Benign prostatic hyperplasia (BPH) is a common benign disease among elderly men [1]. Its pathological features include the proliferation of epithelium, fibromuscular tissue, glands, and stromal cells in the transition zone of the prostate and the perurethral area, which in turn leads to benign enlargement of the prostate. This condition is often accompanied by bladder outlet obstruction and clinically manifests as lower urinary tract symptoms (LUTS) [2]. The combination of these two is collectively referred to as BPH/LUTS. Specifically, LUTS includes frequent urination, urgent urination, urinary incontinence, increased nocturia, delayed urination, poor urine flow, and a feeling of incomplete bladder emptying [3]. These symptoms not only significantly reduce patients’ quality of life but also easily trigger negative emotions, impair physical and mental health, and affect family and social harmony. From an epidemiological perspective, the incidence of BPH shows a significant positive correlation with age. The incidence rate is approximately 50% among men over 50 years old, reaches 83% among those over 80 years old, and about 90% of men over 90 years old have varying degrees of prostatic hyperplasia. the incidence rate among men over 40 years old is around 8% [4]. With the intensification of population aging, clinical attention to this condition has continued to increase. Currently, clinical treatment mainly relies on surgery and medications. The first-line pharmacotherapeutic regimen for BPH/LUTS involves the use of alpha-blockers, 5-alpha-reductase inhibitors, either alone or in combination [5]. However, these medications are not effective for all BPH/LUTS patients, they also tend to cause adverse reactions such as sexual desire disorders and insomnia, and some patients experience drug tolerance issues. These factors limit their clinical application, making it of great significance to explore safe and low-toxicity treatment regimens. In the system of traditional chinese medicine (TCM), BPH is classified under the category of “Long Bi” (dribbling and retention of urine). Its core pathogenesis lies in the dysfunction of bladder qi transformation, with the main disease locations in the kidney and bladder. Syndrome differentiation revolves around kidney deficiency, dampness-heat, and blood stasis [6]. TCM treatment includes internal therapy (oral administration of herbal medicines) and external therapy (such as drug enema, acupuncture, and acupoint application) [7]. It has the advantages of treating both the root cause and symptoms, significant therapeutic effects, and few adverse reactions. TCM treatment not only improves LUTS symptoms in patients but also reduces blood stasis and edema in prostate tissue, thereby assisting in reducing prostate volume [8], which provides an important supplementary approach for BPH treatment.

