The phytotherapeutic agent, eviprostat, suppresses stromal proliferation and inflammation even after establishment of nonbacterial prostatitis in the rat prostate.

OBJECTIVE: To evaluate the effect of phytotherapeutic agent, Eviprostat, administered after the establishment of nonbacterial prostatitis (NBP) on the stroma-to-epithelium ratio (S/E ratio), inflammatory scores, tissue macrophage infiltration, and cytokines and chemokines levels in prostate tissue and urine. MATERIALS AND METHODS: Ten-month-old male Wistar rats were castrated and exposed to 17-beta-isomer of estradiol for 30 days to induce NBP. Twenty-five NBP rats were divided into 5 groups: (1) NBP (0) rats sacrificed immediately after the establishment of NBP; (2,3) NBP (30)/control (CTL) and NBP (30)/Eviprostat (EVI) rats fed without or with 0.1% Eviprostat under estradiol-free for 30 days, respectively; and (4,5) NBP (60)/CTL and NBP (60)/EVI rats fed without or with 0.1% Eviprostat under estradiol-free for 60 days, respectively. The S/E ratio, inflammatory scores, and the number of macrophage infiltration in the prostate were assessed. Concentrations of cytokines and chemokines in prostatic tissue and urine were measured by enzyme-linked immunosorbent assay. RESULTS: The S/E ratio was significantly increased with time until 60 days under estradiol-free condition (P <.001). The S/E ratio and the inflammatory scores in NBP (60)/EVI was significantly lower than that of NBP (60)/CTL (P <.001, and P = .022, respectively). The mean tissue concentration of chemokine ligand 2 (CCL2)/monocyte chemoattractant protein-1 (MCP-1) in NBP (60)/CTL was significantly higher than that in NBP (0) (P = .016), whereas, there was no difference between NBP (60)/EVI and NBP (0). Furthermore, urinary CCL2/MCP-1 was significantly decreased in NBP (60)/EVI as compared with NBP (0) (P = .028). CONCLUSION: Eviprostat suppresses the stromal proliferation and inflammation in the rat prostate after the establishment of NBP at least partly owing to inhibitory effect on CCL2/MCP-1 production in the prostate.

Eviprostat activates cAMP signaling pathway and suppresses bladder smooth muscle cell proliferation.

Eviprostat is a popular phytotherapeutic agent for the treatment of lower urinary tract symptoms (LUTS). At present, the signaling mechanisms underlying its therapeutic effects are still poorly understood. Given that cAMP has been reported to suppress cell hyperplasia and hypertrophy in various pathological situations, we asked whether the effect of Eviprostat could be ascribed to the activation of the cAMP signaling pathway. In the study, exposure of cAMP response element (CRE)-secreted alkaline phosphatase (SEAP) (CRE-SEAP)-reporter cells to Eviprostat elevated SEAP secretion, which was associated with an increased phosphorylation of vasodilator-stimulated phosphoprotein (VASP) and cAMP-response element-binding protein (CREB), as well as enhanced expression of CRE-regulated protein connexin43, indicating an activation of the cAMP signaling pathway. Consistent with these observations, Eviprostat-induced expression of Cx43 was abolished in the presence of adenylyl cyclase inhibitor SQ22536 or PKA inhibitor H89, whereas it was mimicked by adenylyl cyclase activator, forskolin. Further analysis demonstrated that Eviprostat significantly potentiated the effect of phosphodiesterase 3 (PDE3) inhibitor, but not that of PDE4 inhibitor, on CRE activation. Moreover, Eviprostat suppressed PDGF-induced activation of ERK and Akt and inhibited cell proliferation and hillock formation in both mesangial cells and bladder smooth muscle cells. Collectively, activation of the cAMP signaling pathway could be an important mechanism by which Eviprostat exerts its therapeutic effects for LUTS.

Improvement by phytotherapeutic agent of detrusor overactivity, down-regulation of pharmacological receptors and urinary cytokines in rats with cyclophosphamide induced cystitis.

