Structure analysis of an acidic polysaccharide isolated from Epimedium koreanum Nakai and its improvement on liver oxidative stress injury in STZ-induced diabetic rats via NRF2/HO-1 pathways.

Multiple myeloma (MM) is a hematological malignancy for which novel therapeutic strategies are urgently needed. Epimedokoreanin B (EKB), an isoprenylated flavonoid compound derived from the medicinal plant Epimedium koreanum, has demonstrated promising antitumour activity. However, its effects on MM have not been previously investigated. This study explores the anti-MM activity and the molecular interaction mechanisms between EKB and G-quadruplexes (G4) through a combination of biological activity assessments and computer-aided methodologies. EKB exhibits potent cytotoxicity against the MM cell lines U266 and RPMI-8226, with IC 50 values of 5.28 μM and 6.81 μM, respectively. It induces apoptosis in a concentration-dependent manner and specifically stabilizes the G-quadruplex (G4) structures of oncogenes such as c-Myc, c-KIT, Bcl-2, and k-RAS, as confirmed by BG4 immunofluorescence staining and fluorescence resonance energy transfer (FRET) assays. Additionally, EKB significantly suppresses the mRNA and protein expression levels of these genes in myeloma cells. Computational studies, including molecular docking, molecular dynamics (MD) simulations, and MM/GBSA calculations, confirm the strong binding affinity and stabilizing effects of EKB on G4s, revealing a mechanism involving π-π stacking and hydrogen bonding. This discovery underscores the unique ability of EKB to increase the stability of G-quadruplex structures, which are critical for regulating gene expression and inhibiting cancer cell proliferation. This research highlights the therapeutic potential of EKB in targeting these specific molecular structures, thereby offering a more effective approach to managing MM.

IntroductionEpimedium koreanum is a traditional Chinese tonic medicine obtained from the wild and cultivation, and its pharmacodynamic composition differs depending on the artificial cultivation method. Rhizosphere microorganisms influence the growth and active component accumulation of medicinal plants; however, the detailed composition, diversity, and connections to soil properties and medicinal herb active components in E. koreanum remain under-researched.MethodsIllumina NovaSeq technology was used to study the differences in rhizosphere microbial diversity and composition and pharmacodynamic constituents among cultivation methods, including wild tending (WT), bionic cultivated in forest (FP), and simulated habitat cultivation (SC).ResultsCompared with estimates for WT, SC and FP resulted in higher contents of active components in E. koreanum. This cultivation method improves the soil environment by increasing the soil pH, AK, TK, and AP contents. pH, TK, and AP were key factors affecting the bacterial community, while TN, AN, and SOM had significant impacts on the fungal community. Further analyses indicated that the active components of E. koreanum are positively correlated with the abundance of microorganisms, such as Bacillus and Humicola. These microbial communities were significantly enriched in the rhizosphere of FP. In addition, the rhizospheres of FP and WT were enriched with microbial taxa related to plant stress resistance, indicating that different cultivation methods have differential regulatory effects on the plant rhizosphere environment. The proposed FP cultivation method focuses primarily on nitrogen reduction and phosphorus-potassium enhancement along with microbial regulation to synergistically improve both medicinal quality and ecological adaptability.ConclusionSoil physicochemical properties and enzymatic activity under different cultivation methods affect soil microbial diversity and composition, thereby impacting plant growth and the synthesis of key components. This work provides a theoretical basis for the scientific and effective cultivation of high-quality E. koreanum.

The genus Epimedium includes popular Chinese medicinal plants, and icariin and its precursor icaritin are the key bioactive components of Epimedium. Here, we identified flavonoid 8-dimethylallyltransferase (F8DT) in the authentic medicinal material of Epimedium koreanum, which is a key gene in the icariin biosynthesis pathway. This enzyme can catalyze the synthesis of 8-prenylkaempferol (8PK) from kaempferol. The catalytic ability of the rate-limiting enzyme EkF8DT was significantly improved by truncating its N-terminal intrinsically disordered regions (IDRs) and enhancing the flux of the mevalonate pathway. Icaritin was also successfully synthesized by introducing flavonoid 4'-O-methyltransferase into the Saccharomyces cerevisiae strain. Finally, the highest production of 8PK and icaritin (3.6 g/L and 172.0 mg/L, respectively) was obtained in the S. cerevisiae strain.

