Salvia Miltiorrhiza Research Articles

Heterologous Expression of MYB Gene (Rosea1) or bHLH Gene (Delila) from Antirrhinum Increases the Phenolics Pools in Salvia miltiorrhiza

Phenolic acids have health-promoting properties, however, but their low concentrations in Salvia miltiorrhiza limit broader medicinal applications. MYB and bHLH transcription factors activate multiple target genes involved in phenylpropanoid metabolism, thereby enhancing the production of various secondary metabolites. We introduced the MYB transcription factor Antirrhinum Rosea1 (AmROS1) or Delila (AmDEL) into S. miltiorrhiza and observed that antioxidant activity in transgenic plants increased by 1.40 to 1.80-fold. The total content was significantly higher in transformants compared to the controls. Furthermore, heterologous expression of AmROS1 or AmDEL triggered moderate accumulations of rosmarinic acid and salvianolic acid at various growth stages. Levels of total phenolics, total flavonoids, and anthocyanins were significantly elevated. These biological and phytochemical alterations were correlated with the upregulated expression of genes involved in phenolic acid biosynthesis. Our findings demonstrate that AmROS1 and AmDEL function as a transcriptional activator in phenolic acids biosynthesis. This study offers further insights into the heterologous or homologous regulation of phenolics production, potentially enabling its engineering in S. miltiorrhiza.

A Spray-Dried Self-Stabilizing Nanocrystal Emulsion of Traditional Chinese Medicine: Preparation, Characterization and ex vivo Intestinal Absorption

AbstractSalvia miltiorrhizae (Danshen, the rhizome of Salvia miltiorrhiza Bge.) and Chuanxiong rhizome (Chuanxiong, the rhizome of Ligusticum chuanxiong Hort.) are two traditional Chinese medicines that have been widely used for the treatment of cardiovascular and cerebrovascular diseases. However, formulation development is difficult due to the complexity of the active ingredients, particularly the water-insoluble tanshinones and volatile oil of Chuanxiong rhizome, which cannot be absorbed via oral administration in conventional dosage forms. This study aimed to develop a self-stabilized nanocrystal emulsion co-loading the water-soluble, insoluble, and volatile active ingredients of Salvia miltiorrhizae and Chuanxiong rhizome to improve the bioavailability of the drugs. In this work, a high-pressure homogenization method was used to prepare a self-stabilizing nanocrystal emulsion. The emulsion was then spray-dried using hydroxypropyl-β-cyclodextrin. The dispersibility and storage stability of the spray-dried emulsion, the particle size and morphology of the emulsion droplets, and the drug content and phase distribution of the reconstituted emulsion were evaluated. An everted intestinal sac model was established, and high-performance liquid chromatography was used to determine the concentration of six active components (ferulic acid, salvianolic acid B, senkyunolide A, ligustilide, cryptotanshinone, and tanshinone IIA) and to assess the cumulative uptake amount of the drug and the apparent permeability coefficient. A mixture of the crude materials of tanshinones extract, total salvianolic acid, ferulic acid, and volatile oil of Ligusticum Chuanxiong was used as a control. The results showed that the spray-dried emulsion can be easily reconstituted into a uniform submicron emulsion with no significant changes in particle size, morphology, and microstructure of the emulsion droplets compared with the original emulsion before drying. The self-stabilizing nanocrystal emulsion significantly improved the intestinal absorption of water-insoluble components (tanshinone IIA, cryptotanshinone, and ferulic acid), and volatile oil components (senkyunolide A and ligustilide). Overall, the spray-dried self-stabilizing nanocrystal emulsion represents a potential oral formulation for Salvia miltiorrhiza and Chuanxiong rhizoma.

