Metabolomics Combined With BMP-2/Smad Signaling Pathway Analysis Elucidates The Ameliorative Effects Of Total Flavonoids Of Epimedii Folium—Bone Nutritional Supplement On Postmenopausal Osteoporosis
Apr 29, 2026
Abstract
Postmenopausal osteoporosis (PMOP) is triggered by estrogen deficiency. Insufficient estrogen leads to bone resorption exceeding bone formation, thereby causing decreased bone mass. Studies have confirmed that the total flavonoids extract from Epimedii Folium (TFE) can ameliorate osteoporosis, while a bone nutrient consisting of chondroitin sulfate (CS) combined with glucosamine hydrochloride (GLcN) exerts therapeutic effects on bone metabolism–related diseases. Since osteoporosis is fundamentally characterized by metabolic disorders, the combination of TFE extract with a bone nutritional supplement (TG) may exert synergistic multi-target regulatory effects, thereby offering a new approach for the prevention and control of PMOP.
This study systematically explored the ameliorating effects and mechanisms of TG on PMOP through in vivo and in vitro experiments combined with metabolomics technology. In vivo, an ovariectomized (OVX) rat model was established by bilateral ovariectomy. The pharmacodynamic effects of TG on PMOP were verified based on bone histopathological staining, bone mineral density, and physiological indicators. In addition, metabolomics technology was used to quantify urine metabolites to explore potential PMOP biomarkers regulated by TG and the associated metabolic networks. In vitro, MC3T3-E1 cells were treated with TFE and TG to screen safe doses. Subsequently, osteogenic induction was performed, and the effects and mechanisms of TG on osteogenic differentiation were studied via alkaline phosphatase (ALP) staining, Alizarin Red S (ARS) staining, and Western blot analysis.
Pharmacodynamic results showed that TG reduced osteoclast generation and significantly restored bone mineral density and physiological indicator levels. Metabolomics results indicated that TG alleviated bone loss mainly by intervening in multiple metabolic pathways, including nicotinate and nicotinamide metabolism, histidine metabolism, pyrimidine metabolism, tryptophan metabolism, alanine/aspartate/glutamate metabolism, purine metabolism, vitamin B6 metabolism, and others. In vitro experiments confirmed that TG promoted osteoblast proliferation and osteogenic differentiation via the bone morphogenetic protein-2 (BMP-2)/Sma- and Mad-related protein (Smad) signaling pathway. In summary, TG can effectively ameliorate PMOP through a multi-target synergistic effect. This study provides an experimental basis and theoretical support for the clinical application of TG.
Key words: total flavonoids of Epimedii Folium; bone nutritional supplement; postmenopausal osteoporosis; metabolomics; BMP-2/Smad signaling pathway
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1. Introduction
Osteoporosis (OP) is a systemic metabolic bone disease characterized by decreased bone mass and bone mineral density (BMD), destruction of bone microarchitecture, and increased bone fragility. Postmenopausal osteoporosis (PMOP) is the most common type and is associated with reduced estrogen secretion. Decreased estrogen causes bone resorption to exceed bone formation, resulting in disrupted bone homeostasis [1].
A China osteoporosis prevalence study including 20,416 participants found that among people aged 40 years or older, the prevalence of OP was 5.0% in men and 20.6% in women [2]. An evaluation of OP prevalence among U.S. adults aged ≥50 years found prevalence rates of 16% in men and 29.9% in women [3], indicating a higher prevalence in women. Clinically, drugs used to treat PMOP mainly include parathyroid hormone analogs, bisphosphonates, estrogen, calcitonin, and RANKL inhibitors; however, these drugs may cause adverse effects such as palpitations, osteonecrosis of the jaw, and cardiovascular disease, making them unsuitable for long-term use [4]. Due to the notable side-effect burden of conventional drugs, identifying traditional Chinese medicine (TCM) options with fewer adverse reactions and suitability for long-term management has become urgent. TCM has long been used to improve OP and has received broad attention domestically and internationally. Studies confirm that multiple Chinese herbs such as Epimedii Folium and Rhizoma Drynariae have significant effects in improving OP [5].
