ï¼»1.687 ± 0.31ï¼½ vs ï¼»0.952 ± 0.28ï¼½ ml, P < 0.05) and index of the prostate (ï¼»0.37 ± 0.15ï¼½% vs ï¼»0.22 ± 0.07ï¼½%, P < 0.05), dramatic improvement in the number of glands (ï¼»12.56 ± 2.58ï¼½ vs ï¼»18.36 ± 1.25ï¼½, P < 0.05), epithelial thickness (ï¼»39.76 ± 5.20ï¼½ vs ï¼»19.04 ± 3.89ï¼½ µm, P < 0.05) and intraglandular area (ï¼»5.96 ± 0.34ï¼½ vs ï¼»10.25 ± 0.98ï¼½ ×103µm2, P<0.05ï¼½, P < 0.05), remarkable down-regulation of the expressions of PCNA and α-SMA, and significant reduction of the contents of serum T (ï¼»19.147 ± 3.214ï¼½ vs ï¼»6.016 ± 1.978ï¼½ ng/ml, P < 0.05) and DHT (ï¼»9.052 ± 0.633ï¼½ vs ï¼»2.532 ± 0.386ï¼½ ng/ml, P < 0.05). CONCLUSION: Rauwolfia extract can inhibit the proliferation of prostate cells and relieve BPH symptoms in a concentration-dependent manner in rats with BPH.

Rauwolfia vomitoria extract suppresses benign prostatic hyperplasia by reducing expression of androgen receptor and 5α-reductase in a rat model.

OBJECTIVE: Herbal medicine is an important therapeutic option for benign prostatic hyperplasia (BPH), a common disease in older men that can seriously affect their quality of life. Currently, it is crucial to develop agents with strong efficacy and few side effects. Herein we investigated the effects of the extract of Rauwolfia vomitoria, a shrub grown in West Africa, on BPH. METHODS: Rats with testosterone-induced BPH were treated with R. vomitoria. Prostates were histologically analyzed by Hematoxylin and eosin staining. Proliferation index and the expression levels of androgen receptor and its associated proteins were quantified through immunohistochemistry and immunoblotting. Androgen receptor target genes were examined by quantitative real-time polymerase chain reaction. The sperm count and body weight of rats were also measured. RESULTS: The oral administration of R. vomitoria extract significantly reduced the prostate weight and prostate weight index in BPH rats, supported by the decreased thickness of the prostate epithelial layer and increased lumen size. Similar effects were observed in the BPH rats treated with the reference drug, finasteride. R. vomitoria extract significantly reduced the testosterone-induced proliferation markers, including proliferating cell nuclear antigen and cyclin D1, in the prostate glands of BPH rats; it also reduced levels of androgen receptor, its associated protein steroid 5α-reductase 1 and its downstream target genes (FK506-binding protein 5 and matrix metalloproteinase 2). Notably, compared with the finasteride group, R. vomitoria extract did not significantly reduce sperm count. CONCLUSION: R. vomitoria suppresses testosterone-induced BPH development. Due to its milder side effects, R. vomitoria could be a promising therapeutic agent for BPH.

Modified citrus pectin inhibited bladder tumor growth through downregulation of galectin-3.

Modified citrus pectin (MCP) is a carbohydrate enriched complex, which has been implicated in cancer treatment and prevention. However, the effects of MCP on urinary bladder cancer (UBC) are unknown. In this study, MCP was first tested in T24 and J82 human UBC cells and showed the inhibition of cell viability by the sulforhodamine B (SRB) assay. The MCP-treated UBC cells exhibited G2/M phase arrest with the decrease of Cyclin B1 and phosphorylated Cdc2. Caspase-3 was also activated, leading to the cleavage of Caspase-3 and PARP. We further explored the possible molecular mechanisms upon MCP treatment in UBC cells. Reduction of galectin-3 was observed and followed with the inactivation of Akt signaling pathway. Of note, galectin-3 knockdown by RNA interference recapitulated the MCP-mediated anti-proliferation, cell cycle arrest and apoptosis. Moreover, oral administration of MCP to the T24 xenograft-bearing nude mice inhibited the tumor growth significantly (P < 0.05). Quantification analysis of immunohistochemistry staining for Ki67 and cleaved Caspase-3 confirmed the decrease of proliferation index (P < 0.05) and the increase of apoptosis index (P < 0.01) in 700 mg/kg MCP-fed UBC xenografts. Using the information from TCGA database, we revealed that the overexpression of galectin-3 was associated with high tumor grade with lymph node metastasis, poor overall survival in UBC patients. Considering the remarkable inhibitory effects of MCP on UBC cell proliferation and survival in vitro and in vivo mainly through galectin-3, which is upregulated in UBCs, MCP may become an attractive agent, as a natural dietary fiber, for prevention and therapy of UBCs.

Annexin A5 regulates Leydig cell testosterone production via ERK1/2 pathway.