Salvianolic acid B improves DEN/CCl4/C2H5OH-induced renal fibrosis by regulating YAP/TAZ and pSmad3C/pSmad3L

Salvianolic acid B (Sal B) is a natural compound extracted from the root of Salvia miltiorrhiza, which has multiple biological activities. Targeting Hippo pathway transcriptional coactivators YAP/TAZ combined with Smad3 is an effective strategy against renal fibrosis. However, whether Sal B improves DEN/CCl4/C2H5OH-induced renal fibrosis and regulates YAP/TAZ and Smad3 phospho-isoform, pSmad3C/pSmad3L is unknown. Presently, we used DEN/CCl4/C2H5OH to induce renal fibrosis in mice and TGF-β1 or MST1/2 inhibitor XMU-MP-1 to induce HK-2 cells to investigate the activity and molecular mechanisms of Sal B. The results demonstrated that Sal B ameliorated DEN/CCl4/C2H5OH-induced renal fibrosis, including reducing renal tissue injury, improving renal function, and decreasing collagen synthesis. Furthermore, Sal B downregulated YAP/TAZ and promoted their phosphorylation in vivo and in vitro. Moreover, Sal B upregulated pSmad3C but downregulated pSmad3L in vivo and in vitro. Collectively, this study revealed that Sal B may improve DEN/CCl4/C2H5OH-induced renal fibrosis by regulating YAP/TAZ and pSmad3C/pSmad3L.

Structure-Guided Discovery of a Potent Inhibitor of the Ferric Citrate Binding Protein FecB in Vibrio Bacteria.

Infections caused by Gram-negative bacteria present a significant risk to human health worldwide. Novel strategies are needed to deal with the challenge caused by drug-resistant bacteria. Here, we report a new approach to combat infections by targeting iron-binding proteins to suppress bacterial growth. We investigated the function of the conserved periplasmic binding protein FecB from Vibrio alginolyticus. FecB was known to play a crucial role in the bacterial growth and to relate with biofilm formation. We then solved the crystal structures and elucidated the binding mechanism of FecB with ferric ion chelated by citrate. The results indicated that FecB binds weakly to one citrate molecule and strongly to the Fe3+-(citrate)2 complex. Based on these results, a structure-based virtual screening approach was conducted against FecB to identify small molecules that block ferric citrate uptake. Further evaluations in vivo and in vitro demonstrated that salvianolic acid C significantly suppressed bacterial growth, indicating that targeting bacterial nutrient absorption is a promising strategy for identifying potential antibacterial drugs.

Structure‐Guided Discovery of a Potent Inhibitor of the Ferric Citrate Binding Protein FecB in Vibrio Bacteria

AbstractInfections caused by Gram‐negative bacteria present a significant risk to human health worldwide. Novel strategies are needed to deal with the challenge caused by drug‐resistant bacteria. Here, we report a new approach to combat infections by targeting iron‐binding proteins to suppress bacterial growth. We investigated the function of the conserved periplasmic binding protein FecB from Vibrio alginolyticus. FecB was known to play a crucial role in the bacterial growth and to relate with biofilm formation. We then solved the crystal structures and elucidated the binding mechanism of FecB with ferric ion chelated by citrate. The results indicated that FecB binds weakly to one citrate molecule and strongly to the Fe3+‐(citrate)2 complex. Based on these results, a structure‐based virtual screening approach was conducted against FecB to identify small molecules that block ferric citrate uptake. Further evaluations in vivo and in vitro demonstrated that salvianolic acid C significantly suppressed bacterial growth, indicating that targeting bacterial nutrient absorption is a promising strategy for identifying potential antibacterial drugs.

Research Progress on the Pharmacological Mechanism of Traditional Chinese Medicine Danshen in the Treatment of Cerebral Ischemic Stroke

Cerebral ischemic stroke (CIS) is an acute cerebrovascular disease characterized by focal neurological deficits. Many modern scholars believe that the pathogenesis of CIS is related to factors such as endothelial injury, oxidative stress imbalance, neuroinflammation, cell apoptosis and autophagy, calcium overload, and excessive accumulation of oxygen free radicals. Traditional Chinese medicine believes that the pathogenesis of CIS is obstructed meridians, phlegm and blood stasis, and obstruction of the brain and orifices. Danshen, as a commonly used traditional Chinese medicine in clinical practice, has the effects of promoting blood circulation and removing blood stasis, unblocking meridians and relieving pain, clearing the heart and eliminating restlessness, cooling blood and eliminating carbuncle. In addition, modern pharmacological research has found that the specific pharmacological components of Salvia miltiorrhiza include tanshinone, salvianolic acid, shikimic acid, danshensu, protocatechualdehyde, etc. It has antioxidant stress, regulation of cell autophagy, anti-inflammatory factors, antiplatelet aggregation, and protection of vascular endothelium. This article aims to analyze and summarize the pharmacological mechanism of salvia miltiorrhiza in treating CIS, providing scientific basis for clinical application of salvia miltiorrhiza in treating CIS.