PURPOSE: We characterized pharmacological effects of the phytotherapeutic agent Eviprostat® on urodynamic parameters, bladder muscarinic and purinergic receptors, and urinary cytokines in rats with cyclophosphamide induced cystitis. MATERIALS AND METHODS: Urodynamic parameters in cyclophosphamide (150 mg/kg intraperitoneally) treated rats were measured by a cystometric method. Muscarinic and purinergic receptors in the bladder and other tissues were measured by radioreceptor assays using [N-methyl-(3)H]scopolamine methyl chloride and [(3)H]αß-MeATP, respectively. The urinary cytokines interleukin-1ß, 6 and 17 were measured with enzyme-linked immunoassay kits. Eviprostat (36 mg/kg per day twice daily for 7 days) was orally administered. RESULTS: On cystometry the micturition interval and micturition volume were significantly decreased in cyclophosphamide vs sham treated rats, while micturition frequency, basal pressure and post-void residual urine volume were significantly increased. Repeat oral administration of Eviprostat in cyclophosphamide treated rats significantly increased the micturition interval and micturition volume, and decreased micturition frequency, basal pressure and post-void residual urine volume. The maximal number of binding sites for [N-methyl-(3)H]scopolamine methyl chloride and [(3)H]αß-MeATP was significantly decreased in the bladder of cyclophosphamide vs sham treated rats. Such decreases were significantly attenuated by repeat Eviprostat treatment. Increased urinary cytokine levels in cyclophosphamide treated rats were also effectively attenuated by Eviprostat. CONCLUSIONS: Repeat Eviprostat treatment significantly improved detrusor overactivity, down-regulated the expression of bladder pharmacological receptors and increased urinary cytokine levels in rats with cyclophosphamide induced cystitis. Therefore, Eviprostat may be a pharmacologically useful phytotherapeutic agent for cystitis.

Effect of the phytotherapeutic agent eviprostat on the bladder in a rat model of bladder overdistension/emptying.

AIMS: Ischemia-reperfusion injury is an important factor in the development of lower urinary tract symptoms (LUTS) that is partly mediated by the generation of free radicals. We investigate the effect of the phytotherapeutic agent Eviprostat, a treatment for benign prostatic hyperplasia (BPH) that has antioxidant and anti-inflammatory activity, on urinary bladder blood flow (BBF), and function in a rat model of bladder overdistension and emptying (OE). METHODS: For 8 days before surgery, OE rats received daily oral Eviprostat (36 mg/kg/day) or vehicle, while sham-operated animals received vehicle. The bladder was distended by infusion of saline over a period of 2 hr (overdistension) and then emptied. After 24 hr, BBF was measured with a laser speckle blood flow imager. The oxidative-stress marker malondialdehyde (MDA), proinflammatory cytokines, and myeloperoxidase were determined in the isolated bladder, and histological analysis was performed. Functional contractile responses of bladder strips to electrical field stimulation, carbachol, and KCl were measured. RESULTS: Twenty-four hours after bladder OE, a significant decrease in BBF and significant increases in bladder weight, malondialdehyde, proinflammatory cytokines, and myeloperoxidase were observed. Eviprostat almost completely prevented these changes. Histological analysis of the bladder of OE rats showed hemorrhage, accumulation of leukocytes, desquamation of epithelium, and edema, and Eviprostat suppressed these changes. The reduction in functional contractile forces in the bladder of OE rats was also prevented by Eviprostat. CONCLUSION: Eviprostat-mediated suppression of increased bladder oxidative stress and inflammation caused by bladder OE may contribute to the improvement of BBF and bladder function by Eviprostat.

Suppression of bladder overactivity and oxidative stress by the phytotherapeutic agent, Eviprostat, in a rat model of atherosclerosis-induced chronic bladder ischemia.

