Dracocephalum Moldavica Research Articles

Dracocephalum Moldavica L.-Derived carbon dots for effective periodontitis treatment via reprogramming energy metabolism in Macrophages

Dracocephalum Moldavica L. (DML), a Chinese traditional medicine rich in flavone compounds (e.g., tilianin), has shown antioxidant and anti-inflammatory potential for alleviating inflammatory disorders. However, the extraction of active ingredients of DML is complex, and these extracts usually act synergistically to treat inflammatory diseases. In this study, the carbon dots derived from DML (T-CDs) were rapidly prepared via the one-pot method and showed good performance in biocompatibility and free radical scavenging capacity. Then, the T-CDs were loaded into the hydrogel and applied to treat periodontitis. The in vivo experiments demonstrated that T-CDs could promote the inflammation resolution of and the alveolar bone reparation in the periodontitis model after periodontal injection. With further investigation, it could be found that T-CDs involved the alternative polarization of macrophage by rebalancing glycolysis and mitochondrial respiration. Overall, it was the first time DML had been used to prepare carbon dots with anti-inflammation. This study provided an effective drug candidate for the treatment of periodontitis and offered an in-depth understanding of the therapeutic mechanism of T-CDs in periodontitis through macrophage immunomodulation.

Tilianin suppresses NLRP3 inflammasome activation in myocardial ischemia/reperfusion injury via inhibition of TLR4/NF-κB and NEK7/NLRP3.

Tilianin, a flavonoid compound derived from Dracocephalum moldavica L., is recognized for its diverse biological functionalities, in particular alleviating myocardial ischemia-reperfusion injury (MIRI). There is ample evidence suggesting that the NLRP3 inflammasome has a significant impact on the development of MIRI. In this study, rats undergoing the ligation and subsequent release of the left anterior descending (LAD) coronary artery and H9c2 cardiomyocytes subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) were used to investigate the effects of tilianin on NLRP3 inflammasome and its anti-MIRI mechanisms. Upon reperfusion, the rats were intraperitoneally injected with tilianin at doses of 3, 10, 30mg/kg. H9c2 cells were treated with tilianin at concentrations of 10, 30, and 50μg/mL. Echocardiography, TTC staining and TUNEL staining demonstrated that tilianin remarkably improved cardiac function and mitigated myocardial damage in MIRI rats. Additionally, notable inflammatory response reduction by tilianin was evidenced by subsequent hematatoxylin-eosin (HE) staining, inflammatory cytokines assay, and quantitative proteomics. Further western blotting analysis and immunofluorescence staining showed tilianin decreased the levels of TLR4, p-NF-κB, NLRP3, and ASC in MIRI rats and H9c2 cells exposed to OGD/R, alongside a significant reduction in cleaved gasdermin D, mature IL-1β and IL-18. Molecular docking, cellular thermal shift assay (CETSA) and co-immunoprecipitation (co-IP) assay revealed that tilianin impeded the interaction between NLRP3 and NEK7. Taken together, tilianin protects cardiomyocytes from MIRI by suppressing NLRP3 inflammasome through the inhibition of the TLR4/NF-κB signaling pathway and the disruption of the NEK7/NLRP3 interface. These findings underscore the potential of tilianin as a promising therapeutic candidate for MIRI.

Effect of Dietary Moldavian Balm (Dracocephalum moldavica L.) on Growth Performance, Antioxidant Status, Immune Response, and Gene Expression of Common Carp (Cyprinus carpio)

