Exploring the potential mechanism of Huang’e capsule (黄莪胶囊) against spontaneous benign prostatic hyperplasia in beagle dogs using high-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry, gas chromatography-mass spectrometry, and network pharmacology

doi:10.19852/j.cnki.jtcm.2026.02.009

Abstract

Abstract:

OBJECTIVE: To investigate the therapeutic efficacy and potential mechanisms of Huang’e capsule (黄莪胶囊, HEC) against benign prostatic hyperplasia (BPH).

METHODS: The chemical profile of HEC was characterized using high-performance liquid chromatographyquadrupole-time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) and gas chromatography-mass spectrometry (GC-MS) techniques. Network pharmacology was employed to analyze potential active compounds, core targets, and key signaling pathways. A spontaneous canine BPH model was used to evaluate the efficacy of HEC and to validate the predictions from network pharmacology.

RESULTS: A total of 51 chemical components of HEC were identified, comprising 19 from HPLC-Q-TOF-MS/MS and 32 from GC-MS analyses. The “components-targets-pathways-disease” network analysis predicted active compounds including (s)-coriolic acid, ethyl linoleate, peroxysimulenoline, physcion, and kaempferol. Core targets identified included cytochrome P450 family 19 subfamily A member 1, estrogen receptor 2 (ESR2), ESR1, and androgen receptor (AR). Kyoto Encyclopedia of Genes and Genomes enrichment analysis suggested that HEC’s effects on BPH involve pathways related to cancer, phosphatidylinositol 3-kinase (PI3K) -protein kinase B (Akt)-signaling, proteoglycans in cancer, and prostate cancer signaling. Animal experiments showed that HEC significantly improved maximum urinary flow rates, reduced prostate weight, volume, and prostate index, and ameliorated histopathological changes. HEC regulated the balance between apoptosis and proliferation by downregulating AR and estrogen receptor alpha expression, while upregulating estrogen receptor beta expression.

CONCLUSION: These findings indicate that HEC effectively ameliorates spontaneous BPH in beagle dogs, likely by regulating the balance between cell apoptosis and proliferation through multiple signaling pathways.

Key words: benign prostatic hyperplasia, proliferation, apoptosis, network pharmacology, Huang’e capsule

MA Aicui, PAN Qi, CHEN Dingshi, WANG Zhonghui, ZHENG Chiyang, ZHANG Junfang, YAN Jianyan, LI Lei, YAN Han, YAO Shunheng, GUO Jun, SUN Zuyue. Exploring the potential mechanism of Huang’e capsule (黄莪胶囊) against spontaneous benign prostatic hyperplasia in beagle dogs using high-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry, gas chromatography-mass spectrometry, and network pharmacology[J]. Journal of Traditional Chinese Medicine, 2026, 46(2): 360-370.

Figures/Tables 6

Table 1 Effects on Qmax and prostate volume by B-mode ultrasonography of HEC pre- and post-administration in beagle dogs with spontaneous BPH ($\stackrel{-}{x}\pm $s)

Group	n	Qmax (mL/s)			Prostate volume by B-scan (cm³)
Group	n	0 week	4 weeks	Difference	0 week	4 weeks	Difference
BPH	7	3.6±1.7	3.6±1.3	-0.0±0.8^a	21.5±2.4	21.7±1.4	0.1±2.6^c
HEC-0.04	7	3.9±1.3	4.6±1.2	0.6±1.3	21.5±3.4	19.3±3.5	-2.2±1.4^c
HEC-0.16	7	3.4±1.5	4.8±2.4	1.4±1.7^b	22.3±3.4	16.8±2.1	-5.5±3.7^d
HEC-0.64	7	3.4±1.4	4.8±2.9	1.4±2.2	21.5±2.9	16.4±1.3	-5.1±3.3^b
HEC-2.56	7	3.3±1.1	4.0±1.4	0.7±1.1	20.9±2.5	14.5±1.7	-6.4±1.8^d
Fina	7	4.8±1.3	6.9±1.8	2.1±1.8^b	20.7±1.6	12.0±1.7	-8.7±1.6^d

Table 2 Effects on prostate volume and prostate weight of HEC in beagle dogs with spontaneous BPH ($\stackrel{-}{x}\pm $s)

Group	n	Prostate volume by water displacement (cm³)	Prostate weight (g)	Prostate index (%)
BPH	7	21.07±4.96^a	22.16±5.49^a	1.48±0.48^a
HEC-0.04	7	20.68±4.54^a	21.17±4.82^a	1.38±0.41^a
HEC-0.16	7	19.90±3.56^a	20.93±3.72^a	1.32±0.25^d
HEC-0.64	7	18.16±3.65^a	18.45±3.69^d	1.34±0.31^d
HEC-2.56	7	14.13±1.08^b	14.76±0.91^b	1.05±0.13^b
Fina	7	10.85±2.80^c	11.41±2.99^c	0.74±0.18^c

