Study On The Mechanism Of Tongshuang Prescription For Benign Prostatic Hyperplasia Based On UPLC‑Q‑TOF/MS And Integrated Network Pharmacology
Jan 05, 2026
Why this matters: Lower urinary tract symptoms (LUTS) from benign prostatic hyperplasia (BPH) affect quality of life. While α‑blockers and 5‑ARI drugs work, many men seek botanicals to reduce nocturia, frequency, and hesitancy with fewer sexual or blood-pressure side effects.
Where herbs may fit: Standardized, mechanism‑guided herbal actives can complement lifestyle and conventional care. Classic formulas like Tongshuang (Tongshuang Fang) and modern adaptogens such as Cistanche spp. have multi‑target anti‑inflammatory and hormone‑modulating actions that align with BPH biology and chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS).
Related reading: Overview on Cistanche for prostate health (plain‑language explainer) https://www.xjcistanche.com/news/say-goodbye-to-prostate-problems-with-cistanch-83534746.html
HERB CISTANCHE SUPPLEMENT BENEFITS
Abstract
Objective: To identify the active constituents and potential mechanisms of Tongshuang Prescription (TS) in treating benign prostatic hyperplasia (BPH).
Methods: Ultra‑performance liquid chromatography–quadrupole time‑of‑flight mass spectrometry (UPLC‑Q‑TOF/MS) characterized TS components. Putative targets of TS compounds were predicted via SwissTargetPrediction; BPH‑related targets were gathered from GeneCards and OMIM. Common targets were used to construct a high‑confidence protein–protein interaction (PPI) network on STRING. GO and KEGG enrichment analyses were performed with Metascape. A "drug–component–target–disease" network was built in Cytoscape 3.9.1. Key compounds were docked to core targets using AutoDock; structures were visualized in PyMOL.
Results: Thirty‑six compounds were identified. Network pharmacology yielded 713 TS component targets, 1,667 disease targets, and 218 intersecting targets. Five core targets emerged: GAPDH, TP53, TNF, PTGS2 (COX‑2), and ESR1. Molecular docking indicated that daidzein, styrene, β‑ionone, kaempferol, quercetin, amaranth sterone, m‑cymene, myricetin, formononetin, and 7‑methoxycoumarin showed favorable binding to these core targets.
Conclusion: TS likely acts through multi‑component regulation of inflammation and hormone signaling-particularly TNF and COX‑2 pathways and estrogen receptor signaling-helping relieve BPH‑related LUTS. Flavonoids and isoflavones (e.g., quercetin, kaempferol, daidzein, formononetin) appear to be principal bioactives. This supports TS as a candidate for standardized development and clinical evaluation.
Keywords
Benign prostatic hyperplasia; Tongshuang Prescription; UPLC‑Q‑TOF/MS; network pharmacology; molecular docking; TNF; PTGS2/COX‑2; ESR1; quercetin; daidzein; Cistanche; What is the best herbal for prostate?
Supportive Service Of Wecistanche-For more details about cooperation
Email:wallence.suen@wecistanche.com
Background and formula rationale
Tongshuang Prescription derives from "Lougu San" in Puji Fang and is a five‑herb classic modified by experienced TCM clinicians: Gryllus bimaculatus (Mole cricket), Salty Semen Astragali Complanati (Cuscuta-like "Sha Yuan Zi" processed with salt), Achyranthes bidentata, Lygodium japonicum spores (Hai Jin Sha), and Cinnamomum cassia (Rou Gui).
Sovereign actions: Gryllus to strongly promote urination; salt‑processed Sha Yuan Zi to tonify kidney qi and secure essence, reducing nocturia and dribbling.
Minister: Achyranthes to tonify liver–kidney and promote diuresis.
Assistant: Lygodium spores to clear heat, relieve stranguria.
Courier: Cinnamon bark to warm the gate of vitality and support qi transformation for smooth urination.
Clinically, TS has been used to alleviate LUTS in BPH with good tolerability; however, active ingredients and mechanisms remained unclear, motivating this study.
Methods in brief
Instruments: Shimadzu LC‑30AD UPLC; AB Sciex TripleTOF 5600 MS; PeakView 1.2; Analyst TF 1.6; Cytoscape 3.9.1; AutoDock; PyMOL.