Abstract Chemical compounds used to prevent and control fish disease often cause environmental hazards; thus, alternative approaches as new and effective strategies are needed. The current investigation was performed with the aim of exploring the effects of dietary Moldavian balm (MB, Dracocephalum moldavica L.) on the growth, immune parameters, and antioxidant status of common carp (Cyprinus carpio). Fish (n=300, w=3.80±0.02 g) in four groups in triplicates were supplemented with 0%, 0.5%, 1%, and 2% MB. After 42 days, it was found that feed supplements increased final weight (FW), weight gain (WG), and specific growth rate (SGR) and decreased the food conversion ratio (FCR) (P<0.05). In addition, fish supplemented with 2% MB significantly showed higher serum total protein (TP), alternative complement pathway (ACH50), and glutathione peroxidase (GPx) (P<0.05). The 0.5% MB-supplemented fish represented higher levels of LYZ, alkaline phosphatase (ALP), total Ig, and ACH50 in their skin mucus in comparison with the unsupplemented fish (P<0.05). The results also indicated that 2% MB resulted in a significantly higher expression level of intestinal tumor necrosis factor (TNF- α) (P<0.05); however, the level of LYZ, interleukin-1β (IL-1 β), and TLRs decreased in supplemented fish. CAT and SOD expressions were increased in 0.5% MB supplement. In conclusion, MB could be recommended as an efficient feed additive to boost common carp’s growth, immunity, and health status.

Antioxidant and Immunological Properties of Dracocephalum moldavica Extract in Penaeus vannamei

Objectives: The effects of different concentrations of Dracocephalum moldavica extract (DME) supplement on growth, survival, antioxidants, some biochemical parameters and innate immunity of White leg shrimp (Penaeus vannamei) juveniles investigated. Methods: For 56 days, shrimp (4.5-5 g) received diets that were enhanced with 0 (control), 50, 100, 200 mg DME/kg feed to attain apparent satiety. Results: The results revealed that feeding shrimp with DME significantly improved Feed efficiency and Feed conversion ratio with an optimum level of 200 mg/kg diet. Dietary DME supplementation had no significant effects on their carcass composition. Total albumin and protein higher (P > 0.05) in DME-fed particularly at 200 mg/kg treatment On the other hand, shrimp fed 200 mg DME/kg feed experienced significant decreases in aspartate and alanine aminotransferase levels. As dietary extract levels increased, there was a significant (P < 0.05) increase in SOD and CAT activities, as well as LYZ and PO levels, so that treatments with 100 and 200 mg DME/kg feed had the highest values, and Significant (P < 0.05) reductions in MDA and NO levels were observed at high dietary extract levels of 100 and 200 mg DME/kg. Conclusion: P. vannamei's growth, antioxidant capacity and innate immunity were boosted by diets that contained 200 mg DME/kg feed.

Comparison of Dracocephalum Moldavica Essential Oil with Chlorhexidine on Cariogenic Bacteria.

Dental caries and periodontal disease are the most common oral problems. Chemical antibacterial agents often have side effects; thus, researchers have long been in search for organic and herbal products to prevent dental caries and periodontal disease. The present study has aimed to assess the effects of Dracocephalum moldavica essential oil on Streptococcus mutans, Streptococcus salivarius, Streptococcus sobrinus, and Lactobacillus acidophilus compared to Chlorhexidine (CHX). In this in vitro study, the plants were collected from Zanjan Province, Iran. Analysis of the essential oil was carried out by gas chromatography/mass chromatography. Micro-broth dilution and disc diffusion methods were used for assessment of the antimicrobial activity, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were evaluated. The mean diameter of the growth inhibition zones in the well plate method for Dracocephalum moldavica showed that it had greater antimicrobial activity against Lactobacillus acidophilusthan others (p Value= 0.01). Furthermore, Dracocephalum moldavica had higher antimicrobial activity than CHX. The results of MIC and MBC showed that Dracocephalum moldavica had lower antibacterial activity than CHX. Dracocephalum moldavica essential oil demonstrated antibacterial properties against cariogenic bacteria. Given that other favorable properties of these essential oils are confirmed, they may be suitable for use as antibacterial agents in the formulation of oral healthcare products.