Vladimiriae Radix is a Tibetan medicine with ethnic characteristics, and it is also a cross-used medicinal species in TCM and Tibetan medicine [9]. Its main producing areas are concentrated in Aba Tibetan autonomous prefecture, Liangshan Yi autonomous prefecture, Xichang, Baoxing, Ya’an (all in Sichuan Province, China), as well as Yunyang, Kaixian, and Nanchuan (all in Chongqing Municipality, China). Its traditional indications align closely with the therapeutic needs of BPH. In Chinese medicine the herb is classified as a qi-regulating drug (pungent, bitter, warm) that promotes flow, assists bladder qi transformation, dispels blood stasis to reduce glandular swelling, and warms to transform damp turbidity in the urethra—directly addressing the “long-bi” (urinary retention) mechanism of BPH. Formulas containing it, such as “Xiang-sha Liu-jun-zi Tang”, have been shown to relieve accompanying symptoms [10]. In Tibetan medicine it is known as “Ma-nu”; it harmonizes the nyes-pa, opens the urethra, disperses the “flesh nodule” of the prostate, and is commonly incorporated into preparations like “Wu-wei Mu-xiang San”. Contemporary clinical studies found that Chinese- and Tibetan-compound prescriptions containing Vladimiriae Radix (e.g., Mu-xiang Dan-shen Yin combined with tamsulosin) can significantly improve IPSS scores, residual urine volume, and maximum urinary flow rate, reduce prostate volume, and cause fewer adverse effects—thereby providing traditional theoretical support for the present study and validating the herb’s candidacy for BPH therapy. The Vladimiriae Radix herb is cylindrical or semi-cylindrical with longitudinal grooves, slightly curved, measuring 10–30 cm in length and 1–3 cm in diameter. Its surface is yellowish-brown or dark brown, with longitudinal wrinkles; where the outer skin peels off, reticulate fine vascular bundles (resembling a loofah sponge) are visible, and the root head occasionally has an “oil head” (a black, sticky gelatinous substance). The herb is light in weight, hard and brittle, and easy to break, the fracture surface is yellowish-white or yellow, containing sparse dark yellow oil spots and cracks (Fig. 1A,B). The xylem is broad with radial textures, and the center of some samples is withered. It has a slight aroma, a bitter taste, and feels sticky when chewed [11]. Modern research has shown that costunolide and dehydrocostus lactone are the core active components in Vladimiriae Radix. Both can significantly inhibit the proliferation of six types of human-derived tumor cells, and the exocyclic double bond at the $\mathrm{\Delta }$11(13) position has been initially found as the main active site of sesquiterpenoid compounds for antitumor activity [12]. Specifically, Bocca et al. [13] showed that costunolide can interact with microtubules to inhibit the proliferation of human breast cancer MCF-7 cells in a dose-dependent manner (with a significant effect at 100 nmol/L) and microtubules may be its new intracellular target. It was reported that costunolide inhibits the proliferation and induces the apoptosis of MCF-7 cells by regulating the expression of apoptosis-related proteins such as Bax, Bcl-2, p53 and Caspase-3 [14]. On the other hand, Roy and Manikkam [15] found that dehydrocostus lactone can inhibit the activity of thioredoxin reductase 1 (TrxR1) in Henrietta Lacks (HeLa) cells with a half-maximal inhibitory concentration (IC₅₀) of 12.00 µmol/L, triggering the accumulation of reactive oxygen species (ROS) and the collapse of redox homeostasis, which ultimately induces cell apoptosis. Therefore, in-depth exploration of the material basis and mechanism of action of Vladimiriae Radix in anti-BPH can provide theoretical support for its rational clinical application and the research and development of related drugs, and thus holds important research significance.

Fig. 1.

Original plant, dried medicinal material and main active components of Vladimiriae Radix. (A) The original plant of Vladimiriae Radix (the plant with rhizomes, roots and leaves). (B) Dried medicinal material of Vladimiriae Radix, Core active components which mainly including (C) costunolide, (D) dehydrocostus lactone, (E) oleanolic acid, (F) quercetin, (G) luteolin, and (H) taraxasterol from Vladimiriae Radix.

