Panax ginseng C.A. meyer alleviates benign prostatic hyperplasia while preventing finasteride-induced side effects

Abstract

Panax ginseng C.A. Meyer, a widely used traditional medicine in East Asia, shows many beneficial effects on immune function, male erectile dysfunction, cancer, excessive oxidants, and aging issues. However, its effect on benign prostatic hyperplasia (BPH) and its potential in the treatment of side effects related to finasteride (Fi), an FDA-approved drug for BPH, are less known. This study aimed to verify the therapeutic effects of a water extract of P. ginseng (PGWE) on BPH in testosterone propionate (TP)-induced BPH rats and TP-treated RWPE-1 human epithelial cells, and the inhibitory potential on the Fi-induced side effects is also explored. In the TP-induced BPH rat model, PGWE alleviated the pathological markers of BPH such as weight and epithelial thickness of the prostate, and the serum level of dihydrotestosterone. PGWE downregulated androgen-related BPH factors such as 5α-reductase 2 and androgen receptor. PGWE also showed prostatic cell apoptosis accompanied by increased expression of Bax and decreased expression of Bcl-xL and cleaved-caspase 3, respectively, in addition to increasing mitochondrial dynamics in both in vivo and in vitro BPH models. Notably, reduced sperm count, one of the serious side effects of Fi, in the epididymis of BPH rats was recovered with PGWE treatment, suggesting less toxicity to sperm development by PGWE. PGWE also protected against Fi-induced sperm loss when PGWE was administered in combination with Fi without compromising the therapeutic effects of Fi on BPH. Based on these findings, we propose that PGWE could be an alternative therapeutic agent for BPH.

Keywords: 5α-reductase 2; androgen receptor; apoptosis; dihydrotestosterone; sperm loss.

FIGURE 1

LC/MS analysis of PGWE (A) Positive or negative TIC of a cocktail of ginsenoside (Rg1, Rb1, Rg3) was analyzed using the ESI method (B–D) Positive or negative EIC of the PGWE was obtained at m/z 783.5 or at m/z 807.5, m/z 845.5 or m/z 823.5, m/z 1107.6 or m/z 1131.6, respectively. PGWE, Panax ginseng C. A. Meyer water extract; TIC, total ion chromatogram; ESI, electrospray ionization; EIC, extracted ion chromatogram.

FIGURE 2

PGWE alleviates pathological signs of BPH in TP-induced BPH rats (A) Experimental scheme of in vivo study (B) Serum DHT levels were analyzed using ELISA kits (C) Body and (D) prostate weights were assessed (E) Prostate index was calculated by prostate weight (mg)/body weight (100 g) (F) Testis index was calculated by testis weight (mg)/body weight (100 g) (G) H and E staining analysis was performed using prostate tissues of TP-induced BPH rats (magnification × 200, scale bar 150 µm) (H) Based on H and E staining, the epithelial thickness was measured using ImageJ software (J) Number of sperm extracted from the epididymis of rats and (I) length of sperm were calculated using ImageJ software (K, L) Serum levels of AST, ALT, creatinine, and BUN were measured (M) ROS levels were detected by fluorescence microscopy after DCFDA staining in TP-treated RWPE-1 cells. All data are expressed as mean ± SEM of independent experiments. Statistical differences were calculated by one-way ANOVA with post-hoc Tukey’s test. ^# p < 0.05 vs NC groups or D.W.-treated RWPE-1 cells; ^∗ p < 0.05 vs BPH group or TP-treated RWPE-1 cells. NC, normal control; BPH, benign prostatic hyperplasia; TP, testosterone propionate; PGWE, Panax ginseng C. A. Meyer water extract; Fi, finasteride; D.W., distilled water.