Effect of chemical, organic fertilization and storage time on the yield and quality of the dragon head (Dracocephalum moldavica L.) plant

Effect of chemical, organic fertilization and storage time on the yield and quality of the dragon head (Dracocephalum moldavica L.) plant

Dracocephalum moldavica L. Extract Ameliorates Hypertension Through Modulating the Interaction Between miRNAs and Gut Microbiota in 2K1C Rats

Aim: To investigate the therapeutic effects of Dracocephalum moldavica L. extract (DME) against hypertension and elucidate the underlying molecular mechanism. Method: A two-kidney one-clip (2K1C) model was established in male Wistar rats, and DME or saline was administered via oral gavage for 4 weeks. Subsequently, the blood pressure was monitored. Fecal microRNAs (miRNAs), gut microbes, and host metabolites were evaluated through miRNA sequencing, 16S rRNA gene sequencing, and serum metabolomics analysis, respectively. Results: DME administration attenuated 2K1C-induced hypertension in rats. Additionally, DME alleviated the dysregulation of miRNA expression and gut microbiota dysbiosis in 2K1C rats. The correlation analysis confirmed the association of the differentially expressed miRNAs with gut microbes. Furthermore, non-targeted metabolomics analysis revealed different metabolic profiles in the serum of 2K1C rats compared to control rats. Interestingly, the changes in those serum metabolites were partially attenuated by DME. The metabolite enrichment analysis demonstrated that the differentially expressed metabolites were mainly involved in three metabolic pathways (Arachidonic acid metabolism, TCA cycle, and Alanine, aspartate and glutamate metabolism). Conclusion: DME can be suggested as an effective intervention in the prevention of hypertension through modulating the miRNAs, gut microbiota, and host metabolites. Our findings may give new insights regarding the treatment of hypertension.

Selection of Reference Genes for Expression Normalization by RT-qPCR in Dracocephalum moldavica L.

Dracocephalum moldavica is widely used as an ornamental, medicine, and perfume in industry. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) is widely and accurately utilized for gene expression evaluations. Selecting optimal reference genes is essential for normalizing RT-qPCR results. However, the identification of suitable reference genes in D. moldavica has not been documented. A total of 12 reference genes in D. moldavica were identified by PEG6000 (15%) treatment under hypertonia conditions in different tissues (roots, stem, leaves, flower, seeds and sepal) and during three stages of flower development, then used to validate the expression stability. There were four algorithms (delta Ct, geNorm, NormFinder, and BestKeeper) used to analyze the stability. Finally, the RefFinder program was employed to evaluate the candidate reference genes' stability. The results showed that ACTIN, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and EF1α (elongation factor-1α) were stable reference genes under the PEG6000 treatment. Heat shock protein 70 (HSP70) was the most stable gene across different flower development stages. ADP-ribosylation factor (ARF) was the most stable gene in different tissues and total samples. This study provides reliable gene expression studies for future research in D. moldavica.

Hypolipidemic activity and mechanism of active components of Dracocephalum moldavica L. tea

Hypolipidemic activity and mechanism of active components of <em>Dracocephalum moldavica</em> L. tea

Rapid purification and scale-up of tilianin using counter-current chromatography with rectangular horizontal tubing

In counter-current chromatography (CCC), linear scale-up is an ideal amplification strategy. However, when transferring from analytical to predictable preparative processes with high throughput, linear scale-up would be challenging due to limitations imposed by differences in instrument parameters, such as gravitational forces, tubing cross-section area, tubing length, column volume and flow rate. Some effective scale-up strategies have been studied for different instrument parameters, but so far, these scale-up works have only been tested on standard circular (SC) tubing.The previous research of our group found that rectangular horizontal (RH) tubing can double the separation efficiency compared with conventional SC tubing, and has industrial production potential. This paper used the separation of tilianin from Dracocephalum moldavica L. as an example to demonstrate how to scale up the optimized process from analytical SC tubing to preparative RH tubing. After systematic optimization of solvent systems, sample concentration and flow rate on the analytical CCC, the optimized parameters obtained were successfully transferred to the preparative CCC. The results showed that a crude sample of 2.07 g was successfully separated using a solvent system of n-hexane - ethyl acetate - ethanol - water (1:4:1:5, v/v/v/v) in reversed phase mode, and the three consecutive separations produced a total of 380 mg tilianin in 75 min with high purities of 98.3%, as analyzed by HPLC. The total throughput achieved from the analytical to semi-preparative scale was improved by 138 times (from 12 mg/h to 1.66 g/h), while the column volume was increased by only 46.5 times (from 15.5 mL to 720 mL). This is the successful application of CCC for the separation and purification of tilianin. Given that SC tubing is the traditional configuration for CCC columns, this study is a necessary step to prove the applicability of RH tubing columns for routine use and potential large-scale industrial applications.