To systematically elaborate the scientific mechanism underlying the anti-BPH effect of the dried roots of Vladimiriae Radix, this study employed network pharmacology and molecular docking techniques as core methods. From the three-dimensional perspective of “component-target-pathway”, it explored the pharmacodynamic material basis and action logic of D. souliei. This not only addresses the practical challenges in clinical treatment of BPH but also provides a new paradigm for the modernization research on the integrated application of Tibetan medicine and TCM [17]. BPH, a common benign disease of the urinary system in elderly men, exhibits distinct “multidimensional intertwined” characteristics in its pathological mechanism [18]. Specifically, the uncontrolled proliferation of epithelial and stromal cells in the prostatic transition zone represents the core pathological manifestation [19], the imbalance of the hormonal microenvironment mediated by DHT serves as the key driving factor [20], and chronic non-bacterial inflammatory infiltration and dysregulation of urethral cholinergic nerve modulation respectively exacerbate tissue damage and LUTS [21]. These three factors together form the complex pathological network of BPH. In current first-line clinical treatment regimens, although $\alpha$-adrenergic blockers can rapidly relax urethral smooth muscle to relieve dysuria, they are prone to causing adverse reactions such as orthostatic hypotension and dizziness [22]. While 5$\alpha$-reductase inhibitors can reduce prostate volume by decreasing DHT production, they are associated with issues like decreased libido and drug tolerance [23]. The limitations of such “single-target intervention” [24] highlight the urgency of exploring multi-target, low-toxicity treatment strategies. In TCM, BPH is categorized under the scope of “Long Bi” (dysuria and anuria syndrome). Its core pathogenesis is defined as “kidney deficiency as the root cause, dampness-heat as the superficial symptom, and blood stasis as the pathological change” [25]. Notably, the characteristic of TCM involving the synergistic effect of multiple components is highly compatible with the pathological complexity of BPH. Vladimiriae Radix, a characteristic medicinal herb used in Tibetan medicine for “promoting qi circulation and dredging collaterals” and in TCM for “strengthening the spleen and harmonizing the stomach”, has been the focus of previous studies mostly on the analgesic, anti-inflammatory, and anti-tumor activities of its sesquiterpene lactone components. However, a systematic understanding of its mechanism of action against BPH has not yet been established, which also serves as the core research direction of this study [26]. In the present study, through the combined retrieval of multiple databases including TCMSP, PubChem, and HERB, and combined with the strict evaluation of gastrointestinal absorption rate, DL, Analytical data ensuring that the six finalized compounds (costunolide, dehydrocostus lactone, luteolin, quercetin, oleanolic acid, taraxasterol) possess favorable pharmacokinetic profiles. This provides a clear direction for the material basis of Vladimiriae Radix in anti-BPH. Among these components, costunolide and dehydrocostunolide, as the sesquiterpene lactone components with the highest content in Vladimiriae Radix [27], have the $\mathrm{\Delta }$11(13) exocyclic double bond in their molecular structures, which has been found to be a key functional domain for regulating cell proliferation. Previous studies have shown that these two components can inhibit the proliferative activity of tumor cells such as breast cancer MCF-7 and cervical cancer HeLa by disrupting the stability of microtubule polymerization and inducing the ROS-mediated apoptotic pathway. Considering the commonality in the pathological feature of “uncontrolled cell proliferation” between BPH and tumors [28], it is hypothesized that these components may target and inhibit the abnormal proliferation of prostate cells through similar molecular mechanisms. This hypothesis has also been initially supported by the subsequent molecular docking results. In addition, oleanolic acid, a pentacyclic triterpenoid component, can reduce the release of inflammatory factors such as IL-6 and tumor necrosis factor-alpha (TNF-$\alpha$) by inhibiting the nuclear factor-kappa B (NF-$\kappa$B) inflammatory signaling pathway, thereby specifically improving the chronic inflammatory microenvironment of BPH [29]. Quercetin, a flavonoid component, can correct the imbalance of estrogen and androgen ratios in prostate tissue by regulating the activity of estrogen receptors [30]. Meanwhile, phytosterols such as stigmasterol and $\beta$-sitosterol can indirectly regulate the local hormone levels in the prostate by interfering with cholesterol metabolism (the precursor pathway for steroid hormone synthesis) [31]. These 6 components cover the three core pathological links of BPH, namely “proliferation inhibition, inflammation alleviation, and hormone regulation”. This not only reflects the diversity of the pharmacodynamic substances of Vladimiriae Radix but also echoes the concept of the “monarch-minister-adjuvant-courier” synergistic effect in TCM. The excavation and validation of core targets further suggest the pivotal role of Vladimiriae Radix against BPH. Through database cross-analysis and the construction of a PPI network, this study screened 235 overlapping targets from 342 active targets of Vladimiriae Radix and 5256 disease targets of BPH. Based on degree value and topological analysis, 6 core targets (AR, CYP17A1, CYP19A1, ACHE, F2, and HMGCR) were identified, and these targets exactly correspond to the key nodes in the pathological mechanism of BPH. Among these, the AR is the “hub target” for BPH pathogenesis [32]. Specifically, the specific binding of DHT to AR can activate the expression of downstream proliferation-related genes such as MYC and CCND1, thereby promoting the continuous proliferation of prostate cells. Clinically, the drug finasteride achieves “indirect regulation” of AR by inhibiting 5$\alpha$-reductase to reduce DHT production [33]. However, the molecular docking results of this study showed that costunolide and dehydrocostunolide can form hydrogen bonds, hydrophobic interactions, and covalent bonds with active sites such as LEU-93, ILE-119, and TRP-189 of AR. This suggests that these components may directly block the formation and nuclear translocation of the DHT-AR complex by competitively binding to the ligand-binding domain of AR. Compared with clinical drugs, this “direct intervention” mode may have stronger target specificity. Cytochrome P450 enzyme family members CYP17A1 and CYP19A1 jointly regulate steroid hormone metabolism [34], CYP17A1, as the rate-limiting enzyme in androgen synthesis, an abnormal increase in its activity leads to local androgen accumulation in the prostate [35], CYP19A1 (aromatase) can convert testosterone to estradiol, and the imbalance of estrogen and androgen ratios induced by its increased activity further promotes AR expression and stromal cell proliferation [36]. This study found that Vladimiriae Radix components can bind to the active sites of CYP17A1 (such as PHE-446 and