Biomass and Phenolic Acid Accumulation in Salvia austriaca Hairy Roots Grown in Temporary Immersion and Mist-Trickling Bioreactors

Transformed roots of Salvia austriaca were cultivated for 45 days in various systems, including Erlenmeyer flasks, a temporary immersion system (TIS) bioreactor, and a mist-trickling bioreactor, under controlled light conditions. The mist-trickling bioreactor yielded the highest biomass, with fresh and dry weights of 155.4 g/L and 10.2 g/L, respectively. Quantitative UHPLC analysis of hydromethanolic extracts revealed the biosynthesis of significant phenolic acids: caffeic acid, rosmarinic acid, and salvianolic acid A. Among these, rosmarinic acid was the most abundant, with its concentration varying based on the cultivation system. The highest total phenolic acid content, 165 mg/L, was obtained in the mist-trickling bioreactor, demonstrating its superiority in both biomass production and phenolic acid biosynthesis. This study highlights the potential of mist-trickling bioreactors for optimizing growth and metabolite production in S. austriaca transformed root cultures.

Salvianolic acid B improves the microcirculation in a mouse model of sepsis through a mechanism involving the platelet receptor CD226.

Salvianolic acid B (SalB) demonstrates diverse clinical applications, particularly in cardiovascular and cerebral protection. This study primarily investigated the effects of SalB on sepsis. The model of sepsis via caecal ligation puncture (CLP) was established in male C57BL/6 mice. Therapeutic effects of SalB on hepatic and pulmonary injury, inflammatory responses and microcirculatory disturbances in sepsis were evaluated. Platelet aggregation and adhesion were measured via flow cytometry and an adhesion test. After overexpression of platelet-related activating molecules by 293T cells, the efficient binding of SalB and platelet CD226 molecules was further evaluated. Finally, neutralizing antibody experiments were used to assess the mechanism of SalB in alleviating the progression of sepsis. SalB mitigated hepatic and pulmonary impairments, reduced inflammatory cytokine levels and enhanced mesenteric microvascular blood flow in septic mice. SalB enhanced CLP-induced reduction of platelet count and platelet pressure cumulative volume. SalB reduced platelet adhesion to endothelial cells and platelet aggregation to leukocytes. A high binding efficiency was observed between SalB and the platelet adhesion molecule CD226. Ex vivo, interactions between SalB and platelets from CD226-knockout mice were markedly decreased. In vivo administration of CD226 neutralizing antibodies significantly delayed disease progression and enhanced mesenteric microcirculation in septic mice. In our murine model of sepsis, treatment with SalB improved the microcirculatory disturbance and hindered the progression of sepsis by inhibiting platelet CD226 function. Our results suggest SalB is a promising therapeutic approach to the treatment of sepsis.

A derivative of tanshinone IIA and salviadione, 15a, inhibits inflammation and alleviates DSS-induced colitis in mice by direct binding and inhibition of RIPK2.

Inflammatory bowel diseases (IBDs) are chronic inflammatory conditions primarily affecting the gastrointestinal tract. Previous studies established the role of the NF-κB signaling pathway in the development of IBDs, suggesting that anti-inflammatory therapies might offer a viable treatment strategy. Tanshinone IIA and salviadione, both derived from Salviae Miltiorrhizae Radix et Rhizoma, possess anti-inflammatory and anti-oxidative activities. A series of new compounds were synthesized by hybridizing salviadione with tanshinone. Among these compounds, 15a showed beneficial effects in LPS-induced acute lung injury and diabetes-induced renal injury mouse models. The current study explored the therapeutic efficacy of 15a using both acute and chronic colitis models and elucidated the underlying mechanisms. DSS-induced colitis models were established in mice, where acute colitis was treated with compound 15a (5 or 10 mg·kg-1·d-1) for 8 days, while chronic colitis mice received compound 15a (5 or 10 mg·kg-1·d-1, i.g.) during 2.5% DSS administration. The 15a treatment significantly alleviated DSS-induced pathological and inflammatory damages in both acute and chronic colitis mouse models. In mouse intestinal epithelial cell line MODE-K, pretreatment with compound 15a (5 or 10 μM) significantly suppressed LPS + L18-MDP-induced inflammatory responses. The receptor-interacting serine/threonine kinase 2 (RIPK2) was identified as a direct binding target of compound 15a using microarrays and recombinant human proteins. Moreover, 15a could directly bind to and inhibit the phosphorylation of RIPK2, leading to the suppression of the NF-κB and MAPK signaling pathways. Furthermore, LEU153 and VAL32 were identified within the KD domain of RIPK2 as critical amino residues for the binding of 15a. Briefly, the current findings demonstrate that compound 15a holds promise as a therapeutic agent for managing acute and chronic colitis.