FIGURE 3

PGWE reduces 5AR2 and AR levels in prostate tissues of BPH rats and TP-treated RWPE-1 cells (A) The expressions of AR and 5AR2 in prostate tissues of TP-induced BPH rats were analyzed by western blotting. The loading control was β-actin (B) An immunofluorescence assay of AR was conducted in the prostate tissue (magnification × 400, scale bar 75 µm) (C) RWPE-1 cells were treated with various concentrations of PGWE for 24 h to assay cell viability (D, E) Relative mRNA expressions of 5AR2 and AR were determined by RT-PCR (F) The protein expressions of AR, 5AR2, and PSA in RWPE-1 cells were analyzed by western blotting and β-actin was used as the loading control (G) Immunofluorescence assay of AR was conducted in RWPE-1 cells (magnification × 400, scale bar 75 µm). All data are expressed as mean ± SEM of independent experiments. Statistical differences were evaluated using an unpaired t-test and a subsequent post hoc one-tailed Mann-Whitney U test. ^# p < 0.05 vs. NC groups or DW-treated RWPE-1 cells; ^∗ p < 0.05 vs. BPH group or TP-treated RWPE-1 cells. NC, normal control; BPH, benign prostatic hyperplasia; TP, testosterone propionate; PGWE, Panax ginseng C. A. Meyer water extract; Fi, finasteride; D.W., distilled water.

FIGURE 4

PGWE increases DRP1 level in prostate tissue of BPH rats and TP-treated RWPE-1 cells (A) The expressions of MFN1 and DRP1 in prostate tissue of TP-induced BPH rats were analyzed using Western blotting (B) The expressions of MFN1 and DRP1 in TP-treated RWPE-1 cells were analyzed using Western blotting and β-actin was used as the loading control (C) RWPE-1 cells were stained with JC-1 and analyzed with flow cytometry. All data are expressed as mean ± SEM of data from independent experiments. Statistical differences were evaluated using an unpaired t-test and a subsequent post-hoc one-tailed Mann-Whitney U test. ^# p < 0.05 vs NC groups or D.W.-treated RWPE-1 cells; ^∗ p < 0.05 vs BPH group or TP-treated RWPE-1 cells. NC, normal control; BPH, benign prostatic hyperplasia; TP, testosterone propionate; PGWE, Panax ginseng C. A. Meyer water extract; Fi, finasteride; D.W., distilled water.

FIGURE 5

PGWE induces apoptosis in prostate tissue of BPH rats and TP-treated RWPE-1 cells (A) The expressions of Bax, Bcl-xL, caspase-3, and PARP in prostate tissue of TP-induced BPH rats were analyzed using Western blotting (B) The expressions of Bax, Bcl-xL, caspase-3, and PARP in TP-treated RWPE-1 cells were analyzed using Western blotting and β-actin was used as the loading control. All data are expressed as mean ± SEM of independent experiments. Statistical differences were evaluated using an unpaired t-test and a subsequent post-hoc one-tailed Mann-Whitney U test. ^# p < 0.05 vs. NC groups or D.W.-treated RWPE-1 cells; ^∗ p < 0.05 vs. BPH group or TP-treated RWPE-1 cells. NC, normal control; BPH, benign prostatic hyperplasia; TP, testosterone propionate; PGWE, Panax ginseng C. A. Meyer water extract; Fi, finasteride; D. W., distilled water.

FIGURE 6

PGWE restores loss of sperm count by Fi (A) Experimental scheme of in vivo study (B) The prostate index was calculated by prostate weight (mg)/body weight (100 g) (C) The testis index was calculated by testis weight (mg)/body weight (100 g) (D) Serum DHT levels were analyzed using ELISA kits (E) H and E staining analysis was performed using prostate tissues of TP-induced BPH rats (magnification × 200, scale bar 150 µm) (F) Based on H&E staining, the epithelial thickness was measured using ImageJ software (G) Number of sperm extracted from the epididymis of rats and (H) length of sperm were calculated using ImageJ software. All data are expressed as mean ± SEM of independent experiments. Statistical differences were calculated by one-way ANOVA with post-hoc Tukey’s test. ^# p < 0.05 vs. NC group; ^∗ p < 0.05 vs. BPH group. TP, testosterone propionate; NC, normal control; BPH, benign prostatic hyperplasia; PGWE, Panax ginseng C. A. Meyer water extract; Fi, finasteride.