Natural flavonoids have been shown to have many pharmacological activities. Efficient and continuous enrichment of total flavonoids with high content and low mobile phase usage from complex natural products is greatly needed at the moment. In this study, a new continuous chromatography system (CCS) with 6 zones and the18-column dynamic tandem connection technique was developed and used to enrich total flavonoids from Epimedium koreanum Nakai (EKN). The 18 columns were divided into 6 zones, and the principle of adynamic series of three columns was adopted for each zone to achieve continuous automatic separation and enrichment of total flavonoids under the control of alogic control valve. The CCS separation conditions were established based on single-column chromatography anda theoretical calculation model of the CCS. By means of the self-designed device and method, 485.11±3.16 g of total flavonoids were isolated from 16.2 kg of EKN. It is worth noting that the total content of 18 types of flavonoids in the samples enriched by the CCS was increased from 2.84±0.07% to 88.29±0.22%, the total recovery rate was 92.20±0.38%, and theRSD of each flavonoid was less than 5.0%. Furthermore, compared with single-column chromatography filled with the same volume of chromatography filler, the entire process saved about 2/3 of the mobile phase usage. In summary, the developed device and method could efficiently and continuously enrich total flavonoids from EKN with high-content and low mobile phase usage and would have a wide application prospect in the separation and enrichment of natural products.

Icariin promotes skeletal muscle mitochondrial biogenesis by the AMPK-PPARδ-PGC-1α signaling pathway: A potential mechanism for beneficial muscle function homeostasis

Development of Electrochemical fingerprints for Quick sensing identification of Epimedium Koreanum Nakai

The untargeted metabolomics-based molecular networking approach combined with multivariate analysis, proves to be an effective strategy for distinguishing raw materials in herbal medicine according to specific criteria. It exhibits the correlations between chemical constituents and the geographical habitats of plants, providing a valuable tool for ensuring quality control in mass production within the industry. In this study, we conducted a comprehensive investigation of the chemical compositions of Epimedium koreanum Nakai and performed comparative analyses on four extracts collected from distinct regions in South Korea using untargeted metabolomics tools. Through the comprehensive use of UPLC-QTOF/MS analysis and advanced statistical techniques, we elucidated the chemical composition, leading to the identification of key chemical markers. Additionally, the molecular networking analysis revealed distinct clusters of flavonoids and phenolic acids, highlighting the influence of regional factors on the metabolite profiles. These findings offer a promising avenue for enhancing quality control and traceability in the herbal medicine industry, underscoring the important role of geographical variation in the chemical profiles of herbal products.

Epimedium koreanum is a perennial herb of the Berberidaceae family, which is a traditional tonic in Chinese medicine. Seed germination of E. koreanum is difficult. Dormancy is an intrinsic factor that affects seed germination. Elucidating the molecular mechanism of seed dormancy and the lifting process of E. koreanum is of great significance for the breeding, conservation, and utilization of E. koreanum. Previous studies have concluded that E. koreanum seed dormancy breakage requires warm-temperature stratification followed by low-temperature stratification treatments. Therefore, we performed transcriptome sequencing using freshly harvested, untreated seeds (NS), seeds that developed a cotyledonary embryo after 90 d of constant-temperature stratification at 15 °C (CS), and seeds that broke dormancy by 90 d of stratification at 15 °C and 60 d of stratification at 5 °C (ND) in order to find the responsive genes and regulatory genes that regulate dormancy. A total of 92,867 genes with differential expression were identified. GO enrichment analysis highlighted redox processes, as well as structural components of the nucleus and ribosomes. KEGG enrichment analysis revealed a significant enrichment of phytohormone signaling pathways, which play a crucial role in seed dormancy release. Additionally, protein–protein interactions (PPIs) were predicted with starch and sucrose metabolic pathways. This study introduces a novel concept for a more profound comprehension of the molecular regulatory mechanism of E. koreanum and lays a theoretical foundation for the screening of E. koreanum candidate genes.

Assessment for the influence of processing methods on chemical compositions and pharmacodynamics of Epimedii Folium and revealing the component transformation mechanism

Epimedium koreanum is a traditional Chinese tonic herb. Its main medicinal components are secondary metabolites such as flavonoids and flavonol glycosides, but the biosynthetic mechanism is still unclear. Moisture conditions are a key environmental factor affecting E. koreanum medicinal components during harvesting. Different stages of E. koreanum under natural conditions after rainfall were selected to study changes in physiological properties, herb quality, and transcriptome. Malondialdehyde (MDA) content increased significantly in the D3 stage after rainfall, and protective enzyme levels also rose. Additionally, the flavonol glycoside content was relatively high. We sequenced the transcriptomes of D1, D3, and D9 (R) and identified differentially expressed genes (DEGs) related to flavonoid synthesis. This analysis allowed us to predict the roadmap and key genes involved in flavonoid biosynthesis for E. koreanum. These results suggest that the E. koreanum quality can be enhanced by natural drought conditions in the soil after precipitation during harvest. The harvesting period of E. koreanum is optimal when soil moisture naturally dries to a relative water content of 26% after precipitation. These conditions help E. koreanum tolerate a certain level of water scarcity, resulting in increased expression of flavonoid-related genes and ultimately enhancing the quality of the herb.