Salvianolic acids modulate lifespan and gut microbiota composition in amyloid-β-expressing Drosophila melanogaster.

Alzheimer's disease (AD), a form of neurodegenerative disorder characterized by the accumulation of amyloid-β (Aβ), hyperphosphorylated Tau, and neuroinflammation. The increasing population affected by AD urges for the development of effective treatments. The correlation between AD and gut microbiome remains underexplored, potentially providing a better understanding of the disease. Salvianolic acid A (Sal A) and salvianolic acid B (Sal B) are the active components extracted from Salvia miltiorrhiza (Danshen), and their antioxidant, anti-inflammation and Aβ inhibition activities were shown previously. In this study, these compounds were used to investigate their effects on Aβ toxicity, using Drosophila melanogaster expressing human Aβ42 as the model organism, by examining their lifespan and changes in gut bacterial communities. The study used two batches of flies, reared on food with or without methylparaben (MP) supplementation to evaluate the influence of MP on this animal model during pharmacological studies. MP is a common antimicrobial agent used in flies' food. The treatment of Sal A prolonged the lifespan of Aβ-expressing flies reared on MP-supplemented food significantly (P < 0.001), but not those without MP. The lifespan of Sal B-treated flies did not show a significant difference compared to untreated flies for both groups reared on food with and without MP. Sal A-treated flies in the presence of MP exhibited a lower abundance of Corynebacterium and Enterococcus than the untreated flies, while Lactiplantibacillus was the most dominant taxa. Urea cycle was predicted to be predominant in this group compared to the untreated group. The control group, Aβ-expressing flies treated with Sal A and Sal B on MP-supplemented food had improved lifespan compared to their respective groups reared on food without MP, while untreated Aβ-expressing flies was the exception. The gut microbiota composition of flies reared on MP-supplemented food was also significantly different from those without MP (P < 0.001).

Exploring the Antidiabetic Potential of Salvia officinalis Using Network Pharmacology, Molecular Docking and ADME/Drug-Likeness Predictions.

A combination of network pharmacology, molecular docking and ADME/drug-likeness predictions was employed to explore the potential of Salvia officinalis compounds to interact with key targets involved in the pathogenesis of T2DM. These were predicted using the SwissTargetPrediction, Similarity Ensemble Approach and BindingDB databases. Networks were constructed using the STRING online tool and Cytoscape (v.3.9.1) software. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis and molecular docking were performed using DAVID, SHINEGO 0.77 and MOE suite, respectively. ADME/drug-likeness parameters were computed using SwissADME and Molsoft L.L.C. The top-ranking targets were CTNNB1, JUN, ESR1, RELA, NR3C1, CREB1, PPARG, PTGS2, CYP3A4, MMP9, UGT2B7, CYP2C19, SLCO1B1, AR, CYP19A1, PARP1, CYP1A2, CYP1B1, HSD17B1, and GSK3B. Apigenin, caffeic acid, oleanolic acid, rosmarinic acid, hispidulin, and salvianolic acid B showed the highest degree of connections in the compound-target network. Gene enrichment analysis identified pathways involved in insulin resistance, adherens junctions, metabolic processes, IL-17, TNF-α, cAMP, relaxin, and AGE-RAGE in diabetic complications. Rosmarinic acid, caffeic acid, and salvianolic acid B showed the most promising interactions with PTGS2, DPP4, AMY1A, PTB1B, PPARG, GSK3B and RELA. Overall, this study enhances understanding of the antidiabetic activity of S. officinalis and provides further insights for future drug discovery purposes.