Total flavonoids of Epimedii Folium (TFE) are the main active components, including icariin, epimedin A, and epimedin B. The anti-osteoporotic effects of Epimedii Folium have been reported in many studies; for example, icariin enhances osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) [6]. TFE extract can also improve bone loss by increasing brown adipose tissue and reducing white adipose tissue in bone marrow [7].
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Chondroitin sulfate (CS) is a sulfated glycosaminoglycan and a linear polysaccharide composed of repeating disaccharide units containing uronic acid and N-acetylhexosamine [8]. Glucosamine hydrochloride (GLcN) is an amino monosaccharide synthesized from glucose in human tissues; it is mainly present in cartilage and synovial connective tissue and is an important component of articular cartilage and extracellular matrix. Both are commonly used as dietary supplements. ZHENG H X et al. [9] reported that CS can reduce serum inflammatory cytokines and inhibit oxidative stress; it may also reduce osteoclast expression by regulating OPG/RANKL, suggesting anti-inflammatory and antioxidant actions and inhibition of osteoclast differentiation. In the United States, CS is often combined with GLcN in dietary supplements and is mainly used to reduce the loss of matrix components in osteoarthritis [10]. Moreover, because CS can induce cell differentiation, it has been used in chondrogenesis and osteogenic constructs [11–12].
Multiple studies suggest that TFE extract, CS, and GLcN have certain therapeutic effects in bone metabolism–related diseases. However, whether a combined TFE extract–bone nutrient supplement (TG) produces synergistic benefits in improving OP and its mechanisms remain unclear.
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2. Materials and Methods
2.1 Animals
Forty 8-week-old female Sprague–Dawley rats (180–220 g) were purchased from Changchun Yisi Experimental Animal Co., Ltd. (Animal license No. SCXK (Ji)-2020-0001). Animals were housed in an SPF laboratory at 23±223 \pm 223±2°C and 50% humidity, with free access to food and water, under a 12 h light/12 h dark cycle. Animal experiments were reviewed and approved by the Experimental Animal Welfare and Ethics Committee of Jilin Agricultural University (Acceptance No. 20240717001).
2.2 Drugs and Reagents
Korean Epimedii Folium crude herb was purchased from Tonghua County Senyu Chinese Medicinal Materials Co., Ltd. (Jilin Province). Icariin (Batch No. T11A11B111118) was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. GLcN (Batch No. M-AT-1904020) was purchased from Zhejiang Jinke Pharmaceutical Co., Ltd. CS (Batch No. HS1904270) was purchased from Jiaxing Hengjie Biopharmaceutical Co., Ltd. Additional reagents included penicillin sodium for injection, estradiol valerate tablets, tribromoethanol, ELISA kits for PINP and CTX-I, hydroxyproline, phosphorus and calcium assay kits, ALP staining kit, α-MEM medium, trypsin, penicillin-streptomycin, blocking buffer, TBST, rapid transfer buffer, electrophoresis buffer, PVDF membranes, HRP goat anti-rabbit IgG, anti-GAPDH antibody, pre-stained protein marker, RIPA lysis buffer, ECL kit, PAGE gel preparation kit, protein loading buffer, CCK-8 kit, fetal bovine serum, Alizarin Red S, ascorbic acid, β-glycerophosphate sodium, dexamethasone, PBS, and primary antibodies for RUNX2, BMP-2, Smad1, ALP, OSX, and p-Smad1/5. (All batch numbers and suppliers are as listed in the original text.)
2.3 Instruments
A microplate reader (ThermoFisher, USA), inverted microscope (Motic Group), biosafety cabinet (ThermoFisher, USA), CO2_22 incubator (ThermoFisher, USA), and digital dual-channel heating metal bath (Changchun Kelebo Biotechnology Co., Ltd.) were used.