OBJECTIVES: To clarify the mechanism by which chronic bladder ischemia causes bladder functional changes, and to investigate the involvement of oxidative stress and pro-inflammatory cytokines, and the effects of the phytotherapeutic drug, Eviprostat, on these biochemical marker levels and bladder function. METHODS: Male Sprague-Dawley rats aged 15 weeks were divided into three groups. Arterial injury was experimentally induced by balloon endothelial injury of the iliac arteries, and a 2% cholesterol diet was given for 8 weeks. Rats in the arterial-injury group were given daily oral vehicle or Eviprostat, whereas sham-operated animals on a regular diet (0.09% cholesterol) were given vehicle for the last 2 weeks. Eight weeks after surgery, the levels of bladder pro-inflammatory cytokines, as well as bladder and urinary oxidative-stress markers, were determined. Cystometrograms were carried out without anesthesia or restraint. RESULTS: Bladder and urinary oxidative-stress markers, and bladder pro-inflammatory cytokine levels were significantly increased in the arterial-injury group, and Eviprostat markedly suppressed these increase. The cystometrograms showed that arterial injury decreased the intermicturition interval without affecting the micturition pressure. This decrease was reversed by Eviprostat treatment. CONCLUSIONS: Oxidative stress and pro-inflammatory cytokines might be involved in the development of overactive bladder by atherosclerosis-induced chronic bladder ischemia. Eviprostat might provide an attractive treatment option for individuals with bladder dysfunction due to chronic bladder ischemia because of its anti-oxidant and anti-inflammatory properties.

Effect of the phytotherapeutic agent Eviprostat on inflammatory changes and cytokine production in a rat model of nonbacterial prostatitis.

OBJECTIVES: To examine the effect of the phytotherapeutic agent Eviprostat on the stromal-to-epithelial (S/E) ratio, level of macrophage infiltration, expression of the macrophage inhibitory cytokine-1 (Mic1) gene, and tumor necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8) concentrations in prostate tissues in a rat model of nonbacterial prostatitis (NBP). MATERIALS AND METHODS: Ten-month old Wistar rats were divided into 4 groups of 10: (1) NBP non-mixed feed (prostatitis control group); (2) NBP Eviprostat (0.1%) mixed feed (prostatitis Eviprostat group); (3) non-NBP non-mixed feed (nonprostatitis control group); and (4) non-NBP Eviprostat mixed-feed (nonprostatitis Eviprostat group). NBP was induced by castration followed by daily subcutaneous injection of 17ß-estradiol for 30 days. Ventral prostate lobes were histopathologically examined with Masson's trichrome staining or immunostaining with antimacrophage antibody. Mic1 mRNA levels were quantified by real-time reverse transcriptase polymerase chain reaction. Tissue concentrations of TNF-α and IL-8 were determined by enzyme-linked immunosorbent assay. RESULTS: Stroma was the most abundant in prostatitis control rats. The mean S/E ratio in prostatitis Eviprostat rats was significantly lower than in prostatitis control rats (P < .0001). The high levels of macrophage infiltration found in prostatitis control rats were significantly reduced in prostatitis Eviprostat rats (P < .0001). The up-regulation of the Mic1 gene observed in prostatitis control rat prostates was significantly suppressed in prostatitis Eviprostat rats (P < .0001). A marked suppression of TNF-α and IL-8 secretion was also observed in prostatitis Eviprostat rats (P < .05). CONCLUSIONS: Eviprostat significantly suppressed the S/E ratio, level of macrophage infiltration, Mic1 gene expression, and proinflammatory cytokines/chemokines in the prostate in a rat NBP model.

Effect of a phytotherapeutic agent, Eviprostat®, on prostatic and urinary cytokines/chemokines in a rat model of nonbacterial prostatitis.

BACKGROUND: Chronic inflammation in the prostate has recently been recognized as an important component of the symptom progression of benign prostatic hyperplasia. The objective of this study was to evaluate a range of cytokines/chemokines in prostate tissue and urine to identify markers of prostate inflammation in a prostatitis model and to investigate the effect of a phytotherapeutic agent, Eviprostat®, on these markers. METHODS: Ten-month-old male Wistar rats were divided into four groups. Nonbacterial prostatitis (NBP) was experimentally induced in groups 2-4 by castration followed by daily subcutaneous injection of 17ß-estradiol for 30 days. Control rats were fed a standard diet, while animals in the Eviprostat groups were fed a diet containing 0.05 or 0.1% Eviprostat for 30 days. The levels of cytokines/chemokines in prostate tissue on the 31st day and in urine collected the day before castration and the day before removal of the prostate were determined. RESULTS: Experimentally induced NBP increased the prostatic levels of the cytokines interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α). The levels of the chemokines CCL2/monocyte chemoattractant protein-1 (MCP-1), CCL3/macrophage inflammatory protein-1α (MIP-1α), CXCL1/CINC-1, CXCL3/CINC-2, and CXCL5/LIX were elevated in both prostate and urine. Eviprostat significantly suppressed the increases in prostate IL-1ß, TNF-α and CCL3/MIP-1α and prostatic and urinary CCL2/MCP-1 and CXCL1/CINC-1. CONCLUSIONS: Chemokines, including CCL2/MCP-1 and CXCL1/CINC-1, were elevated in the prostate and urine of NBP rats, and Eviprostat potently suppressed the increases in CCL2/MCP-1 and CXCL1/CINC-1. These chemokines are therefore candidate diagnostic biomarkers for nonbacterial chronic prostatic inflammation.