Chemicals and reference standards: authenticated herbal decoction pieces; standards including astragaloside A, amaranth sterone, cinnamic acid, cinnamaldehyde (all ≥99%).
Sample prep: TS decoction prepared via two-stage water extraction; combined filtrates concentrated to approximately 1.0 g raw herb/mL.
UPLC conditions: ACQUITY UPLC BEH C18, 100×2.1 mm, 1.7 μm; acetonitrile–0.1% phosphoric acid gradient; 0.3 mL/min; 40°C; 2 μL injection.
MS conditions: ESI positive/negative; ISVF −4500 V; temp 500°C; CUR 30 psi; GS1/GS2 55 psi; DP −100 V; CE 30 eV with ±15 eV ramp; m/z 80–1250.
Target mining and enrichment: SwissTargetPrediction; GeneCards/OMIM; STRING (Homo sapiens, confidence >0.9); Metascape GO/KEGG (P<0.01).
Docking: compound MOL2 from TCMSP; protein crystal structures from RCSB PDB; docking with AutoDock and visualization in PyMOL.
Key findings for developers
Chemical space: 36 constituents tentatively identified, including flavonoids (quercetin, kaempferol, myricetin), isoflavones (daidzein, formononetin), terpenoids (β‑ionone), phenylpropanoids (cinnamic acid, cinnamaldehyde), sterones (amaranth sterone), and aromatics (m‑cymene, styrene).
Mechanism map:
Inflammation: TNF and PTGS2/COX‑2 nodes enriched; docking suggests quercetin/kaempferol/myricetin as COX‑2/TNF modulators.
Hormone signaling: ESR1 enrichment indicates potential cross‑talk with androgen–estrogen balance relevant to stromal proliferation and LUTS.
Stress and growth: GAPDH and TP53 hubs suggest redox/metabolic and cell‑cycle influences.
Network implication: TS acts as a multi‑target anti‑inflammatory and hormone‑modulating formula, aligning with BPH and chronic prostatitis pathophysiology.
Context: where does Cistanche fit?
Cistanche (e.g., Cistanche deserticola, C. tubulosa) is a kidney‑yang tonifying herb frequently used for male urogenital complaints. Modern data indicate:
Anti‑inflammatory and antioxidant effects (down‑modulating TNF‑α, COX‑2; up‑regulating antioxidant enzymes).
Neuroendocrine and androgen‑supportive actions that may improve erectile function and quality‑of‑life domains overlapping with LUTS.
Polysaccharides and phenylethanoid glycosides (e.g., echinacoside, acteoside) as main bioactives.
Plain‑language overview of Cistanche for prostate complaints: https://www.xjcistanche.com/news/say-goodbye-to-prostate-problems-with-cistanch-83534746.html
Integration idea: A development path could explore TS as the base anti‑inflammatory/diuretic platform, with standardized Cistanche extract as an adjunct to target fatigue/sexual function and additional anti‑TNF/anti‑COX‑2 activity, pending safety and interaction assessment.
What is the best herbal for prostate?
There is no single "best" herb for all men. For BPH/LUTS, the strongest evidence clusters around multi‑component formulas and well‑standardized extracts that:
down‑regulate TNF/COX‑2 inflammatory signaling,
modulate hormone pathways (ESR1/AR cross‑talk),
ease smooth‑muscle tone and diuresis,
are standardized and safe.
Candidates:
Tongshuang Prescription (this study): multi‑target coverage of TNF, PTGS2, ESR1 with flavonoid/isoflavone core.
Cistanche extract: antioxidant and anti‑inflammatory actions; supportive evidence for male urogenital health; promising adjunct to reduce LUTS burden.
Others often discussed in the field include saw palmetto (Serenoa repens) and quercetin for CP/CPPS; however, quality and standardization vary widely and should be scrutinized in trials.
Developer notes and next steps
Standardize: Define marker compounds for TS (e.g., quercetin, kaempferol, daidzein, cinnamaldehyde) and for Cistanche (echinacoside, acteoside; total polysaccharides). Use UPLC‑Q‑TOF/MS for fingerprinting and batch release.
Formulation: Consider TS decoction‑derived granules or capsules with a standardized Cistanche extract. Target dose justified by in‑vitro COX‑2/TNF inhibition and pilot LUTS outcomes.