Graphical biplot assessment of pre-sowing procedures on the seeds of Dracocephalum moldavica L. employing diverse nanoparticles

Graphical biplot assessment of pre-sowing procedures on the seeds of Dracocephalum moldavica L. employing diverse nanoparticles

Effect of total flavonoids of Dracocephalum moldavica L. On neuroinflammation in Alzheimer's disease model amyloid-β (Aβ1-42)-peptide-induced astrocyte activation

ABSTRACT One of the main pathological features noted in Alzheimer’s disease (AD) is the presence of plagues of aggregated β-amyloid (Aβ1–42)-peptides. Excess deposition of amyloid-β oligomers (AβO) are known to promote neuroinflammation. Sequentially, following neuroinflammation astrocytes become activated with cellular characteristics to initiate activated astrocytes. The purpose of this study was to determine whether total flavonoids derived from Dracocephalum moldavica L. (TFDM) inhibited Aβ1–42-induced damage attributed to activated C8-D1A astrocytes. Western blotting and ELISA were used to determine the expression of glial fibrillary acidic protein (GFAP), and complement C3 to establish the activation status of astrocytes following induction from exposure to Aβ1–42. Data demonstrated that stimulation of C8-D1A astrocytes by treatment with 40 μM Aβ1–42 for 24 hr produced significant elevation in protein expression and protein levels of acidic protein (GFAP) and complement C3 accompanied by increased expression and levels of inflammatory cytokines. Treatment with TFDM or the clinically employed drug donepezil in AD therapy reduced production of inflammatory cytokines, and toxicity initiated following activation of C8-D1A astrocytes following exposure to Aβ1–42. Therefore, TFDM similar to donepezil inhibited inflammatory secretion in reactive astrocytes, suggesting that TFDM may be considered as a potential compound to be utilized in AD therapy.

A network pharmacology approach to decipher the mechanism of total flavonoids from Dracocephalum Moldavica L. in the treatment of cardiovascular diseases

Aim of the studyCardiovascular disease (CVD) seriously endangers human health and is characterized by high mortality and disability. The effectiveness of Dracocephalum moldavica L. in the treatment of CVD has been proven by clinical practice. However, the mechanism by which DML can treat CVD has not been systematically determined.Materials and methodsThe active compounds in DML were screened by literature mining and pharmacokinetic analysis. Cytoscape software was used to construct the target-disease interaction network of DML in the treatment of CVD. Gene ontology and signalling pathway enrichment analyses were performed. The key target pathway network of DML compounds was constructed and verified by pharmacological experiments in vitro. A hydrogen glucose deprivation/reoxygenation model was established in H9c2 cells using hypoxia and glucose deprivation for 9 h combined with reoxygenation for 2 h. The model simulated myocardial ischaemic reperfusion injury to investigate the effects of total flavonoids of Cymbidium on cell viability, myocardial injury markers, oxidative stress levels, and reactive oxygen radical levels. Western blot analysis was used to examine NOX-4, Bcl-2/Bax, and PGC-1α protein expression.ResultsTwenty-seven active components were screened, and 59 potential drug targets for the treatment of CVD were obtained. Through the compound-target interaction network and the target-disease interaction network, the key targets and key signalling pathways, such as NOX-4, Bcl-2/Bax and PGC-1α, were obtained. TFDM significantly decreased LDH and MDA levels and the production of ROS and increased SOD activity levels in the context of OGD/R injury. Further studies indicated that NOX-4 and Bax protein levels and the p-P38 MAPK/P38 MAPK andp-Erk1/2/Erk1/2 ratios were suppressed by TFDM. The protein expression of Bcl-2 and PGC-1α was increased by TFDM.ConclusionsOur results showed that DML had multicomponent, multitarget and multichannel characteristics in the treatment of CVD. The mechanism may be associated with the following signalling pathways: 1) the NOX-4/ROS/p38 MAPK signalling pathway, which inhibits inflammation and reactive oxygen species (ROS) production, and 2) the Bcl-2/Bax and AMPK/SIRT1/PGC-1α signalling pathways, which inhibit apoptosis.