Interaction of Icariin from Epimedium koreanum and Udenafil on the Relaxation of Rabbit Penile Corpus Cavernosum Tissue

Epimedium koreanum Nakai, a well-known traditional Chinese medicinal herb, has been widely used to treat osteoporosis and sexual dysfunction for thousands of years. However, due to the decreasing population of East Asian natural resources, yearly output of Epimedium crude herb has been in low supply year by year. In this study, an unusual variety of E. koreanum was discovered in Dunhua, Jilin Province, the northernmost area where this variety was found containing 6 individuals, with three branches that had 27 leaflets, which is much more than the typical leaflet number of 9. Firstly, the novel E. koreanum varety was identified using DNA barcodes. Then, 1171 differentially expressed genes (DEGs) were discovered through parallel RNA-seq analysis between the newly discovered variety and wild type (WT) E. koreanum plant. Furthermore, the results of bioinformatics investigation revealed that 914 positively and 619 negatively correlated genes associated with the number of leaflets. Additionally, based on RNA-Seq and qRT-PCR analysis, two homologous hub TCP genes, which were commonly implicated in plant leaf development, and shown to be up regulated and down regulated in the discovered newly variety, respectively. Thus, our study discovered a novel wild resource for leaf yield rewarding medicinal Epimedium plant breeding, provided insights into the relationship between plant compound leaf formation and gene expression of TCPs transcription factors and other gene candidates, providing bases for creating high yield cultivated Epimedium variety by using further molecular selection and breeding techniques in the future.

Bioactive metabolites reveal the therapeutic consistency of epimedii folium from multi-plant sources for the treatment of kidney-yang deficiency

PurposeThis study aims to explore the effects of two endophytes of Epimedium koreanum Nakai on host plant growth and development and medicinal material quality and to promote the growth and development of ingredients derived from Epimedium koreanum Nakai.MethodsThe effects on the biomass, photosynthesis parameters, chlorophyll fluorescence parameters, activity of key enzymes, and active components in the process of growth and development in Korea were examined.ResultsFrom the analysis of biomass and photosynthesis data, both the S3 (Diaporthe cotoneastri) and S10 (Ilyonectria cyclaminicola) strains promoted the growth of Epimedium koreanum Nakai. Redundancy analysis (RDA) showed that the activity of S3 and the content of epimedin A were negatively correlated and that the peroxidase (POD) activity was positively correlated with the total flavonoid content.ConclusionThe application of the S3 (Diaporthe cotoneastri) and S10 (Ilyonectria cyclaminicola) strains can significantly increase the biomass and contents of total flavonoids, icariin, epimedin B, and epimedin C, but not epimedin A, in Epimedium koreanum Nakai host plants grown in field conditions. The S10 strain promotes faster growth of Epimedium koreanum Nakai than the S3 strain, and flavonoid 3-hydroxylase (F3H) catalyzes the synthesis of epimedin A and icariin by different pathways using the same substrate. POD plays a key role in total flavonoid synthesis in Epimedium koreanum Nakai.

Two new prenylated flavonol glycosides, namely Desmethylicaritin-3-O-β-D-glucopyranosyl-(1→3)-α-L(4″-O-acetyl) rhamnopyranosyl-7-O-β-D(6′′′′-O-acetyl)-glucopyranoside (1) and 5,7,3′,4′-tetrahydroxyl-8-(3,3-dimethylallyl)-flavonol-3-O-α-L-rhamnopyranoside (2), and one with no NMR spectral data reported (3) were isolated from Epimedium koreanum Nakai. Their structures were elucidated by 1D, 2D NMR and HRESIMS analysis. The identification of the sugar moieties was carried out by means of acid hydrolysis and HPLC analysis of their derivatives. The anti-pulmonary fibrosis activities result showed that compound 2 had significant inhibitory effects on A549 cell fibrosis, which was similar to that of the positive control drug, pirfenidone.