CYS-453) and CYP19A1 (such as ALA-306 and ARG-375). It is hypothesized that these components may correct hormone imbalance at the source by exerting dual inhibition on the activity of these two enzymes. Compared with abiraterone, which only inhibits CYP17A1, this “dual-target regulation” mode may reduce the risk of adverse reactions while regulating hormone balance. The identification of ACHE, F2, and HMGCR further expands the mechanistic dimensions of Vladimiriae Radix in anti-BPH. ACHE maintains cholinergic nerve signal balance by degrading acetylcholine in the synaptic cleft [37]. Abnormal elevation of its activity leads to excessive contraction of urethral smooth muscle, which exacerbates bladder outlet obstruction and LUTS. This study shows that Vladimiriae Radix components can bind to the catalytic active sites of ACHE (such as CYS-345 and SER-347). It is hypothesized that these components may increase acetylcholine concentration by inhibiting ACHE activity, thereby enhancing the regulation of urethral smooth muscle relaxation. This mechanism is complementary to that of solifenacin, a clinical anticholinergic drug, and the nature of natural components may reduce the risk of central nervous system adverse reactions. Factor Ⅱ (F2), a key factor in the coagulation-inflammation pathway, can promote the release of inflammatory factors by activating protease-activated receptors (PARS), thereby exacerbating inflammation and fibrosis in prostate tissue [38]. As the rate-limiting enzyme in cholesterol synthesis, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) not only affects steroid hormone synthesis, but its abnormal activity is also closely associated with oxidative stress and cell proliferation in prostate tissue [39]—clinical studies have found that statins can reduce the risk of BPH development by inhibiting HMGCR. The good binding activity of Vladimiriae Radix components to F2 and HMGCR suggests that these components may intervene in the progression of BPH from two dimensions—the inflammatory microenvironment and metabolic abnormalities—by regulating the coagulation-inflammation pathway and lipid metabolism pathway. This further improves the “multi-target synergy” mechanism of Vladimiriae Radix. GO functional annotation and KEGG pathway enrichment analyses preliminarily corroborated, at the systems level, the multi-dimensional characteristics of Vladimiriae Radix against BPH. At the BP level, core targets are enriched in terms such as steroid hormone response and vascular processes, which are highly consistent with the hormone-dependent characteristics of BPH and the mechanism of tissue angiogenesis. At the CC level, targets are concentrated in structures including membrane rafts and membrane microdomains. As aggregation platforms for signaling molecules (e.g., AR, G protein-coupled receptors), dysfunction of membrane rafts is a key link in the loss of control of BPH pathological signals—suggesting that Vladimiriae Radix components may inhibit signal transmission by interfering with membrane raft structure [40]. At the MF level, G-protein-coupled amine receptor activity and acetylcholine receptor activity were significantly enriched, further suggesting that Vladimiriae Radix intervenes in BPH by modulating receptor activity. The results of KEGG pathway enrichment analysis showed that the core targets are mainly involved in pathways such as lipid and atherosclerosis, cholinergic synapses, and AGE-RAGE. Specifically, lipid metabolism disorders can exacerbate BPH through cholesterol deposition, the cholinergic synapses pathway directly corresponds to the mechanism of action of ACHE [41], and the AGE-RAGE pathway is closely associated with BPH tissue fibrosis [42]. The synergistic enrichment of these pathways fully reflects the characteristic of Vladimiriae Radix in intervening in BPH through a “multi-pathway and multi-link” manner, and also provides a modern biological explanation for the advantage of “treating both the root cause and symptoms” (a key principle of TCM). Molecular docking results further support the inference that the active components of Vladimiriae Radix possess specific binding capabilities to the core targets. The binding sites of components such as costunolide and dehydrocostunolide with targets including AR, CYP17A1, and ACHE are all located in the active center regions of these targets, and binding is achieved through stable interactions such as hydrogen bonds, hydrophobic interactions, and salt bridges. For instance, there is a hydrogen bond formed between costunolide and TRP-189 of AR, and a salt bridge formed between costunolide and ARG-109 of CYP17A1. Both the type and strength of these interactions meet the key indicators of molecular binding activity, providing a basis for Vladimiriae Radix components to serve as candidate compounds for BPH treatment. Differences in the binding modes between different components and the same target (e.g., the number of hydrogen bonds formed between quercetin and ACHE is greater than that between costunolide and ACHE) suggest that there may be potential for synergistically enhancing target regulation among these components. This is also consistent with the characteristic of the synergistic effect of multiple components in TCM. The innovative value of this study is mainly reflected in two aspects: Firstly, it constructs the “component-target-pathway” systematic network of Vladimiriae Radix for anti-BPH from the perspective of network pharmacology for the first time, clarifies its core active components and key functional nodes, and fills the research gap of Vladimiriae Radix in the field of BPH treatment [43]. Secondly, it realizes the cross-integration of Tibetan medicine characteristic medicinal materials and modern pharmacology technologies, and provides reference methodological ideas for the mechanism research of ethnic medicine in the treatment of urinary system diseases. However, this study still has limitations. Firstly, network pharmacology relies on the integrity of existing databases, which may miss components with low content but high activity or unannotated targets. Secondly, molecular docking is only an in vitro computer simulation, lacking functional verification from in vivo BPH animal models and in vitro cell experiments, and thus cannot clarify the actual regulatory effect of core components on targets. Furthermore, the impact of Vladimiriae Radix processing techniques on active components has not been incorporated into the study. Processing is a crucial link for TCM to exert its efficacy and may alter its pharmacodynamic material basis. In summary, the present study preliminarily proposes that Vladimiriae Radix exerts anti-BPH effects through the synergistic action of “multi-component, multi-target, and multi-pathway” mechanisms, identifying its core active ingredients and key therapeutic targets. These findings provide experimental support for both basic research and clinical application of Vladimiriae Radix, and offer a new perspective for investigating the mechanisms of traditional Chinese medicine in BPH treatment. Further experimental validation is needed to refine this mechanism and promote the translational application of Vladimiriae Radix in BPH therapy.

