Mechanism Of Herba Cistanches-Epimedium Herb-Fortune's Drynaria Rhizome in The Treatment Of Osteoporosis Based On Network Pharmacology And Molecular Docking Ⅱ

Table 1 Active ingredients of Drynaria fortunei, Cistanche deserticola and Epimedium

Figure 1 Target intersection genes

Figure 3 Drug-ingredient-target-stroke target network

GO enrichment analysis

5 KEGG pathway analysis

注：A：MMP-9-Suchilactone;B:MMP-9-quercetin;C：MMP-9-lu－ teolin;D：MMP-9-kaempferol

Table 2 Ligand-receptor binding energy

3 Discussion

OP is a serious bone metabolism disorder, especially in women over 60 years old. Due to the influence of hormone levels, the incidence of OP is significantly increased. The characteristic change of OP is that the rate of bone synthesis by osteoblasts is less than that of osteoclasts. In addition, factors such as reduced calcium absorption, vitamin D metabolism disorders, and decreased renal function can also lead to OP[2]. At the same time, OP can lead to increased bone fragility and easy fractures.

The primary purpose of treating OP is to prevent fractures, and several drugs such as bisphosphonates, Denosumab, teriparatide, and abaloparatide are often used clinically. The therapeutic effect of treating OP by stimulating the synthesis of osteoblasts or inhibiting the action of osteoclasts is not very clear.

HIGH GLYCOSIDE ECHINACOSIDE CISTANCHE EXTRACT FOR SALE FOR 

obvious, and also has certain side effects. Traditional Chinese medicine is particularly important in treating OP. Chinese herbal medicines such as Drynariae, Epimedium, and Cistanche deserticola are regarded by many Chinese medicine practitioners as high-quality medicinal materials in the treatment of OP. The main components of Drynariae include flavonoids, terpenes, phenolic acids, etc., which have a good therapeutic effect on OP [3, 4]. Epimedium, as a herbaceous plant of the Berberidaceae family, is often used in traditional Chinese medicine to nourish the kidneys, and aphrodisiac, strengthen muscles and bones, and dispel rheumatism in combination with other drugs. In recent years, more and more studies have found that epimedium glycosides also have immunity-regulating and anti-inflammatory effects [5, 6]. Based on metabolomics

 TRADITIONAL CHINESE MEDICINE TO NOURISH THE KIDNEYS

Studies have shown that Epimedium has a therapeutic effect on OP [7]. The traditional Chinese medicine compound icariin, Anemarrhena saponin and ferulic acid can promote the expression of bone matrix and regulate UMR-106 through BMP and Wnt/β-catenin signaling pathways. Osteoblast osteogenesis[8].

As a succulent plant growing in northern my country, Cistanche deserticola grows in deserts and arid areas. It was first recorded in the Compendium of Materia Medica to treat kidney yang deficiency, soreness and weakness of the waist and knees, and is known as "desert ginseng". The main components of Cistanche deserticola include cistanche deserticola glycoside, eugenol glycoside, β-sitosterol, carotene, amino acids and polysaccharides. Studies have shown [9] that Cistanche deserticola total glycosides can activate GSK-3β to degrade β-catenin factors through the Wnt/β-catenin signaling pathway, thereby affecting the cell cycle, promoting cell apoptosis, limiting cell migration, and inhibiting liver cancer.

Wang F et al. [10] have shown that Cistanche deserticola total glycosides and polysaccharides can promote osteogenic bone formation in SAMP6 mice by activating the Wnt/β-catenin signaling pathway, improve bone microstructural damage, and treat OP. In this study stigmasterol, beta-sitosterol, kaempferol,

There are a total of 46 main ingredients including luteolin, Icariin, quercetin, chryseriol, etc. The main targets involved are AKT1, ERS2, MMP-9, AR, mTOR, etc. Studies have shown that Drynariae, Epimedium, and Cistanche deserticola compounds can treat OP through factors such as AKT, mTOR, etc. In order to further identify Drynariae, Epimedium, and Cistanche deserticola, Epimedium for OP metabolism mechanism. This study used GO enrichment and KEGG analysis, and the results showed that the molecules involved in the three drugs are multi-target. GO

The entries include typical inflammatory factors IL-2, mitogen-activated protein kinase MAPK, apoptosis factor CASPASE, etc. The expression of IL-2 plays an important role in the Wnt/β pathway of metabolic bone disease [11] .

