Protective Effects of KH-204 in the Bladder of Androgen-Deprived Rats

Abstract

Purpose: We investigated the protective effects of the herbal formulation KH-204 in the bladder of androgen-deprived rats.

Materials and methods: Male rats aged eight weeks were randomly divided into four groups, containing eight rats each: sham operation only (normal control group), androgen-deprived only (androgen-deprived control group), and androgen-deprived followed by treatment with 200 mg/kg or 400 mg/kg of KH-204. After 0.5 mg/kg of leuprorelin was subcutaneously injected in the androgen-deprived groups, the oral administration of either distilled water in the two control groups or KH-204 in the treatment group was continued for four weeks. Serum testosterone levels, RhoGEF levels, nitric oxide (NO)-cyclic guanosine monophosphate (cGMP)-related parameters, oxidative stress, and histologic changes were evaluated after treatment.

Results: Treatment with the herbal formulation KH-204 (1) increased serum testosterone levels; (2) restored the expression of RhoGEFs, endothelial NO synthase, and neuronal NO synthase; (3) increased the expression of superoxide dismutase; and (4) decreased bladder fibrosis.

Conclusions: Our results suggest that the positive effects of KH-204 on the urinary bladder may be attributed to its antioxidant effects or to an elevation in NO-cGMP activity.

Keywords: Antioxidants; Hypogonadism; Phytotherapy; Testosterone; Urinary bladder.

Fig. 1. Serum testosterone levels increased by KH-204 treatment. Control: sham operation only, Androgen-dep.: androgen-deprived control group, 200 mg/kg: androgen deprivation followed by treatment with 200 mg/kg of KH-204, 400 mg/kg: androgen-deprivation followed by treatment with 400 mg/kg of KH-204. ^*p<0.05.

Fig. 2. Comparison of the mRNA expression of RhoGEFs in the bladder among the four groups. LARG: leukemia-associated RhoGEF, Control: sham operation only, Androgen dep.: androgen-deprived control group, 200 mg/kg: androgen deprivation followed by treatment with 200 mg/kg of KH-204, 400 mg/kg: androgen-deprivation followed by treatment with 400 mg/kg of KH-204. ^*p<0.05.

Fig. 3. Comparison of the expression levels of eNOS, nNOS and cyclic guanosine monophosphate (cGMP). (A, B) Densitometric analysis of eNOS and nNOS relative to beta-actin and (C) cGMP concentration in bladder tissue. Control: sham operation only, Androgen dep.: androgen-deprived control group, 200 mg/kg: androgen deprivation followed by treatment with 200 mg/kg of KH-204, 400 mg/kg: androgen-deprivation followed by treatment with 400 mg/kg of KH-204, eNOS: endothelial nitric oxide synthase, nNOS: neuronal nitric oxide synthase. ^*p<0.05.

Fig. 4. Comparison of the expression levels of oxdidative stress as reflected by total superoxide dismutase (SOD) expression in bladder tissue. Control: sham operation only, Androgen dep.: androgen-deprived control group, 200 mg/kg: androgen deprivation followed by treatment with 200 mg/kg of KH-204, 400 mg/kg: androgen-deprivation followed by treatment with 400 mg/kg of KH-204. ^*p<0.05.

Fig. 5. Comparison of histologic findings among the four groups. (A) Masson's trichrome staining. The scale bars shown in each figure represent 100 µm. (B) Bladder smooth muscle/collagen ratio. Control: sham operation only, Androgen dep.: androgen androgen-deprived control group, 200 mg/kg: androgen deprivation followed by treatment with 200 mg/kg of KH-204, 400 mg/kg: androgen-deprivation followed by treatment with 400 mg/kg of KH-204. ^*p<0.05.