Figure 1 Effect of HEC in prostate tissues in beagle dogs with spontaneous BPH A: histopathological study of prostate tissues in beagle dogs with spontaneous BPH using HE staining; A1: BPH group (× 50, scale bar = 500 μm); A2: BPH group (× 200, scale bar = 100 μm); A3: HEC-0.04 group (× 50, scale bar = 500 μm); A4: HEC-0.04 group (× 200, scale bar = 100 μm); A5: HEC-0.16 group (× 50, scale bar = 500 μm); A6: HEC-0.16 group (× 200, scale bar = 100 μm); A7: HEC-0.64 group (× 50, scale bar = 500 μm); A8: HEC-0.64 group (× 200, scale bar = 100 μm); A9: HEC-2.56 group (× 50, scale bar = 500 μm); A10: HEC-2.56 group (× 200, scale bar = 100 μm); A11: Fina group (× 50, scale bar = 500 μm); A12: Fina group (× 200, scale bar = 100 μm); B: the epithelial thickness of the prostates in beagle dogs with spontaneous BPH; C: the lumen area of the prostates in beagle dogs with spontaneous BPH. BPH: the BPH group; HEC-0.04, HEC-0.16, HEC-0.64, HEC-2.56: 0.04, 0.16, 0.64, 2.56 g/kg HEC-treated group, respectively; Fina: 0.24 mg/kg finasteride-treated group. HE: hematoxylin and eosin; HEC: Huang’e capsule; BPH: benign prostatic hyperplasia. Differences among three or more groups were analyzed by analysis of variance. Data are presented as mean ± standard deviation (n = 7 per group). aP   0.01, bP   0.05, compared to Fina group; cP   0.01, dP   0.05, compared to BPH group.

Figure 2 Effect of HEC on the expression of AR, ERα, and ERβ in prostate tissues in beagle dogs with spontaneous BPH A: slices of prostate tissue from each group were examined for AR using IHC (scale bar = 100 μm); A1: BPH group; A2: HEC-0.04 group; A3: HEC-0.16 group; A4: HEC-0.64 group; A5: HEC-2.56 group; A6: Fina group; B: the proportion of AR-positive cells in prostatic cells was estimated; C: slices of prostate tissue from each group were examined for ERα using IHC (scale bar = 100 μm); C1: BPH group; C2: HEC-0.04 group; C3: HEC-0.16 group; C4: HEC-0.64 group; C5: HEC-2.56 group; C6: Fina group; D: the proportion of cells that were ERα positive was calculated; E: slices of prostate tissue from each group were examined for ERβ using IHC (scale bar = 100 μm); E1: BPH group; E2: HEC-0.04 group; E3: HEC-0.16 group; E4: HEC-0.64 group; E5: HEC-2.56 group; E6: Fina group; F: the proportion of cells that were ERβ positive was calculated. BPH: the BPH group; HEC-0.04, HEC-0.16, HEC-0.64, HEC-2.56: 0.04, 0.16, 0.64, 2.56 g/kg HEC-treated group, respectively; Fina: 0.24 mg/kg finasteride-treated group. HEC: Huang’e capsule; BPH: benign prostatic hyperplasia; AR: androgen receptor; ERα: estrogen receptor alpha, ERβ: estrogen receptor beta; IHC: immunohistochemical. Differences among three or more groups were analyzed by analysis of variance. Data are presented as mean ± standard deviation (n = 7 per group). aP   0.05, compared to Fina group; bP   0.05, compared to BPH group.

Figure 3 Effect of HEC on prostatic cell proliferation in beagle dogs with spontaneous BPH A: the localization of PCNA in each group was analyzed by IHC tests (scale bar = 50 μm); A1: BPH group; A2: HEC-0.04 group; A3: HEC-0.16 group; A4: HEC-0.64 group; A5: HEC-2.56 group; A6: Fina group; B: the proportion of PCNA-positive cells percentages was displayed. BPH: the BPH group; HEC-0.04, HEC-0.16, HEC-0.64, HEC-2.56: 0.04, 0.16, 0.64, 2.56 g/kg HEC-treated group, respectively; Fina: 0.24 mg/kg finasteride-treated group. PCNA: proliferating cell nuclear antigen; HEC: Huang’e capsule; BPH: benign prostatic hyperplasia. Differences among three or more groups were analyzed by analysis of variance. Data are presented as mean ± standard deviation (n = 7 per group). aP   0.05, compared to Fina group; bP   0.05, cP   0.01, compared to BPH group.

Figure 4 Effect of HEC on prostatic cell apoptosis in beagle dogs with spontaneous BPH A: the localization of TUNEL-positive cells in each group (scale bar = 50 μm); A1: BPH group; A2: HEC-0.04 group; A3: HEC-0.16 group; A4: HEC-0.64 group; A5: HEC-2.56 group; A6: Fina group; B: the proportion of TUNEL-positive cells was displayed. BPH: the BPH group; HEC-0.04, HEC-0.16, HEC-0.64, HEC-2.56: 0.04, 0.16, 0.64, 2.56 g/kg HEC-treated group, respectively; Fina: 0.24 mg/kg finasteride-treated group. TUNEL: terminal deoxynucleotidyl transferase dUTP nick end labeling; HEC: Huang’e capsule; BPH: benign prostatic hyperplasia. Differences among three or more groups were analyzed by analysis of variance. Data are presented as mean ± standard deviation (n = 7 per group). aP   0.05, compared to Fina group; bP   0.01, cP   0.05, compared to BPH group.

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