INTRODUCTION OF DRACOCEPHALUM MOLDAVICA L. IN VITRO CULTURE

B a c k g r o u n d . Dracocephalum moldavica L. is widely used in folk medicine in many countries of the world due to its antispasmodic, antibacterial, analgesic, sedative and anti-inflammatory properties. D. moldavica essential oil is often used in aromatherapy and cosmetology. The use of biotechnological methods for obtaining secondary plant metabolites has become increasingly popular in recent years. On the one hand, they serve as a source of substances beneficial for pharmacology, while on the other hand, aseptic microclones of medicinal plants can be utilized as model organisms to explore methods to enhance the biosynthesis of specific target products. The in vitro use of plants for the production of biologically active substances is also a crucial and pertinent method for the conservation of natural biodiversity. The aim of this study was to determine the optimal conditions for introducing the medicinal plant Dracocephalum moldavica into in vitro culture. M e t h o d s. To introduce Dracocephalum moldavica plants into in vitro culture, the conditions for seed sterilization were carefully selected. The effectiveness of treatment with succinic acid in combination with various concentrations of sodium hypochlorite solution (20%, 50%, or 70%) was assessed to obtain aseptic material and ensure a high percentage of seed germination. The duration of seeds viability after storage for 1, 2, and 3 years was determined by testing seed germination in soil and in vitro. R e s u l t s . Seeds of D. moldavica have a relatively low germination rate when planted in soil and quickly lose their viability. In vitro cultivation can significantly enhance the germination of fresh seeds and allow for a more efficient use of two- and three-year-old seeds. The stimulating effect of succinic acid on the seed germination and seedling development of Dracocephalum moldavica has been observed, but it also activated the growth of fungal and bacterial infections. Therefore, it is advisable to use this stimulant only along with a more intensive sterilizing agent. C o n c l u s i o n s . The optimal method for introducing Dracocephalum moldavica from seeds into in vitro culture has been determined: pre-treatment of seeds with succinic acid followed by sterilization using 70% sodium hypochlorite.

Tilianin attenuates inflammasome activation in endothelial progenitor cells to mitigate myocardial ischemia-reperfusion injury.

Tilianin (TIL), a bioactive component derived from Dracocephalum Moldavica L., has been recognized for its anti-inflammatory properties. However, its effects on the Nlrp3 inflammasome within endothelial progenitor cells (EPCs) during myocardial ischemia-reperfusion injury (MIRI) remain unexplored. This study aimed to elucidate the role of TIL in modulating Nlrp3 inflammasome activation under MIRI conditions. A mouse model of MIRI was established to assess the therapeutic potential of TIL. EPCs treated with TIL at concentrations of 5, 10, and 20 μM were administered into the myocardium before reperfusion. Additionally, the cardioprotective effects of TIL were further examined by pre-treating EPCs with the compound before exposing them to hypoxia/reoxygenation (H/R) using cardiomyocyte supernatants. The impact on Nlrp3 inflammasome was assessed through western blotting, immunofluorescence, and ELISA. Our results showed that TIL concentration-dependently inhibited Nlrp3 inflammasome-related protein levels,and inhibited Asc oligomerization and Asc-Speck complex formation in EPCs, resulting in improved the migratory capacity and vascular structure formation of EPCs. In addition, TIL-treated EPCs significantly attenuated I/R injury and improved cardiac function. These results suggest that TIL ameliorates the inflammatory response in EPCs by suppressing Nlrp3 inflammasome activation, thereby facilitating neovascularization in the myocardium and conferring protection against MIRI. The study provides valuable insights into the potential of TIL as a therapeutic agent for cardiovascular diseases linked to ischemia-reperfusion injury.

