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Proteins were loaded into each lane and resolved by 4% to 12% SDS-polyacrylamide gel electrophoresis. Quantitative proteins (60 µg) were boiled in the loading buffer (62.6 mM Tris-HCl at a pH of 6.8, 2% sodium dodecyl sulfate, 0.01% bromophenol blue, 10% glycerol, and 100 mM dithiothreitol). Protein extracts were quantified with the BCA Protein Assay reagent (Thermo Scientific, Rockford, IL, USA). The bladder total protein was extracted using a cell lysis buffer (20 mM Tris-HCl at a pH of 7.5, 150 mM NaCl, 1 mM Na ethylenediaminetetraacetic acid (EDTA), 1 mM EDTA, 1% Triton, 2.5 mM sodium pyrophosphate, 1 mM β-glycerophosphate, 1 mM Na 3VO 4, 1 µg/mL of leupeptin, and 1 mM phenylmethylsulfonyl fluoride). The plant combination dissolved in distilled water was administered orally through an 8 F red Rob-Nel catheter once a day.įrozen bladder tissue was ground to a fine powder with a liquid nitrogen-cooled mortar and pestle.
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Six of the persistent DO rats were given the plant combination (200 mg/kg) for 4 weeks. The 3-0 nylon was removed through a vaginal approach after 2 weeks.
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The sheath was then removed and 2 ends of nylon were pulled down into the vaginal space through the previously made incision. In the persistent DO and persistent DO with treatment groups, a 25-gauge angioneedle sheath was placed on top of the urethrovesical junction and then ligated with 3-0 nylon to create a partial BOO. An abdominal incision was made to expose the bladder and the urethra. We induced persistent DO according to the method by Jin et al and we confirmed the reliability of the animal model in a preliminary experiment. The 18 rats were randomly assigned to 3 groups: the control group (n=6), the persistent DO group (n=6), and the persistent DO with treatment group (n=6, 200 mg/kg).
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The rats were fed standard rat food and had free access to food and water they were maintained under a 12-hour light-dark cycle, with a room temperature of 20℃☒℃ and a relative humidity of 50%☑0% throughout the experiment. White female Sprague-Dawley rats weighing 250 to 300 g were used in this study. Animal model showing persistent detrusor overactivity after the relief of bladder outlet obstruction Therefore, we studied the effects of the plant combination WSY-1075, which is based on Korean traditional medicine, in animals showing persistent detrusor overactivity (DO) after the relief of BOO.ģ. Recently, several attempts have been made to find candidate herbal treatments showing a treatment effect on BPH and prostate diseases. For these reasons, a new treatment approach is necessary to reduce persistent or de novo storage symptoms after surgery for BPH. Moreover, some patients presented worsening or newly developing storage symptoms after surgery, and their storage symptoms were not improved by medications such as alpha-blockers and antimuscarinics. In particular, previous clinical studies have observed that patients with severe pretreatment storage symptoms experienced persistence and exacerbation of their storage symptoms.
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However, some men with BPH experience persistent or newly developing LUTS after medical and surgical treatment. Medical treatment with various alpha-blockers and surgical procedures such as transurethral resection of the prostate, photoselective vaporization of the prostate, and holmium laser enucleation of the prostate can decrease LUTS by reducing the resistance of the prostatic urethra. Therefore, treatment for BPH aims to reduce BOO and the resistance of the prostatic urethra. Lower urinary tract symptoms (LUTS) in men with BPH are caused by bladder outlet obstruction (BOO) induced by the enlarged prostate gland. Benign prostatic hyperplasia (BPH) is a common type of voiding dysfunction in middle-aged and elderly men.