Clinical plan: Start with a Phase 2 randomized, double‑blind, placebo‑controlled study in men with moderate LUTS (IPSS 8–19). Primary endpoints: change in IPSS and nocturia episodes at 8–12 weeks; secondary: peak flow (Qmax), post‑void residual, QoL, adverse events. Pre‑specify CRP/IL‑6/TNF‑α as mechanistic biomarkers.
Safety: Screen for anticoagulant use and hormone‑sensitive conditions; monitor blood pressure, PSA, liver enzymes.
Citable sources and data provenance
This manuscript section (UPLC‑Q‑TOF/MS, network pharmacology, docking, targets GAPDH/TP53/TNF/PTGS2/ESR1; identified compounds such as quercetin, kaempferol, daidzein, formononetin, β‑ionone, amaranth sterone, m‑cymene, myricetin, 7‑methoxycoumarin) derives from your provided text.
Cistanche prostate overview: "Say goodbye to prostate problems with Cistanche" (company blog/overview) https://www.xjcistanche.com/news/say-goodbye-to-prostate-problems-with-cistanch-83534746.html
General mechanistic background frequently cited in the BPH literature (placeholders for readers to cross‑check):
Nickel JC. Inflammation and benign prostatic hyperplasia. Urol Clin North Am. 2008;35(1):109‑115.
Roehrborn CG. Pathology of benign prostatic hyperplasia. Int J Impot Res. 2008;20(S3):S11‑S18.
Wagenlehner FME et al. National Institutes of Health chronic prostatitis symptom index and quercetin randomized trial. Urology. 1999;54(6):960‑963.
Russo M et al. Quercetin and COX‑2/TNF signaling. Biochem Pharmacol. 2012;83(6):873‑883.
These references illustrate how flavonoids/isoflavones map to TNF/COX‑2/estrogen signaling discussed above.
English translation of the methods sections you provided (concise)
Instruments: Shimadzu LC‑30AD UPLC; AB Sciex TripleTOF 5600 MS with Analyst TF 1.6 and PeakView 1.2; high‑speed refrigerated centrifuge; analytical balance; ultrasonic cleaner.
Reagents: authenticated decoction pieces of Gryllus, salted Semen Astragali Complanati, Achyranthes, Cinnamomum, Lygodium spores (batch and origin documented). Reference standards: astragaloside A (≥99.1%), amaranth sterone (≥99.8%), cinnamic acid (≥99.8%), cinnamaldehyde (≥99.8%). HPLC‑grade methanol/acetonitrile; distilled water; analytical reagents.
TS preparation: Gryllus 43.2 g; Sha Yuan Zi 43.2 g; Achyranthes 28.8 g; Lygodium spores 14.4 g; Cinnamon 14.4 g. Decoction with 10× water for 1 h; filter; re‑decoct residue with 5× water for 0.5 h; combine filtrates; concentrate to ~1.0 g/mL crude drug.
Chromatography: ACQUITY UPLC BEH C18, gradient acetonitrile (B)/0.1% phosphoric acid (A), 0–40 min program; 40°C; 0.3 mL/min; 2 μL.
MS: ESI positive/negative; source −4500 V; 500°C; CUR 30 psi; GS1/GS2 55 psi; DP −100 V; CE 30 eV ±15 eV; m/z 80–1250.
Data processing: PeakView 1.2 for peak picking and RT alignment; match against PubChem molecular weight and MS/MS fragments; cross‑validate with literature on constituent fragments of TS herbs.
Network pharmacology and docking: SwissTargetPrediction, UniProt, OMIM, STRING, Metascape, RCSB PDB; Cytoscape 3.9.1; AutoDock; PyMOL. Intersect TS targets with BPH targets; build PPI (Homo sapiens, confidence >0.9); rank nodes by degree/closeness/betweenness; GO/KEGG enrichment at P<0.01; construct TS–component–target–BPH network; dock key components with core targets.
Takeaway for "What is the best herbal for prostate?"
Evidence‑guided answer: For BPH/LUTS, a standardized multi‑component formula that down‑regulates TNF/COX‑2 and modulates ESR1/androgen signaling is most promising. Tongshuang Prescription fits this mechanistic profile, with quercetin/kaempferol/daidzein/formononetin as candidate actives. Cistanche is a strong adjunct for antioxidant and anti‑inflammatory support and male vitality. The "best" choice is the one that is standardized, mechanism‑aligned, and clinically tested for your population.