Costunolide, dehydrocostus lactone, oleanolic acid, luteolin, quercetin, and taraxasterol are bioactive components of Vladimiriae Radix with potential anti-BPH properties. AR, CYP17A1, CYP19A1, ACHE, F2, and HMGCR were the core targets which related to the bioactive components in anti-BPH. Oleanolic acid exhibits the good binding energy with 6 core targets. Cell-level verification showed that oleanolic acid may significantly inhibits the proliferation of DHT-induced BPH-1 cells (p 0.05) and regulates the mRNA expression of core targets (downregulating AR and HMGCR, upregulating CYP17A1, CYP19A1, and ACHE, with no significant effect on F2). In summary, the study found that Vladimiriae Radix exerts anti-BPH effects through a “multi-component, multi-target, and multi-pathway” mode of action, filling the gap in the anti-BPH mechanism research of Vladimiriae Radix and providing reliable experimental evidence for the deepening of its basic research, promotion of clinical application, and reference for the modernization of Tibetan-Chinese integrated medicinal materials.

The datasets generated and analysed during the current study are available upon request from the corresponding author.

JXM conceived and designed the research. XDF, JW, and JXM carried out the data analysis and wrote the paper in the present study. XDF and JW finished the drawing and experiment. JXM carried out the manuscript revision work. All authors participated in the editing and revision of the manuscript. All authors have read and approved the final manuscript. All authors fully participated in this research and agree to be accountable for all aspects of the study.

Not applicable.

Not applicable.

This work was financially supported by Key Scientific and Technological Research Project of Chongqing Municipal Education Commission (No. KJZD-K202302801), Scientific Research Project of Chongqing Medical and Pharmaceutical College (ygz2022104, ygzrc2024104, ygzrc2024101), Chongqing Municipal Education Commission Youth Project (KJQN202402816) respectively.

The authors declare no conflict of interest.