Corrigendum: Tilianin extracted from Dracocephalum moldavica L. induces intrinsic apoptosis and drives inflammatory microenvironment response on pharyngeal squamous carcinoma cells via regulating TLR4 signaling pathways

Corrigendum: Tilianin extracted from Dracocephalum moldavica L. induces intrinsic apoptosis and drives inflammatory microenvironment response on pharyngeal squamous carcinoma cells via regulating TLR4 signaling pathways

Dosing of leonardite/struvite compounds as phosphorus fertilizers increased biomass and nutrient uptake in a calcareous soil

Non-renewable sources of phosphorus (P) are running out, so it is necessary to recover P from secondary recyclable materials. This study aims to recover P from domestic sewage sludge as Leo/MAP (Leonardite/magnesium ammonium phosphate) composites and use them as phosphorus fertilizer. The characteristics of Leo/MAP composites, solubility, and kinetic of P-release from them were evaluated. The properties of composites were determined by using XRF (X-ray fluorescence) spectroscopy, as well as structural analysis by using SEM.(Scanning electron microscopy) In addition, the effect of Leo/MAP composites was investigated on the responses of the Moldavian Balm (Dracocephalum Moldavica) under greenhouse conditions compared with traditional P-fertilization (TSP (Triple-superphosphates)) – and unfertilized check in terms of morphological characteristics, and concentration of nutrients (P, Mg, and N) in a calcareous soil. Microscopic images of manufactured composites showed they are free of PO (Parasitic ova) and the presence of FC (Fecal coliforms) in the samples was negative. The results showed that PRecovery (%) from domestic sewage sludge increased by application of leonardite, and the solubility and the dissolution rate of P, N, and Mg from composites in HCl (0.5 M) and citric acid (20 g/L) are more than that in water. The kinetic of P release from composites was better fitted the Korsmeyer-peppas equation (R2 = 0.96–0.99) in water, and to the first-order equation (R2 = 0.87–0.98) in citric acid and HCl solutions. The n value of the Korsmeyer-peppas model showed that the mechanism of P release from the composites is based on the Fickian-diffusion (n < 0.45). The highest positive impact of P-fertilization on morphological characteristics, and concentration of nutrients in Moldavian Balm plants were as follows: L75/MAP > L50/MAP > L25/MAP > L0/MAP. Therefore Leo/MAP composites, especially with a high amount of leonardite, are applicable to increase the growth of Moldavian Balm in a calcareous soil that is one of the important medical plants with its unique characteristic for human's health.

Characterization of dragonhead (Dracocephalum moldavica L.) landraces: Genetic, chemotypic, and agro-morphologic perspectives

Dragonhead (Dracocephalum moldavica L.), an endangered herb, faces significant threats due to the loss of biodiversity driven by land-use change. The seeds of seven landraces of D. moldavica were collected from various native populations in different regions of Northwest Iran. Subsequently, they were cultivated in 10 replications, spanning a total of 70 plots. This cultivation took place at the Ekbatan station of the Agriculture and Natural Resources Research Center in Hamadan province. This study aimed to assess the morphological traits, essential oil (EO) content, and genetic diversity within seven populations of Dracocephalum moldavica. Significant variations in morphological traits were observed among populations, indicating phenotypic diversity and potential for selective breeding programs. The EO content ranged from 0.03 % to 0.13 % (w/w), with the highest in the HO population and the lowest in the KE population. EO yield varied from 1.97 in the KE populations to 13.97 Kg/ha in the HO populations. The chemical composition of the EOs varied among populations and consisted of 18–24 components, with geranyl acetate being the dominant component. PCA analysis identified population differences primarily related to geranyl acetate, nerol, geraniol, neryl acetate, and neral. The observed variation in EO composition may result from genetic and environmental factors. ISSR analysis revealed high genetic diversity, with 84 out of 94 scorable bands being polymorphic. Cluster and PCoA analysis revealed distinct genetic clusters among populations, although some diversities did not align with geographical origin, suggesting seed transfer or high heterozygosity. The study highlights the effectiveness of ISSR markers in assessing genetic diversity and provides insights into the genetic structure and variability of D. moldavica populations. Furthermore, the correlation between ISSR markers and phytochemical traits suggests their potential for breeding programs to enhance important traits in D. moldavica. Considering both phytochemical and morphological properties is crucial in breeding programs due to pleiotropic effects of genetic loci.