2.4 TFE Extraction
TFE was extracted using the method reported by Zhang Dongxue et al. [13]. The obtained lyophilized powder was measured by UV–Vis spectrophotometry at 270 nm. Using icariin as the standard, the total flavonoid content of the TFE extract was determined to be 79.0%.
2.5 Animal Experimental Design
Rats were randomized into five groups: normal (sham), model (OVX), estradiol valerate (EV), TFE extract (T), and TG. The sham group underwent removal of a small amount of abdominal fat and soft tissue only; all other groups underwent bilateral ovariectomy. Drugs were prepared as suspensions in 0.5% sodium carboxymethyl cellulose and administered by oral gavage. Sham and OVX groups received saline by gavage. The T group received TFE at 490 mg·kg−1^{-1}−1. The EV group received 0.1 mg·kg−1^{-1}−1. The TG group received CS 124 mg·kg−1^{-1}−1, GLcN 155 mg·kg−1^{-1}−1, and TFE 490 mg·kg−1^{-1}−1. Doses were converted from human recommended doses. Gavage volume was 2 mL·kg−1^{-1}−1·d−1^{-1}−1 for 8 consecutive weeks. Successful modeling was confirmed when serum estradiol (E2) levels in the OVX group were significantly lower than those in the sham group. Because CS+GLcN alone showed only a slight improving trend in rat BMD, a separate bone nutrient–only group was not established.
At weeks 3, 6, and 8, rats were placed in metabolic cages with free water and 12 h fasting, and 24 h urine samples were collected. Urine was centrifuged at 4°C, 3,500 r·min−1^{-1}−1 for 10 min; supernatants were stored at −80°C. At the end of week 8, animals were euthanized and blood was collected, centrifuged at 4°C, 4,000 r·min−1^{-1}−1 for 15 min to obtain serum, then stored at −80°C.
2.6 Measurement of Physiological Indicators
At week 8, urine and abdominal aortic blood were collected. ELISA kits were used to measure serum bone formation marker PINP and bone resorption marker CTX-I. Serum and urine hydroxyproline (Hyp) were measured by alkaline hydrolysis. Urine calcium (U-Ca) was measured by the MTB microplate method, and urine phosphorus (U-P) by the phosphomolybdate method.
2.7 Bone Mineral Density and Micro-CT
After 8 weeks, the left femurs were harvested, cleaned, and scanned using micro-CT. Trabecular parameters were quantitatively analyzed, including trabecular separation (Tb.Sp), trabecular thickness (Tb.Th), trabecular number (Tb.N), bone volume fraction (BV/TV), and structure model index (SMI).
2.8 Bone Histopathology
Right femurs were fixed in 4% paraformaldehyde, decalcified, paraffin-embedded, sectioned, and stained with HE and TRAP. TRAP-positive osteoclasts appeared dark red.
2.9 Metabolomics Analysis
Urine (100 μL) was diluted 10-fold with distilled water, vortexed 30 s, centrifuged at 4°C, 15,000 r·min−1^{-1}−1 for 10 min, and the supernatant was used for analysis. Equal volumes (100 μL) of urine from each rat were pooled to prepare quality control (QC) samples.
Chromatography was performed using an ACQUITY UPLC CSH C18 column (2.1 mm × 150 mm, 1.7 μm) at 40°C. Mobile phase A was acetonitrile (0.1% formic acid) and mobile phase B was water (0.1% formic acid). Flow rate was 0.20 mL·min−1^{-1}−1, injection volume 5 μL. Gradient: 0–4 min, 95% B; 4–5 min, 95%–65% B; 5–13 min, 65%–25% B; 13–14 min, 25%–0% B; 14–18 min, 0%–95% B.
Mass spectrometry was performed using a Q Exactive Plus instrument in positive and negative ion modes with spray voltage ±3.2 kV, sheath gas 2.0–8.0 psi, capillary temperature 350°C, and scan range m/z 100–1,000.