Eviprostat suppresses proinflammatory gene expression in the prostate of rats with partial bladder-outlet obstruction: a genome-wide DNA microarray analysis.

Prostatic inflammation plays a role in the progression of benign prostatic hyperplasia (BPH). Eviprostat is an antioxidant, antiinflammatory phytotherapeutic agent widely used to treat lower urinary tract symptoms in BPH. Because Eviprostat is a mixture of compounds from multiple natural sources, however, its mechanism of action has been difficult to investigate. Here, we describe the use of oligonucleotide microarrays to investigate changes in gene expression in the prostate of rats with surgically induced partial bladder-outlet obstruction and the effect of Eviprostat on those changes. Several dozen proinflammatory genes were activated in obstructed rats, including cytokine, arachidonic acid cascade enzyme, Toll-like receptor (TLR), and transcription factor genes, and their expression was suppressed by Eviprostat. Pathway analysis revealed that several proinflammatory pathways were activated, including cytokine and TLR signaling pathways. The differential expression of selected genes was verified by real-time reverse-transcriptase polymerase chain reaction. Our findings suggest that prostate inflammation in our rat model of partial bladder-outlet obstruction is related to the increased expression of nuclear factor kappaB (NF-kappaB) and the induction of proinflammatory cytokines, and that Eviprostat suppresses their expression at the transcriptional level. The prostate inflammation seen in BPH and the clinical benefits of Eviprostat may be similarly explained.

Effect of the phytotherapeutic agent Eviprostat on 17beta-estradiol-induced nonbacterial inflammation in the rat prostate.

BACKGROUND: Anti-inflammatory medications have been used for the treatment of chronic prostatitis. The phytotherapeutic agent Eviprostat, a popular treatment for benign prostatic hyperplasia in Japan and Germany, has antioxidant and anti-inflammatory activity. We investigated the effects of the phytotherapeutic agent Eviprostat on prostate inflammation induced in castrated rats by the injection of 17beta-estradiol. METHODS: Ten-month-old male Wistar rats were divided into five groups. Nonbacterial prostatitis was experimentally induced in groups 2-5 by castration followed by daily subcutaneous injection of 17beta-estradiol for 30 days. The rats were orally administered 0.1% Tween-80 (group 2), low-dose Eviprostat (group 3), high-dose Eviprostat (group 4), or cernitin pollen extract (group 5) for the last 2 weeks of 17beta-estradiol administration. Sham-operated rats (group 1) were orally administered 0.1% Tween-80. On the 31st day after surgery, the weight of the prostate and the levels of prostatic proinflammatory cytokines as well as the oxidative-stress marker malondialdehyde were determined and histological alterations noted. RESULTS: Experimentally induced nonbacterial prostatitis led to a significant decrease in prostate weight and increases in malondialdehyde and proinflammatory cytokine levels. Eviprostat significantly suppressed the increases in malondialdehyde and cytokine levels without affecting prostate weight. Histologically, nonbacterial prostatitis was evident in the lateral lobe of the prostate, and Eviprostat treatment significantly suppressed the severity of the lesion. CONCLUSIONS: Eviprostat, which has effective antioxidant and anti-inflammatory activities in the prostate, may be useful for the clinical treatment of chronic prostatitis. These activities of Eviprostat may also contribute to the amelioration of prostate inflammation in BPH patients. Prostate 69: 1404-1410, 2009. (c) 2009 Wiley-Liss, Inc.