At the same time, studies have shown that puerarin inhibits osteoclastogenesis by inhibiting the TRAF6/ROS-dependent MAPK/NF-κB signaling pathway, thereby alleviating osteoporosis in ovariectomized mice [12]. The above results all show that the traditional Chinese medicine Drynariae, epimedium, and Cistanche deserticola can treat OP by regulating inflammation, inhibiting bone cell apoptosis, and regulating metabolism. KEGG results show that the pathways involved in Drynariae, Epimedium, and Cistanche deserticola mainly include cancer pathways, estrogen signaling pathways, PI3K-Akt signaling pathways, and other pathways. In diseases related to bone metabolism, the PI3K-Akt pathway plays an important role as a metabolic hub. Studies have shown [13] that increasing mechanical load and red blood cell production can increase osteogenesis and angiogenesis leading to OP after menopause by regulating the PI3K-Akt signaling pathway.

 TRADITIONAL CHINESE MEDICINE TO NOURISH THE KIDNEYS

The traditional Chinese medicine compound Eucommia ulmoides, Cuscuta semen, and Drynariae rhizome extracts improve glucocorticoid-induced osteoporosis by inhibiting osteoclastogenesis through the PI3K/Akt pathway [14]. Epimedium can also regulate various diseases through PI3K/Akt [15]. Molecular docking results show that MMP-9 and suchilac-

Small molecules such as tone, quercetin, luteolin, and kaempferol have good binding energy. The latest research shows that MMP-9 is an important protein involved in bone metabolic diseases such as OP [16]. Research shows that velvet antler polysaccharide and peptide extracts stimulate MAKP and MMP-9 signaling.

pathway inhibits bone resorption in high turnover osteoporosis [17]. Ligands quercetin and VitE can regulate the total number of bone cells to a lesser extent and prevent osteoporosis by regulating the expression of autophagy and apoptosis proteins such as LC3, beclin1 and caspase 3 [18]. Luteolin has multi-target and multi-channel effects in the treatment of osteoporosis, can reduce bone loss in OVX rats, and can promote osteogenic differentiation of BMSCs by regulating the activity of the PI3KAkt signaling pathway19].

Based on molecular docking, it has been initially shown that the small molecule compounds contained in traditional Chinese medicine compound ingredients can treat and prevent OP through multiple targets and multiple pathways.

To sum up, suchilactone, quercetin, luteolin, kaempferol and other ingredients are the key components of Drynariae, Epimedium and Cistanche deserticola, and factors such as AKT1, ERS2, MMP-9, AR, mTOR and other factors are the key targets of important small molecules. point, possibly treated through its pathways

and prevent OP. The analysis through network pharmacology experiments provides a theoretical basis for further exploring the traditional Chinese medicine compound Drynariae, Epimedium, and Cistanche deserticola in the treatment of OP, but further elucidation of the specific metabolic mechanism remains to be experimentally verified.

References:

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[2] Wu D, Cline-Smith A, Shashkova E, et al. T-Cell Mediated Inflammation in Postmenopausal Osteoporosis [J]. Front Immunol, 2021, 12: 687551. [3] Ma Jiangtao, Wan Lei, Huang Hongxing. Exploration of the mechanism of action of Drynaria fortunei in the treatment of osteoporosis based on network pharmacology [J]. Chinese Journal of Osteoporosis, 2020, 26(4): 490-496.

[4] Yang Wenjing, Huang Jian, Wang Wei, et al. Exploration of the mechanism of action of Drynaria fortunei in the treatment of osteoporosis based on network pharmacology and in vitro cell experiments Study on the mechanism of action of the active ingredients of "Sanqi-Gu Sui Bu" medicine for the treatment of osteoporosis [J]. Chinese Journal of Traditional Chinese Medicine, 2023, 48(4): 1087-1097.

[5]Bi Z, Zhang W, Yan X. Anti-inflammatory and immunoregulatory effects of icariin and icaritin [J]. Biomed Pharmacother, 2022, 151: 113180.

[6] Dai Z. Study on the Protective Effect and Mechanism of the Rhizoma Drynariae -Epimedium Formula on Osteoarthritis in Rats[J]. Contrast Media Mol Imaging, 2022, 2022: 2869707.

[7] Zhao JF, Xu JY, Xu YE, et al. High-Throughput Metabolomics Method for Discovering Metabolic Biomarkers and Pathways to Reveal Effects and Molecular Mechanism of Ethanol Extract From Epimedium Against Osteoporosis [J]. Front Pharmacol, 2020,11:1318. [8]Li M, Zhang ND, Wang Y, et al. Coordinate regulatory osteogenesis effects of icariin, timosaponin B II and ferulic acid from traditional Chinese medicine formulas on UMR-106 osteoblastic cells and osteoblasts in neonatal rat calvaria cultures [J].J ,2016,185:120-131