2.10 Multivariate Statistics and Data Processing
Raw LC–MS data were preprocessed and normalized using Compound Discoverer software. Unsupervised PCA was used to assess clustering and group differences. Supervised OPLS-DA was used to obtain VIP values. A t-test was applied to normalized data. Metabolites with VIP > 1 and P < 0.05 were considered potential biomarkers and further identified.
2.11 Biomarker Identification and Pathway Analysis
Accurate retention time (Rt), m/z, and molecular weight were obtained from TIC chromatograms. Candidate formulas and elemental composition were predicted using Compound Discoverer online databases (HMDB, KEGG). Pathway enrichment was performed using MetaboAnalyst.
2.12 Osteoblast Culture and Differentiation
MC3T3-E1 mouse calvarial osteoblasts were cultured in a 37°C CO2_22 incubator. When 80%–90% confluent, cells were plated for osteogenic induction. Osteogenic complete medium consisted of α-MEM supplemented with 10 mmol·L−1^{-1}−1 β-glycerophosphate, 50 μmol·L−1^{-1}−1 ascorbic acid, and 100 nmol·L−1^{-1}−1 dexamethasone.
2.13 Cytotoxicity Assay
Cell viability was assessed by CCK-8. MC3T3-E1 cells (5 × 103^33 cells/well) were seeded in 96-well plates and incubated overnight. Cells were then treated with TFE (2–12 μg·mL−1^{-1}−1), CS (50–250 μg·mL−1^{-1}−1), or GLcN (0.2–1.0 mmol·L−1^{-1}−1) for 24 h. Absorbance at 450 nm was measured to calculate viability. Based on results, optimal doses were selected and combined to test TG versus control.
2.14 ALP Staining
MC3T3-E1 cells (1 × 104^44 cells/well) were seeded in 12-well plates and divided into blank (no induction), control (osteogenic induction), T (osteogenic induction + TFE), and TG (osteogenic induction + TG). Media were changed every other day. After 7 days, ALP staining was performed per kit instructions.
2.15 ARS Staining
Using the same grouping and treatment as ALP staining, cells were induced for 14 days. Cells were fixed with 4% paraformaldehyde, stained with 0.1% Alizarin Red (pH 7.2) for 30 min, washed, and observed under a microscope.
2.16 Western Blot
To examine whether TG influences osteogenic differentiation through BMP-2/Smad signaling, MC3T3-E1 cells (5 × 105^55 cells/well) were seeded in 6-well plates with the same grouping as above. Cells were lysed with RIPA plus protease and phosphatase inhibitors. After BCA quantification, proteins were denatured, separated by 10% SDS-PAGE, transferred to PVDF, blocked, incubated with primary antibodies (GAPDH, p-Smad1/5, Smad1, BMP-2, RUNX2, OSX, ALP), followed by HRP secondary antibody, ECL development, imaging, and ImageJ quantification.
2.17 Statistical Analysis
Data are expressed as mean ± SD. SPSS was used for statistical analysis. After normality testing, one-way ANOVA was performed. Differences were considered statistically significant at P < 0.05.
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3. Results
3.1 Serum Biochemical Indicators
As shown in Table 1, compared with the sham group, E2 levels in the OVX group significantly decreased, indicating successful model establishment. Serum PINP in the OVX group significantly decreased, indicating reduced bone formation in PMOP. Compared with OVX, PINP in the T and TG groups significantly increased, indicating that both T and TG increased bone formation markers, with better recovery in TG. Compared with sham, CTX-I in OVX significantly increased, indicating enhanced bone resorption after ovariectomy. Compared with OVX, CTX-I in T and TG significantly decreased, with a more pronounced reduction in TG. Serum hydroxyproline (S-Hyp) significantly increased in OVX versus sham, indicating increased collagen degradation; compared with OVX, S-Hyp decreased in T and TG, with a significant reduction in TG, suggesting better inhibition of collagen degradation.
3.2 Urine Biochemical Indicators
Compared with sham, U-P significantly increased in OVX; compared with OVX, U-P decreased more markedly in TG. U-Ca significantly increased in OVX versus sham; compared with OVX, U-Ca significantly decreased in both T and TG, with a larger decrease in TG. Urine Hyp (U-Hyp) significantly increased in OVX versus sham; compared with OVX, U-Hyp significantly decreased in each treatment group, with a greater decrease in TG than in T (Table 2).