Enhanced Extraction of Tanshinones from Salvia miltiorrhiza Using Natural-Surfactant-Based Cloud Point Extraction

Salvia miltiorrhiza (SM) contains the tanshinones, a compound with various pharmacological effects, and has been extensively studied as a pharmaceutical material. However, conventional methods for extracting tanshinones face challenges such as environmental hazards and high cost. In this study, we aimed to effectively extract tanshinones from SM using cloud point extraction (CPE) with lecithin, a natural surfactant. By optimizing various extraction conditions including the solid-to-liquid ratio, lecithin concentration, NaCl concentration, pH, and equilibrium temperature, the optimal extraction efficiency was achieved using 20 mL of solvent per 1 g of sample, 3% lecithin (w/v), 2% NaCl (w/v), pH 6, and room temperature (25 ± 2 °C). The CPE method, which minimizes the use of organic solvent and is eco-friendly, demonstrated improvements in extraction efficiency, with a 4.55% increase for dihydrotanshinone I, 8.32% for cryptotanshinone, 15.77% for tanshinone I, and 6.81% for tanshinone IIA compared to the conventional water extraction method. These results suggest that CPE is a promising, environmentally friendly, and efficient approach for extracting hydrophobic components from pharmacologically active materials such as SM, with potential applications across various fields of natural product extraction.

Exploring the Potential Mechanisms of Danshen for the Treatment of Ulcerative Colitis based on Serum Pharmacochemistry, Gene Expression Profiling, and Network Pharmacology: Regulation of Cell Apoptosis and Inflammatory Response.

As a traditional Chinese medicine, Danshen shows potential efficacy for treating ulcerative colitis (UC). However, the bioactive components and mode of action were unclear. This paper uses a combination of network pharmacology, serum medicinal chemistry, and gene expression profiling to clarify its possible molecular mechanism of action and material basis. Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was utilized to analyze the herbal components and metabolites from the serum of Danshen-treated mice. Gene expression profiles were applied to construct a database of Danshen action targets. Then, active ingredient-target-biological functional module networks were constructed to analyze the mechanism of action. Molecular docking has further confirmed the possibility of its components to the targets. As a result, 193 common targets between 1684 Danshen-related DEGs and 1492 UC targets were determined as the potential targets for Danshen in treatment with UC. Serum pharmacochemistry and target prediction showed that 22 components in serum acted on 777 targets. Intersection with common targets yielded 46 core targets, and an active ingredienttarget- biological functional module network was constructed for analysis. Network prediction and molecular docking results showed that the main action modules were inflammatory response and cell apoptosis, which mainly acted on targets SRC, RELA, HSP90AA1, CTNNB1, STAT3, and CASP3. The main components of Danshen intervention in UC were predicted to include Catechol, 3,9-Dimethoxypterocarpan, 8-Prenylnaringenin, Isoferulic acid, Salvianolic acid C, and Danshensu. The present study provides a scientific foundation for further explicating the mechanisms of Danshen against UC.

Comparative profiling of the absorbed compounds and metabolites, and pharmacokinetic studies of Danshen-Chuanxiong herb pair in rat plasma and brain using liquid chromatography-tandem mass spectrometry

Danshen-Chuanxiong (DS-CX) was a classic herb pair commonly used to treat ischemic stroke. Nevertheless, the metabolic conversion and pharmacokinetic behavior of DS-CX in vivo remains unclear. This work aimed to reveal the in vivo metabolic behavior of DS-CX through establishing metabolic profiles and performing multicomponent pharmacokinetics analysis. The mass defect filtering (MDF) strategy integrated with UHPLC-QTOF-MS was firstly developed to characterize the metabolites of DS-CX in rats’ plasma and brain. Moreover, a sensitive UHPLC-QQQ-MS method was utilized to perform the comparative pharmacokinetic studies of major active ingredients of DS-CX in rats’ plasma. A total of 111 exogenous compounds (29 prototype compounds and 82 metabolites) were identified in rat biological samples. The major metabolic pathways were hydroxylation, methylation, deoxidation, dehydration, hydrogenation, demethylation, hydrolysis, decarboxylation and glucuronidation binding reactions. According to the results of metabolites profiling, sixteen active compounds (8 phenolic acids, 5 phthalides and 3 tanshinones) were selected as markers for further comparative pharmacokinetics study. Compared with the oral administration of DS or CX alone, the higher Cmax of salvianolic acid B, crytotanshinone and tanshinone IIA; the shorter Tmax of lithospermic acid, rosmarinic acid and tanshinone IIA; as well as the higher AUC0−∞ of ferulic acid, rosmarinic acid, salvianolic acid B, senkyunolide I and crytotanshinone, could be found after co-administration of DS-CX (P < 0.05). This study provided the overall knowledge of metabolites profiling of DS-CX in vivo, which would help to understand the effective material basis and promote the clinical application of DC-CX herb pair.