Suppression of bladder oxidative stress and inflammation by a phytotherapeutic agent in a rat model of partial bladder outlet obstruction.

PURPOSE: Ischemia/reperfusion injury is a major etiological factor in the progression of bladder dysfunction after partial bladder outlet obstruction and it is partly mediated by the generation of free radicals. The phytotherapeutic agent Eviprostat, a popular treatment for benign prostatic hyperplasia in Japan and Germany, has antioxidant and anti-inflammatory activity. We investigated the effect of Eviprostat on oxidative stress and inflammation in bladder dysfunction in a bladder outlet obstruction rat model. MATERIALS AND METHODS: Bladder outlet obstruction was surgically induced in male rats by placing a rubber ring around the urethra. Rats with bladder outlet obstruction were administered daily oral Eviprostat or vehicle, while sham operated animals were treated with vehicle. On day 6 after surgery bladder weight, oxidative stress markers and proinflammatory cytokine levels as a measure of bladder inflammation, were determined and histological alterations noted. Functional contractility studies were performed with longitudinal bladder strips. RESULTS: Bladder outlet obstruction led to a significant increase in bladder weight, oxidative stress markers and proinflammatory cytokine levels. Eviprostat significantly suppressed these increases without affecting bladder weight. Histological analysis showed increased detrusor muscle hypertrophy and increased numbers of collagen fibers with accompanying inflammatory infiltration in the bladder of vehicle treated bladder outlet obstruction animals. Eviprostat treatment was associated with suppression of these changes. Decreased responses of obstructed bladder strips to electrical stimulation and KCl were ameliorated by Eviprostat treatment. CONCLUSIONS: Eviprostat mediated decrease of the increased oxidative stress and bladder inflammation caused by bladder outlet obstruction may contribute to the protection of bladder function.

Effect of eviprostat on bladder overactivity in an experimental cystitis rat model.

OBJECTIVE: The purpose of this study was to investigate the effects of eviprostat, a phytotherapeutic drug, on bladder overactivity and inflammation in a cyclophosphamide (CYP)-induced cystitis rat model. METHODS: Female Sprague-Dawley rats received a single intraperitoneal injection of CYP (200 mg/kg) or saline. After the CYP injection, eviprostat (9, 18 or 54 mg/kg per day) or a vehicle was orally given twice each day. Four days after the CYP injection, bladder function was evaluated by cystometrograms under urethane anesthesia. In a separate group, bladder inflammation was compared between the eviprostat- or vehicle-treated animals. Furthermore, the effects of eviprostat on carbachol-induced muscle contraction were evaluated by an in vitro experiment. RESULTS: The intercontraction interval (ICI) significantly decreased in the CYP-injected rats in comparison to the saline-injected rats. In the CYP-injected group, 18 and 54 mg/kg per day of eviprostat treatment significantly increased the ICI, but did not change the maximum voiding pressure in comparison to the vehicle treatment. In the saline-injected group, no significant changes of any parameters in the cystometrograms were observed between the eviprostat- and vehicle-treated groups. CYP-induced bladder inflammation tended to be lower in the eviprostat-treated group in comparison to the vehicle-treated group. An in vitro experiment revealed that eviprostat failed to inhibit carbachol-induced muscle contraction. CONCLUSION: The oral administration of eviprostat suppressed CYP-induced bladder overactivity. The effects of eviprostat on the micturition reflex may be irrespective of antimuscarinic action. The present findings raise the possibility that eviprostat could be an effective treatment for bladder overactivity associated with inflammation.

Mechanisms by which a phytotherapeutic drug influences bladder activity in rats.