The aim of this study was to develop a topically applied formulation with the potential to alleviate arthritis ailments. A combination of two active ingredients, icariin from Epimedium L. (Species: Epimedium Koreanum) extract as a potential promoter of chondrogenesis and glucosamine sulfate as a precursor of cartilage tissues, was tested. In permeation studies, the potential for skin permeation of both substances was confirmed; however, the in vitro release test did not accurately reflect the degree of skin permeation. The in vitro release of icariin was at a level of 15.0–19.0% for the plant-extract-derived icariin and 29.0–35.0% for the pure substance. The level of glucosamine sulfate release was 38.4% (on average). For icariin of both origins, the release results were higher than those obtained via oral administration (about 12.0%), which shows the potential superiority of topical application. In addition, the physicochemical parameters that affect the in vitro release and performance of topical formulations were addressed. This preliminary research and permeation analysis of the formulation produced a promising picture of its prospects regarding arthritis treatment, although further investigation is needed.

BackgroundRhizosphere fungi and endophytic fungi play key roles in plant growth and development; however, their role in the growth of Epimedium koreanum Nakai at different stages remains unclear. Here, we used the Illumina MiSeq system, a high-throughput sequencing technology, to study the endophytic fungi and rhizosphere microbiome of Korean Epimedium.ResultsEpimedium koreanum Nakai rhizosphere soil and leaves had highly diverse fungal communities during the growth process. The relative abundance of soil fungi in the rhizosphere stage was higher than that of leaf endophytic fungi in the early growth stage, but the overall abundance was basically equal. Sebacina is a significantly divergent fungal genera, and Sebacina sp. are present among leaf fungi species in the rhizosphere soil of Epimedium koreanum Nakai. Sebacina sp. can move to each other in rhizosphere soil fungi and leaf endophytes. VIF (variance inflation factor) analysis showed that soluble salt, whole nitrogen, alkaline lysis nitrogen, whole phosphorus, total potassium, and fast-acting potassium are useful environmental factors for rhizosphere soil and leaf endophytic fungi: potassium, total nitrogen, whole phosphorus, and three environmental factors were significantly and positively associated with the relative abundance of Sebacina sp.Conclusions(1) This study is the first to clarify the species diversity of fungi in Epimedium koreanum Nakai leaf and rhizosphere soil. (2) Different fungal communities of rhizosphere soil fungi and leaf endophytic fungi at different growth stages of Epimedium koreanum Nakai were examined. (3) Sebacina sp. can move to each other between rhizosphere soil fungi and leaf endophytic fungi. (4) Nitrogen, phosphorus and potassium elements in the environment have a significant positive effect on the relative abundance of Sebacina sp.

Epimedokoreanin B inhibits the growth of lung cancer cells through endoplasmic reticulum stress-mediated paraptosis accompanied by autophagosome accumulation

Epimedii Folium is widely used worldwide as an herbal supplement, and the risk of its induced liver damage has emerged in recent years. Our preliminary study has found that, among several Epimedii Folium species specified in the Chinese Pharmacopoeia, Epimedium koreanum Nakai has a more severe propensity for hepatotoxicity. However, the mechanism of hepatotoxicity of Epimedium koreanum Nakai is still unclear. In this study, untargeted metabolomics was performed to analyze the serum and liver tissue to explore the mechanism of hepatotoxicity of Epimedium koreanum Nakai. The results of experiments in vivo showed that, after 28 days of exposure to Epimedium koreanum Nakai ethanol extract (EEE), the liver weight, levels of AST, ALP, TBIL, etc. in serum of rats in the EEE group were significantly increased, as well as severe cytoplasmic vacuolation appeared in the liver tissue, which suggested that EEE has significant hepatotoxicity. Subsequently, the results of metabolomics revealed significant changes in the metabolic profile in the liver and serum of rats after EEE exposure, in which metabolites in serum such as flavin mononucleotide, phenylacetylglycine, glutathione, l-tryptophan, and sphingomyelin were able to accurately identify liver injury caused by EEE and could be used as serum markers to reflect EEE-induced liver injury. The KEGG pathway enrichment analysis revealed that EEE caused extensive effects on rats' metabolic pathways. Some of the most affected pathways included glutathione metabolism, glutamate metabolism pathway, primary bile acid biosynthesis pathway, and sphingolipid metabolism pathway, which were all directed to the biological process of ferroptosis. Then, the main markers related to ferroptosis in the liver were examined, and the results demonstrated that the content of malondialdehyde was significantly increased, the activity of superoxide dismutase was significantly reduced, the ferroptosis inhibitory proteins GPX4 and System xc − were significantly downregulated, and the ferroptosis-promoting protein ACSL4 was significantly up-regulated. Judging from these results, we concluded that the mechanism of hepatotoxicity of Epimedium koreanum Nakai was probably related to the induction of ferroptosis in hepatocytes.