Development of a new paradigm model for deciphering action mechanism of Danhong injection using a combination of isothermal shift assay and database interrogation

BackgroundIdentification of active components of traditional Chinese Medicine (TCM) and their respective targets is important for understanding the mechanisms underlying TCM efficacy. However, there are still no effective technical methods to achieve this.MethodsHerein, we have established a method for rapidly identifying targets of a specific TCM and interrogating the targets with their corresponding active components based on Isothermal Shift Assay (iTSA) and database interrogation.ResultsWe optimized iTSA workflow and identified 110 targets for Danhong injection (DHI) which is used as an effective remedy for cardiovascular and cerebrovascular diseases. Moreover, we identified the targets of the nine major ingredients found in DHI. Database interrogation found that the potential targets for DHI, in which we verified that ADK as the target for salvianolic acid A and ALDH1B1 as the target for protocatechualdehyde in DHI, respectively.ConclusionOverall, we established a novel paradigm model for the identification of targets and their respective ingredients in DHI, which facilitates the discovery of drug candidates and targets for improving disease management and contributes to revealing the underlying mechanisms of TCM and fostering TCM development and modernization.Graphical

Salvia miltiorrhiza in osteoporosis: a review of its phytochemistry, traditional clinical uses and preclinical studies (2014-2024).

Osteoporosis becomes a global public health concern due to its rising prevalence and substantial impact on life quality. Salvia miltiorrhiza Bunge (Salviae Miltiorrhizae Radix et Rhizoma, SM) has been firstly recorded in Shen Nong's Herbal Classic, and is frequently prescribed in conjunction with other herbs for the management of osteoporosis. This systematic review aims to comprehensively analyze the recent advances of SM on osteoporosis in traditional Chinese clinical uses and preclinical investigations. Literature encompassing pertinent studies were systematically retrieved across multiple databases, including the PubMed, Web of Science, Chinese National Knowledge Infrastructure, Chinese VIP Database, and Chinese Biomedical Literature Database. Original investigations spanning from February 2014 to March 2024, including traditional Chinese medicine (TCM) clinical trials and preclinical studies, were employed to analyze the effects and actions of SM on osteoporosis. Thirty-eight TCM clinical trials were identified to employ SM in combination with other herbs for the management of primary and secondary osteoporosis. The overall efficacy was between 77% and 96.67%. Forty preclinical studies were identified to investigate the effects and actions of SM and/or its ingredients on osteoporosis. The anti-osteoporosis actions of this herb may be attributed to inhibit osteoclastogenesis/bone resorption and promote osteoblastogenesis/osteogenesis. The ethanol extracts and its ingredients (tanshinones) inhibit osteoclastogenesis/bone resorption by inhibiting the MAPK/NF-κB/NFATc1 signaling pathway and cathepsin K-induced collagen degradation. Both ethanol extracts (tanshinones) and water extracts (Sal B and tanshinol) contribute to osteoblastogenesis by promoting osteogenesis and angiogenesis via activation of the Wnt/β-catenin/VEGF and ERK/TAZ pathways, and eliminating ROS production targeting Nrf2/ARE/HO-1 pathway. In conclusions, SM may offer a novel strategy for osteoporosis management. Well-designed clinical trials are still needed to evaluate the actions of this herb and its ingredients on bone remodeling.