PURPOSE: We investigated the mechanisms by which Eviprostat, a phytotherapeutic drug for benign prostatic hyperplasia, influences bladder activity in rats. MATERIALS AND METHODS: A total of 42 female rats were divided into a control group and an Eviprostat group. Rats in the control group were fed a standard diet, while animals in the Eviprostat group were fed a diet containing 0.1% Eviprostat. After 2 weeks 14 rats (7 rats per group) underwent continuous cystometry with physiological saline or 0.1% acetic acid solution and bladder activity was recorded. Body weight, blood pressure, plasma monoamines and adenosine triphosphate were measured in another 14 rats (7 per group). In the remaining 14 rats (7 per group) 0.1% acetic acid solution was infused into the bladder and urinary adenosine triphosphate was measured before and after stimulation. RESULTS: During cystometry with acetic acid the interval between bladder contractions was shorter and maximum bladder contraction pressure was higher in the control group compared with results obtained using physiological saline but such differences were not seen in the Eviprostat group. Plasma adrenaline and noradrenaline were lower in the Eviprostat group than the control group but no difference in blood pressure was observed. Urinary adenosine triphosphate was higher in the 2 groups than before stimulation but the increase was smaller in the Eviprostat group than in the control group. CONCLUSIONS: These results suggest that Eviprostat acts to maintain low catecholamine and also inhibit pathological bladder activity by decreasing adenosine triphosphate release from the bladder.

Relevance of anti-reactive oxygen species activity to anti-inflammatory activity of components of eviprostat, a phytotherapeutic agent for benign prostatic hyperplasia.

Inflammation is a common finding in benign prostatic hyperplasia (BPH). The phytotherapeutic agent eviprostat is a popular treatment for BPH in Japan and Germany. This agent consists of five components; four are extracted from Chimaphila umbellata, Populus tremula, Pulsatilla pratensis and Equisetum arvense (coded as EVI-1, EVI-2, EVI-3 and EVI-4, respectively) and the fifth is germ oil from Triticum aestivum (coded as EVI-5). In this study, the effects of each component on the reactive oxygen species (ROS), superoxide anion (O2-) and hydroxyl radical (OH*) generated in cell-free systems and human neutrophils, and on carrageenin-induced paw edema in rats were investigated. EVI-1, EVI-2 and EVI-4 suppressed the O2- levels in the xanthine/xanthine oxidase system, and EVI-1, EVI-2, EVI-3 and EVI-4 abolished the OH* produced in a Fenton-type reaction system, so that EVI-1, EVI-2 and EVI-4 possessed inhibitory action with respect to both O2- and OH*. EVI-1, EVI-2 and EVI-4 also reduced ROS levels in phorbol myristate acetate-stimulated neutrophils. The paw swelling was inhibited by a mixture of EVI-1, EVI-2, EVI-3, EVI-4 and EVI-5 (a mixture which is equivalent to eviprostat) or by a mixture of EVI-1, EVI-2 and EVI-4, even though each component alone did not significantly inhibit the swelling. These findings suggest that the suppression of ROS by EVI-1, EVI-2 and EVI-4 may partly contribute to the anti-inflammatory action of eviprostat, and this action may be implicated in its therapeutic effect on BPH.

Actualización del tratamiento médico de la hemorragiauterina disfuncional / An update of the medical management of dysfunctional uterine bleeding

La hemorragia uterina disfuncional es un transtorno frecuente ginecológico. Es un diagnóstico de exclusión, y el clínico debe proceder a una evaluación lógica y escalonada de todas las causas de sangrado anormal. La menorragia en la mayoría de los casos se asocia con anovulación. El tratamiento médico debería ser la primera opción terapéutica y puede ser dividido en terapia no hormonal y hormonal. Los dos principales tratamientos para la menorragia asociada a ciclos ovulatorios son no hormonales, mediante un antifibrinolítico como el ácido tranexámico y antiinflamatorios. Tradicionalmente la terapia hormonal para la menorragia ha estado constituida por los progestágenos y los anticonceptivos orales. El sistema intrauterino de liberación de levonorgestrel ofrece un nuevo concepto terapéutico que combina una eficaz contracepción con una reducción del sangrado menstrual. Es una buena alternativa conservadora a la resección endometrial y parece ser una importante alternativa a la medicación oral. En el manejo de la pérdida menstrual excesiva hay una evidencia demostrada de que muchos médicos no prescriben los tratamientos más adecuados. El incremento en la utilización de tratamientos efectivos mejoraría las expectativas de las pacientes y supondría una alternativa a la cirugía (AU)