3.3 Bone Histopathology
HE staining showed that trabecular continuity was good in sham, with fewer adipocytes in the marrow cavity. Compared with sham, OVX showed poorer trabecular continuity and markedly increased adipocytes. Compared with OVX, trabecular continuity recovered in T and TG and adipocyte numbers decreased; TG showed more obvious recovery (Figure 1A). TRAP staining showed that osteoclast numbers were significantly increased in OVX versus sham; compared with OVX, osteoclasts decreased in T and TG, with a significant decrease in TG (Figure 1B).
3.4 Bone Mineral Density and Micro-CT Parameters
Micro-CT showed significantly reduced bone mass in OVX compared with sham. Compared with OVX, bone mass recovery in TG was more significant than in T, with a clear increase in trabecular density (Figure 1C). Quantitative analysis showed BV/TV, Tb.Th, and Tb.N significantly decreased in OVX versus sham. Compared with OVX, these indices significantly increased after TG (better than T). Tb.Sp and SMI significantly increased in OVX versus sham; compared with OVX, TG decreased these indices more significantly than T (Table 3).
3.5 PCA
Under both positive and negative ion modes, PCA of urine metabolite profiles at weeks 3, 6, and 8 showed clear separation among sham, OVX, and TG groups, indicating distinct metabolic profiles before/after modeling and before/after TG treatment (see supplementary material referenced in the original publication context).
3.6 Potential Biomarker Screening
OPLS-DA showed more pronounced group separation across weeks in both ion modes. Volcano plots and 200-permutation tests supported model reliability (R2Y and Q2 close to 1; Q2 intercept < 0), suggesting no overfitting.
3.7 Biomarker Identification and Pathway Analysis
Using VIP > 1 and P < 0.05, potential biomarkers were screened. Based on m/z, MS/MS, Rt, and molecular weight, and referencing HMDB/KEGG, 46 potential biomarkers were identified. KEGG enrichment indicated pathways mainly involved nicotinate and nicotinamide metabolism, histidine metabolism, pyrimidine metabolism, tryptophan metabolism, D-amino acid metabolism, alanine/aspartate/glutamate metabolism, purine metabolism, and vitamin B6 metabolism (Figure 2). Hierarchical clustering showed dynamic changes across weeks 3, 6, and 8 after TG intervention (see supplementary material referenced in the original publication context).
3.8 Cytotoxicity Assay
After 24 h treatment, 6 μg·mL−1^{-1}−1 TFE increased cell viability; higher doses inhibited viability. GLcN at 0.4 mmol·L−1^{-1}−1 significantly increased viability. CS at 150 μg·mL−1^{-1}−1 increased viability (Table 4). Thus, 6 μg·mL−1^{-1}−1 TFE, 0.4 mmol·L−1^{-1}−1 GLcN, and 150 μg·mL−1^{-1}−1 CS were selected. Combined dosing showed control viability of 1.00 ± 0.11 and TG viability of 1.22 ± 0.03, indicating the selected TG dose was safe and non-cytotoxic.
3.9 Effects on Osteogenic Differentiation and Protein Expression
ALP is an early marker of osteogenic differentiation. Under osteogenic induction, TG showed darker ALP staining than T, indicating better promotion of ALP expression (Figure 3A). ARS staining showed that both T and TG promoted mineralized nodule formation compared with control, with a more pronounced effect in TG (Figure 3B). Western blot showed increased expression of osteogenic markers under induction. Compared with blank, control indicated successful osteogenic differentiation. After treatment, TG more strongly increased BMP-2, p-Smad1/5, RUNX2, OSX, and ALP protein expression (Figure 3C; Table 5).