This study was to investigate the effect of annexin A5 on testosterone secretion from primary rat Leydig cells and the underlying mechanisms. Isolated rat Leydig cells were treated with annexin A5. Testosterone production was detected by chemiluminescence assay. The protein and mRNA of Steroidogenic acute regulatory (StAR), P450scc, 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), and 17α-hydroxylase were examined by Western blotting and semi-quantitative RT-PCR, respectively. Annexin A5 significantly stimulated testosterone secretion from rat Leydig cells in dose- and time-dependent manners and increased mRNA and protein expression of StAR, P450scc, 3ß-HSD, and 17ß-HSD but not 17α-hydroxylase. Annexin A5 knockdown by siRNA significantly decreased the level of testosterone and protein expression of P450scc, 3ß-HSD, and 17ß-HSD. The significant activation of ERK1/2 signaling was observed at 5, 10, and 30 min after annexin A5 treatment. After the pretreatment of Leydig cells with ERK inhibitor PD98059 (50 µmol l-1 ) for 20 min, the effects of annexin A5 on promoting testosterone secretion and increasing the expression of P450scc, 3ß-HSD, and 17ß-HSD were completely abrogated (P < 0.05). Thus, ERK1/2 signaling is involved in the roles of annexin A5 in mediating testosterone production and the expression of P450scc, 3ß-HSD, and 17ß-HSD in Leydig cells.

[Effect of StarD7 and Wnt/ß-catenin signal pathway on the testosterone secretion stimulated by Annexin 5 in rat Leydig cells].

OBJECTIVE: This research aims to study the internal mechanism that promotes the testosterone synthesis by StarD7 and Wnt/ß-catenin, and explores a new regulatory pathway of testosterone synthesis. METHODS: After treated with 1 nmol/L Annexin 5 for 24 h, the culture media were collected for testosterone measurement by chemiluminescence assay. The expressions of StarD7 and ß-catenin at mRNA and protein levels were detected by RT-PCR and western blot respectively. The cellular location of ß-catenin was identified by immunofluorescence. RESULTS: Comparing with the control groups, under the treatment with Annexin 5, the level of testosterone raised 176%[(7.83±0.32)vs.(21.6±1.1), P<0.05], StarD7 mRNA in the experimental groups increased 55%[(1.12±0.08)vs.(1.74±0.11), P<0.05], and ß-catenin mRNA increased 48%[(1.15±0.08)vs.(1.70±0.05), P<0.05]. At the level of protein, the expression of StarD7 in the experimental groups increased 42%[(1.06±0.09)vs.(1.51±0.07), P<0.05], and ß-catenin increased 55%[(1.02± 0.01)vs.(1.58±0.02), P<0.05]. Immunofluorescence identified that ß-catenin was accumulation in the nuclear of the rat Leydig cells in the experiment groups cultured with Annexin 5. CONCLUSION: StarD7 and ß-catenin have both increased significantly at the mRNA and protein levels under treatment with the Annexin 5, and ß-catenin were accumulation in the nuclear of the rat Leydig cells. It suggests that StarD7 and ß-catenin both regulate the effect of Annexin 5 in testosterone production of rat Leydig cells. This regulation may active the Wnt/ß-catenin signal pathway, then increase the expression of the StarD7, eventually raise the progress of the testosterone secretion in rat Leydig cells.

Serum biochemical analysis to indicate pathogenic risk on mouse Mus musculus exposure to source of drinking water.

The 18 biochemical parameters of serum were measured to analyze the pathogenic risks of the Yangtze River Source of Drinking Water in Nanjing area (YZR-SDW-NJ) on mouse Mus musculus for protection of human health in this research. The male mice Mus musculus were sampled and fed with YZR-SDW-NJ for 90 days then the eighteen serum biochemical levels were measured with Automatic Biochemical Analysis/RerLi 600. And the parameter data were treated by One-Way ANOVA statistic approach. The results showed that five parameter levels for the sample group mice were different from those for the control group significantly (0.01 < P or 0.05 < P). Four 4 of the 5 altered parameter levels were decreased including glutamate pyruvate transaminase 38% lower, glutamine-oxaloacetic transaminase 24% lower, triglyceride 76% lower and cystatin C 73% lower, only creatinine level was 26% higher than that in the control group. The data suggest that YZR-SDW-NJ had toxicity on the mouse and the organic pollutants in YZR-SDW-NJ might lead to liver, kidney, cardiovascular and metabolic pathogenic risks on the human beings. The results might be cited as evidence to control pollutants in the source water for the protection of NJ people's health.

[Establishment of colostomy orthotopic transplantation mice and study on their biological characteristics].

OBJECTIVE: To establish a colorectal cancer colostomy orthotopic transplantation mice model. METHODS: A colostomy was preformed in BALB/C nu-nu nude mice. After two weeks, when the stoma healed, tumor tissues developed from Lovo cells were implanted into the submucosa of the stoma. When tumor grew up to 5 mm, fluorouracil(5-FU, 20 mg/kg) was administrated by intraperitoneal injection. Tumor developed at the colostomy was observed and its biological characteristics and behaviour were evaluated. RESULTS: Colostomy was performed in 10 mice and stoma healed at two weeks. Ten colostomies developed detectable tumor in two to three weeks. Three to five weeks later, the tumors grew up to 5 mm. Survival time of mice injected with 5-FU was(15.2+/-3.7) weeks (ranged:11-21 weeks), and the survival time of the no-treatment group was(12.3+/-2.8) weeks(ranged:9-19 weeks). The difference was statistically significant(P=0.001). The rate of mesenteric metastasis was 1/5 and 2/5 in the treatment and no-treatment group respectively. CONCLUSION: Colostomy orthotopic transplantation mice model is an ideal mice model with the advantages of having high success rate, visualization of implanted tumor in living animal, long survival time and significant tumor response to common chemotherapeutic agent.