Dracocephalum moldavica L. extract alleviates experimental colitis in rats by modulating gut microbiome and inflammatory pathways.

Several studies have revealed that an imbalance of the intestinal microbiota is involved in intestinal inflammation associated with ulcerative colitis (UC). Therefore, regulating the homeostasis of gut microbiota is critical for treating UC. Dracocephalum moldavica L. (DML) extract, a common traditional Chinese medicine, has been demonstrated to possess numerous pharmacological effects, such as antioxidative, anti‑inflammatory, and antibacterial properties. The aim of the present study was to evaluate the beneficial effects of DML extract and the probable mechanism of action in a dextran sulfate sodium‑induced chronic colitis model. It was found that DML extract ameliorated UC by improving disease activity index, weight loss, colon length, and histological scoring. DML extract administration also enhanced the count of Lactobacillus and reduced the count of Romboutsia. Furthermore, the results of network pharmacology analysis revealed that the active ingredients (including luteolin, rosmarinic acid, oleanolic acid, ursolic acid, apigenin, acacetin, kaempferol, and isorhamnetin) in the DML extract were closely associated with anti‑inflammatory activity via various signaling pathways, including the NF‑κB, IL‑17, TNF, and Toll‑like receptor (TLR) signaling pathways. Western blot analysis further indicated that DML extract downregulated the expression of members of the TLR4/NF‑κB signaling pathway, which was associated with colitis. Thus, it was hypothesized that DML extract exerted its anti‑colitis effects by modulating the gut microbiota and inflammatory pathways.

Foliar co-application of zinc oxide and copper oxide nanoparticles promotes phytochemicals and essential oil production in dragonhead (Dracocephalum moldavica)

Individual nanoparticle application has been documented to promote plant production; however, whether co-application of two nanoparticles (NPs) is more sustainable and significantly promotes plant production is unclear. Herein, foliar co-applications of two NPs or their conventional fertilizer forms on the growth, micronutrient (copper and zinc) enrichment, primary productivity, and essential oil (EO) production in a medicinal annual, dragonhead (Dracocephalum moldavica L.), were investigated. Treatments included 1:1 ratio of zinc oxide nanoparticles (ZnONPs):copper oxide nanoparticles (CuONPs) (40–400 mg/L), and compared with individual NPs, individual zinc suspension (ZnS) and chelated copper (chelated-Cu), and their combination, at equivalent concentrations. Results showed that the highest bioenrichment of Zn and Cu was observed with 80–160 mg/L ZnS+chelated-Cu, 400 mg/L ZnONPs+CuONPs, or ionic combination treatments. A dose-dependent increase in hydrogen peroxide and malondialdehyde was observed with co-treatment of NPs or ions, and oxidative stress responses were higher with NPs or ions co-treatment than individual treatment. With 160 mg/L ZnONPs+CuONPs treatment, total chlorophyll, aboveground biomass, and essential oil production increased significantly compared to control, 160 mg/L CuONPs, and 160 mg/L ZnONPs (227, 157 and 823 %; 58, 79, and 51 %; and 46, 80, and 3 %, respectively). Flavonoid and anthocyanin content also increased significantly (58 and 50 %, respectively) with ZnONPs+CuONPs compared to ZnS+chelated-Cu and were higher than ZnONPs or CuONPs alone by 10 and 25 %, and 37 and 36 %, respectively. More importantly, EO production and quality improved with 160 mg/L ZnONPs+CuONPs treatment compared to control. Taken together, our findings showed that foliar co-treatment of 160 mg/L ZnONPs+CuONPs could significantly improve primary productivity, aboveground biomass, and EO quality and yield in dragonhead grown in semi-arid field conditions; and thus, 160 mg/L ZnONPs+CuONPs is recommended as an optimal foliar co-treatment strategy for promoting sustainable plant production in semi-arid regions where soil nutrients and water are limiting factors inhibiting crop yield.