4. Discussion
Postmenopausal ovarian failure reduces estrogen levels and disrupts bone metabolism, enhancing osteoclast differentiation while decreasing osteoblast differentiation, ultimately causing bone loss [14–15]. Based on prior research [13], it was hypothesized that combining TFE with a bone nutritional supplement may exert synergistic effects in improving PMOP. Therefore, this study used an OVX rat model and an osteoblast differentiation model to evaluate TG efficacy and mechanism.
PINP (bone formation) and CTX-I (bone resorption) were improved by both T and TG, with TG showing more significant effects. Hydroxyproline (Hyp), a major collagen component, increases in serum and urine in OP [16–17]. In this study, OVX increased both serum and urine Hyp; treatment reduced these levels, with TG reducing them more significantly than T. Increased bone resorption in OVX also increases calcium and phosphorus loss, raising urinary calcium and phosphorus, consistent with DESAI S et al. [18]. Treatment decreased U-Ca and U-P, with greater reductions in TG. Estrogen deficiency alters trabecular structure, inhibits BMSC osteogenesis, promotes adipogenesis, and increases osteoclast activity [19]. TG improved trabecular connectivity, reduced marrow adipocytes, and reduced osteoclast numbers more effectively than T. Micro-CT confirmed that TG increased BMD and improved trabecular microarchitecture (BV/TV, Tb.Th, Tb.N) while reducing Tb.Sp and SMI.
Because OP is a metabolic disease, metabolomics was used to explore metabolic networks regulated by TG. TG's regulated pathways mainly involved amino acid, nucleotide, and vitamin metabolism. Tryptophan is associated with BMD and reduced levels may mark OP [20]. In this study, tryptophan was downregulated in OVX and significantly restored after 8 weeks of TG treatment, suggesting TG may alleviate bone loss by modulating tryptophan metabolism. Creatinine, linked to arginine/creatine metabolism, has been associated with OP [21–22]. Here, creatinine was highest in OVX at week 8 and TG significantly reduced it, suggesting regulation of arginine metabolism may contribute to PMOP improvement. Glutathione metabolism participates in PMOP [23]. L-glutamate can be generated from glutathione (GSH) metabolism. GSH has antioxidant functions and supports bone remodeling [24]. In PMOP, oxidative stress increases, antioxidant capacity decreases, and osteoclast activity exceeds osteoblast activity, impairing bone formation [15]. In this study, L-glutamate was higher in OVX at week 8, and TG decreased L-glutamate, suggesting TG may reduce conversion of GSH to L-glutamate and thereby support GSH levels and counteract estrogen-deficiency–related metabolic abnormalities.
Purine metabolism disorders in OP can lead to uric acid accumulation [25–27]. Metabolomics showed TG reduced uric acid levels at weeks 3, 6, and 8 compared with OVX, suggesting suppression of purine synthesis and uric acid accumulation. LIU D F et al. [28] reported that interventions can increase adenosine to relieve purine metabolism disorders. In this study, TG significantly increased adenosine after 8 weeks, suggesting a contribution to osteogenic differentiation and bone recovery.
Vitamins (D, B complex, K) play important roles in bone metabolism [29–31]. Niacin and nicotinamide are vitamin B3 forms [32]. Niacin contributes to NAD/NADP coenzyme synthesis, with anti-inflammatory and antioxidant effects [33]. Nicotinamide can also promote NAD production; NAD supports mitochondrial function, reduces ROS damage, and helps maintain bone homeostasis. In OP, reduced NAD may hinder osteogenic differentiation and mineralization [34]. In this study, niacin and nicotinamide were higher in OVX at weeks 3, 6, and 8 (highest at week 8). The authors inferred that NAD synthesis may be impaired in disease, leading to accumulation of niacin/nicotinamide and low NAD, while TG lowered niacin/nicotinamide levels, possibly reflecting consumption for NAD synthesis. Pyridoxamine (PM) is part of vitamin B6 [35]. Vitamin B6 supports chondrogenic differentiation [36], and B6 deficiency can weaken collagen crosslinking and bone strength [37]. Vitamin B6 metabolism has been linked to PMOP progression [38]. In this study, PM was lower in OVX at week 8, while TG increased PM, suggesting a possible role in supporting collagen and bone strength.