Identification of compounds from Origanum compactum and Origanum elongatum using HPLC/UV-ESI-MS and comparative analysis of their antioxidant, antimicrobial, anticoagulant, and antidiabetic properties

The aim was to assess the phytochemical composition, phenolic component levels, and biological properties of the flowering tops of Origanum compactum and Origanum elongatum. The study employed phytochemical assays, spectrophotometric techniques for quantitative analysis of polyphenols, flavonoids, and tannins, and compound identification using HPLC/UV-ESI-MS. The antimicrobial, antioxidant, anticoagulant, and antidiabetic properties were examined both in vitro and in vivo. The results showed that the O. compactum extract had significantly high levels of total polyphenols, measuring 47.368 mg gallic acid equivalents per gram, and flavonoids, measuring 14.839 mg quercetin equivalents per gram. The phytochemical examination of O. compactum revealed that lithospermic acid accounted for 36.82 % of the chemicals detected, followed by salvianolic acid C at 12.57 % and ros-marinic acid at 6.01 %. The main constituents of O. elongatum are salvianolic acid C (14.46 %), luteolin-3-O-glucuronide (13.51 %), salvianolic acid B (12.24 %), rosmarinic acid (7.83 %), and rutin (6.18 %). The results demonstrated different levels of effectiveness against the investigated microorganisms, with the extract from O. compactum exhibiting better activity, particularly against Gram-negative bacteria, certain yeasts, and the fungus Aspergillus niger. The aqueous extracts of both Origanum species demonstrate significant antioxidant activity. O. compactum has a higher total antioxidant capacity (IC50 of 35.083 μg/mL) compared to O. elongatum (IC50 of 77.080 μg/mL). However, O. elongatum has a higher reducing power (35.697 μg/mL) compared to O. compactum (42.563 μg/mL). In vivo evaluations revealed that the aqueous extracts of O. compactum and O. elongatum possess significant antihyperglycemic and anticoagulant properties. The extracts demonstrated a marked reduction in blood glucose levels during the oral glucose tolerance test (OGTT) in Wistar rats and effectively prolonged both prothrombin time (PT) and activated partial thromboplastin time (aPTT), highlighting their ability to inhibit coagulation pathways. Moreover, their comparable efficacy to standard antihyperglycemic medications and absence of severe toxicity, even at high doses, underscore their therapeutic potential for safe and effective treatment applications. Between the two species, O. compactum exhibited superior efficacy in key biological activities such as antioxidant, antimicrobial, and anticoagulant properties, making it a strong candidate for therapeutic applications. This study underscores the value of Origanum species as a rich source of bioactive compounds, offering significant potential in pharmaceuticals, nutraceuticals, and agri-food industries. The findings pave the way for further exploration of their diverse applications.

Synergistic potentiation of the anti-metastatic effect of a Ginseng-Salvia miltiorrhiza herbal pair and its biological ingredients via the suppression of CD62E-dependent neutrophil infiltration and NET formation.

The combination of the roots of ginseng and Salvia miltiorrhiza is an effective approach for treating metastatic cancer in patients with Qi stagnation and blood stasis patterns. However, the molecular mechanism underlying the combined use of ginseng and Salvia miltiorrhiza is unknown. This study unveils the pharmacological foundation of ginseng and Salvia miltiorrhiza by examining the involvement of neutrophils in the critical process of tumor hematogenous metastasis. Additionally, by employing a reverse pharmacology research model (effect-target-constituent), potential potent components were screened, and the dominant component formulations were determined. An experimental lung metastatic model was constructed to compare the antitumor effects of ginseng and Salvia miltiorrhiza. RNA sequencing was employed to identify pivotal biological events and key targets, while the detection of CD62E expression and neutrophil extracellular traps (NETs) release was used to screen for effective substances in ginseng and Salvia miltiorrhiza. In addition, a comprehensive array of in vitro and in vivo experiments was conducted to explore the underlying mechanisms and therapeutic significance. Compared with single-herb use, the use of ginseng or Salvia miltiorrhiza significantly reduced tumor metastasis, which was accompanied by reduced neutrophil infiltration into the lungs. Cryptotanshinone (CPT), an active constituent of Salvia miltiorrhiza, can inhibit neutrophil adhesion and recruitment to lung tissue by downregulating the expression of E-selectin (CD62E) in endothelial cells. Moreover, the ginseng -derived ginsenoside Rg1 mitigated the formation of NETs in lung tissues and reversed the protumor effects of NETs. We further explored the efficacy of combination therapy with Rg1 and CPT, which also reduced tumor metastasis in vivo. Ginseng and Salvia miltiorrhiza exhibited a mutual potentiation of the anti-metastatic effect by suppressing both early and late stages of neutrophil-initiated metastasis cascade. Rg1 and CPT represent the synergistic ingredients from ginseng and Salvia miltiorrhiza, respectively.

