Effects of Saw Palmetto Extract on Urodynamic Parameters, Bladder Muscarinic and Purinergic Receptors and Urinary Cytokines in Rats with Cyclophosphamide-Induced Cystitis.

OBJECTIVES: To clarify the effect of saw palmetto extract (SPE), a phytotherapeutic agent, on urodynamic parameters, bladder muscarinic and purinergic receptors, and urinary cytokines in rats with cystitis induced by cyclophosphamide (CYP). METHODS: Saw palmetto extract (60 mg/kg per day) was administered orally twice a day for 7 days to rats. The urodynamic parameters in CYP (150 mg/kg i.p.)-treated rats were monitored by a cystometric method under anesthesia. The muscarinic and purinergic receptors in the bladder and submaxillary gland were measured by radioreceptor assays using [N-methyl-(3) H] scopolamine chloride([(3) H]NMS) and αß-methylene-ATP [2,8-(3) H] tetrasodium salt ([(3) H]αß-MeATP), respectively. Urinary cytokines (interleukin-1ß [IL-1ß], IL-6 and L-17) were measured with enzyme linked immunosorbent assay kits. RESULTS: Micturition interval and micturition volume were significantly decreased and the frequency of micturition and basal pressure were significantly increased in the CYP-treated rats compared with sham-operated rats. Orally administered SPE significantly increased the micturition interval and micturition volume and decreased the frequency of micturition and basal pressure. The maximal number of sites (Bmax ) for the specific binding of [(3) H]NMS and [(3) H]αß-MeATP was significantly decreased in the bladder. The decrease in receptors was attenuated by repeated treatment with SPE. An elevation in urinary cytokine (IL-1ß and IL-17) levels were seen, and this increase was effectively suppressed by SPE treatment. CONCLUSIONS: Saw palmetto extract attenuates the alteration of urodynamic parameters, pharmacologically relevant receptors, and urinary cytokines in CYP-treated rats. Therefore, SPE may be a potential therapeutic agent for improving the clinical symptoms of cystitis.

Beneficial effects of Gosha-jinki-gan and green tea extract in rats with chemical cystitis.

The aim of this study was to characterize pharmacological effects of gosha-jinki-gan (GJG) and green tea extract (GTE), on urodynamic parameters, bladder receptors, and urinary cytokines in rats with cyclophosphamide (CYP)-induced cystitis. Urodynamic parameters in CYP-treated rats were measured using the cystometric method. Muscarinic and purinergic receptors in rat tissues were measured by radioreceptor assays. Urinary cytokine levels were measured with ELISA kits. GJG and GTE were orally administered to rats once a day for 7 days. The GJG treatment significantly ameliorated changes in urodynamic parameters in CYP-treated rats. Similar treatment with GTE slightly attenuated changes in urodynamic parameters. The maximal number of binding sites for [³H]NMS and [³H]αß-MeATP in the bladder was significantly lower in CYP-treated rats than in sham rats. Such a reduction in receptor density was significantly attenuated by the GJG treatment. GTE treatment also significantly attenuated the down-regulation of muscarinic receptors, but not P2X receptors in bladders of rats with CYP-induced cystitis. The elevation in urinary cytokine levels in CYP-treated rats was effectively attenuated by GJG treatment. The elevation in cytokine levels in CYP-treated rats was alleviated by GTE treatment. In conclusion, GJG may be a pharmacologically useful plant extract for cystitis.

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.

Characterization of muscarinic receptors in the human bladder mucosa: direct quantification of subtypes using 4-DAMP mustard.

OBJECTIVE: To characterize pharmacologically relevant muscarinic receptors in the human bladder mucosa and detrusor muscle using radioligand binding assays with [N-methyl-3H]scopolamine methyl chloride ([3H]NMS) and 4-DAMP mustard. METHODS: Muscarinic receptors in homogenates of bladder mucosa, detrusor muscle, and parotid gland were measured using the radioligand [3H]NMS. 4-DAMP mustard was used to inactivate M3 receptors irreversibly. RESULTS: Specific [3H]NMS binding in the homogenates of the mucosa and detrusor muscle was saturable and of high affinity as shown by dissociation constants (Kd) of 260 ±82 and 237 ±49 pM, respectively. Antimuscarinic agents (oxybutynin, propiverine, tolterodine, and darifenacin) and their active metabolites competed with [3H]NMS for the binding sites in the human mucosa in a concentration-dependent manner. These agents exhibited similar affinity in the detrusor muscle. The Bmax. values of [3H]NMS in the detrusor, bladder mucosa, and parotid gland were significantly decreased by pretreatment with 4-DAMP mustard (36%, 41% and 63%, respectively). CONCLUSION: The density and binding affinity profile of the muscarinic receptor population in the human bladder mucosa was shown to be similar to that of the detrusor muscle. The density of the M3 subtype in the mucosa was similar to that in the detrusor muscle but lower than that in the parotid gland.