During bone remodeling, osteoblasts mediate bone formation. ALP and ARS staining reflect early osteogenesis and late mineralization, respectively, and typically increase under osteogenic induction [39–40]. Here, TG enhanced ALP activity and mineralized nodules more effectively than TFE alone. BMPs are members of the TGF-β superfamily and play key roles in osteogenic differentiation. BMP-2 binds receptors, activates Smad1/5/8 (with Smad4), and promotes transcription factors such as RUNX2 and downstream OSX, ultimately promoting osteogenic protein synthesis, extracellular matrix deposition, and mineralization [41]. Previous research reported that TFE promotes BMSC osteogenesis via the BMP-2/RUNX2/OSX pathway [42]. This study focused on BMP-2/Smad signaling and found TG significantly increased BMP-2, p-Smad1/5, RUNX2, OSX, and ALP expression compared with TFE alone, supporting a stronger pro-osteogenic effect mediated through BMP-2/Smad.
In summary, in vivo and in vitro results indicate that TG effectively improves OVX-induced bone metabolic abnormalities. Mechanisms may involve regulation of amino acid, nucleotide, and vitamin metabolism and activation of BMP-2/Smad signaling to promote osteogenesis. This study provides theoretical support for TG in PMOP prevention and treatment.
5. Industry Perspective: Botanical Ingredient Trends for Postmenopausal Bone Health-From Mechanism-Oriented Research to Standardized Manufacturing
SEO focus: herbal extract for postmenopausal osteoporosis, botanical ingredients for bone health, osteogenesis BMP-2/Smad pathway, Epimedium flavonoids (icariin) extract, metabolomics-guided botanical product development.
5.1 Why mechanism-led "Herb Extract for Postmenopausal Osteoporosis" content matters in B2B
For brands, importers, and formulators building women's health and bone health portfolios, the market increasingly rewards botanical concepts that can be explained in clear scientific logic-for example:
multi-pathway metabolic modulation (metabolomics-supported), and
osteogenesis-related signaling pathways such as BMP-2/Smad.
Studies like the one translated above are valuable because they connect composition (TFE + CS/GLcN) → phenotypic outcomes (BMD, osteoclast reduction, micro-CT indices) → molecular/biochemical evidence (BMP-2/Smad, PINP/CTX-I) → metabolic networks (vitamin B pathways, amino acid and purine metabolism).
5.2 Botanical ingredient sourcing: standardization, traceability, and scalable extraction are the real differentiators
In B2B supply, "herb" is not a single specification-it's a supply chain. For finished-product companies, the key questions are:
Can the ingredient be standardized (e.g., marker compounds, fingerprints)?
Are there quality systems and manufacturing controls to support long-term supply?
Can the supplier provide documentation packages for regulatory and customer audits?
5.3 Where Cistanche fits (trend + manufacturing capability, not a substitute claim)
Alongside established bone-health botanicals (e.g., Epimedium flavonoids), Cistanche has gained attention in the broader TCM and functional botanical space because of its long history of use in tonic formulas and the industry's growing preference for single-herb extracts and standardized actives that can be integrated into modern supplement formats.
For buyers who are evaluating botanical partners, a manufacturing-oriented supplier profile becomes important-covering:
vertically integrated sourcing,
extraction and processing capacity,
and company background transparency.
About our manufacturing capability (reference source): the company profile and factory background can be reviewed on the official "About Us" page of the supplier site. ([xjcistanche.com])
Compliance note for B2B blogs: If your product positioning targets "postmenopausal osteoporosis," ensure your outward-facing language remains structure/function or nutritional support–oriented (depending on your market), and avoid disease-treatment claims unless supported and permitted by local regulation.
Learn more (supplier reference pages):
Company overview / factory background: ([xjcistanche.com])
Related botanical content hub (TCM recipes and ingredient storytelling): (your provided reference link can be inserted once you confirm the final URL path is correct)
References
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