Serum untargeted metabolomics analysis of mice after myocardial infarction affected by qiliqiangxin capsule

Qiliqiangxin (QLQX) capsule consists of 11 herbs, namely Huang qi (astragalus membranaceus), Ren shen (ginseng), Fu zi (radix aconiti carmichaeli), Dan shen (salvia miltiorrhiza), Ting li zi (lepidium seed), Ze xie (rhizoma alismatis), Yu zhu (radix polygonati officinalis), Gui zhi (cassia twig), Hong hua (carthamus tinctorious), Xiang jia Pi (cortex periplocae), Chen Pi (pericarpium citri reticulatae), and it is a standardized commercial formula designed to address yang deficiency and to restore the balance of qi in the heart. QLQX is also known to invigorate the blood and promote the circulation of the blood and to promote the use of fluids to relieve water retention and edema, and can be used in cardiovascular diseases such as mild to moderate congestive heart failure resulting from coronary artery disease and hypertension. The further research on the effect of QLQX on cardiac function in mice after myocardial infarction was manipulated. QLQX was given to mice in myocardial infarction model by gavage with appropriate dosage and the samples were analyzed at the end of the animal experiments through the UHPLC-Q-Exactive LC-MS. The liquid mass spectrometry was used to collect and followed by further analysis of the corresponding metabolites and metabolic pathways using metabolomics analysis. As a result, 9 differential metabolites were identified, with 15 being up-regulated and 4 down-regulated following intervention with QLQX. Then the metabolic pathways by KEGG enrichment pathway bubble diagram was analyzed, and 4 metabolic pathways were obtained, and combined with the metabolites that had been screened and analyzed together, finally the two differential metabolites, 2,5-Dihydroxybenzenesulfonic Acid and o-Cresol sulfate were found. The Glycerophospholipid metabolism pathway was closely related to the remaining seven differential metabolites, and the pathway might be an important pathway related to the effects of QLQX on cardiac function in mice.

Inhibitory Effects of Wild Origanum elongatum Extracts on Fusarium oxysporum Mycelium Growth and Spores Germination: Evidence from in-vitro, in-planta, and in-silico experiments

Fusarium wilt, caused by Fusarium oxysporum, is one of the most devastating diseases to crops including potatoes. This fungus is widespread and dispersed via resistant spores. In this study, organic (methanol, hexane, and chloroform) and aqueous extracts of the aerial parts of wild Origanum elongatum were tested against mycelium and spores of F. oxysporum in both in-vitro (using the poisoned food technique) and in-planta assays. Further, the chemical composition of the extract was identified via HPLC-PDA-MS/MS. Then, molecular modeling was used to clarify the antifungal activity of identified biomolecules. The obtained results showed that organic and aqueous extracts showed significant antifungal and anti-sporulation activities with different effectiveness depending on the extract, concentration, and application test. In in-vitro assays of the aqueous extract, the highest antifungal activity was recorded at concentrations of 10mgmL-1, while the highest anti-sporulation activity (91.79%) was observed at 50mgmL-1. In the organic extracts, methanol was the most effective against fungus with 100% inhibition of mycelium at 5mgmL -1 and the highest anti-sporulation effect (73.58%) at 20mgmL-1. In in-planta assays, the methanolic extract at 20mgmL-1 achieved 0% disease severity, while the aqueous extract at 50mgmL-1 reduced severity to 7%, both significantly more effective than the untreated control. Therefore, methanol was the most effective and these biological properties are supported by a wide range of bioactive molecules, including 56 molecules compared to only 27 molecules in the aqueous extract. These include glycosides of apigenin and kaempferol and salvianolic acid b. However, more research is needed to clarify the inhibition action of the individual components and their synergetic actions in controlled and field conditions.