Antifungal Activity of Cadinane-Type Sesquiterpenes from Eupatorium adenophorum against Wood-Decaying Fungi.

The present study aimed to evaluate the antifungal activities of Eupatorium adenophorum against four strains of wood-decaying fungi, including Inonotus hispida, Inonotus obliquus, and Inonotus cuticularis. Bioguided isolation of the methanol extract of E. adenophorum by silica gel column chromatography and high-performance liquid chromatography afforded six cadinane-type sesquiterpenes. Their structures were identified by nuclear magnetic resonance and MS analyses. According to the antifungal results, the inhibition rate of the compound was between 59.85 % and 77.98 % at a concentration of 200 µg/mL. The EC50 values ranged from 74.5 to 187.4 µg/mL.

Green synthesis of silver nanoparticles using Eupatorium adenophorum leaf extract: characterizations, antioxidant, antibacterial and photocatalytic activities.

The green synthesis of metallic nanoparticles has tremendous impacts in various fields as found in recent years due to their low cost, easy and environmentally friendly synthesis. In this article, we report a simple and eco-friendly method for the synthesis of silver nanoparticles (AgNPs) using an aqueous Eupatorium adenophorum (E. adenophorum) leaf extract as a bioreductant. Interestingly, Fourier transform infrared (FTIR) spectroscopy analysis established that the E. adenophorum extract not only served as a bioreductant but also acted as a capping agent to stabilize the nanoparticles by functionalizing the surfaces. Various characterization techniques were adopted, such as X-ray powder diffraction (XRD), FTIR, ultraviolet-visible absorption (UV-Vis) spectroscopy, dynamic light scattering, scanning electron microscopy and energy-dispersive X-ray spectroscopy (EDX) to analyze the biosynthesized AgNPs. Biosynthesized nanoparticles were also explored for antioxidant, antibacterial and photocatalytic activities. The AgNPs showed improved free radical scavenging activity (IC50 48.96 ± 0.84 µg/mL) and bacterial inhibitory effects against both gram-positive (Staphylococcus aureus; 64.5 µg/mL) and gram-negative (Escherichia coli; 82.5 µg/mL) bacteria. Photocatalytic investigation showed AgNPs were effective at degrading rhodamine dye (78.69% in 90 min) when exposed to sunlight. These findings collectively suggest that E. adenophorum AgNPs were successfully prepared without the involvement of any hazardous chemical and it may be an effective antibacterial, antioxidant and promising agent for the removal of hazardous dye from waste water produced by industrial dyeing processes.

Characterization of defensive cadinenes and a novel sesquiterpene synthase responsible for their biosynthesis from the invasive Eupatorium adenophorum.

Eupatorium adenophorum is a malignant invasive plant possessing extraordinary defense potency, but its chemical weaponry and formation mechanism have not yet been extensively investigated. We identified six cadinene sesquiterpenes, including two volatiles (amorpha-4,7(11)-diene and (-)-amorph-4-en-7-ol) and four nonvolatiles (9-oxo-10,11-dehydroageraphorone, muurol-4-en-3,8-dione, 9-oxo-ageraphorone and 9ß-hydroxy-ageraphorone), as the major constitutive and inducible chemicals of E. adenophorum. All cadinenes showed potent antifeedant activity against a generalist insect Spodoptera exigua, indicating that they have significant defensive roles. We cloned and functionally characterized a sesquiterpene synthase from E. adenophorum (EaTPS1), catalyzing the conversion of farnesyl diphosphate to amorpha-4,7(11)-diene and (-)-amorph-4-en-7-ol, which were purified from engineered Escherichia coli and identified by extensive nuclear magnetic resonance (NMR) spectroscopy. EaTPS1 was highly expressed in the aboveground organs, which was congruent with the dominant distribution of cadinenes, suggesting that EaTPS1 is likely involved in cadinene biosynthesis. Mechanical wounding and methyl jasmonate negatively regulated EaTPS1 expression but caused the release of amorpha-4,7(11)-diene and (-)-amorph-4-en-7-ol. Nicotiana benthamiana transiently expressing EaTPS1 also produced amorpha-4,7(11)-diene and (-)-amorph-4-en-7-ol, and showed enhanced defense function. The findings presented here uncover the role and formation of the chemical defense mechanism of E. adenophorum - which probably contributes to the invasive success of this plant - and provide a tool for manipulating the biosynthesis of biologically active cadinene natural products.

An efficient, green, and easy-to-scale-up strategy for target-oriented isolating cadinene sesquiterpenoids from Eupatorium adenophorum Spreng.

A green and efficient strategy was established and optimized for target-oriented extraction, enrichment and separation of cadinene sesquiterpenoids from Eupatorium adenophorum Spreng., using the combination of supercritical fluid extraction, molecular distillation, and industrial preparative chromatography for the first time. The extraction conditions of supercritical fluid extraction were initially optimized by orthogonal experimental design. Under the optimum conditions, the contents of 9-oxo-10,11-dehydroageraphorone and 10Hß-9-oxo-ageraphorone, which were 55.00% and 6.01%, respectively, were much higher than conventional extraction methods. Then, the molecular distillation enrichment method was established and investigated by response surface methodology technology, which showed strong specificity for enriching target compounds and removing impurities from crude extracts. Under the optimum conditions of molecular distillation, total contents of cadinene sesquiterpenoids were increased to 89.19%. Finally, a total of 146 mg of 9-oxo-10,11-dehydroageraphorone and 29 mg of 10Hß-9-oxo-ageraphorone were easily obtained by industrial preparative chromatography, from 200 mg of distillation fraction, with purities over 99%. The contents of target components were analyzed by HPLC, and structures of them were identified by high-resolution MS, 1 H-NMR, and 13 C-NMR spectroscopy. These results indicate that it is a simple, effective, and eco-friendly strategy, which is easily converted into industrial scale.

Inhibitory activities of flavonoids from Eupatorium adenophorum against acetylcholinesterase.

Fifteen flavonoids isolated from the Eupatorium adenophorum showed inhibitory activities against acetylcholinesterase (AChE) isolated from Caenorhabditis elegans and Spodoptera litura. Their IC50 values ranged from 12.54 to 89.06µg/mL and 12.08 to 86.01µg/mL, respectively against the AChE isolated from the nematode and insect species. AChE was inhibited in a dose-dependent manner by all tested flavonoids, The isolated compound quercetagetin-7-O-(6-O-caffeoyl-ß-D-glucopyranoside) displayed the highest inhibitory effect against AChE from C. elegans and S. litura, with IC50 values of 12.54 µg/mL and 12.58 µg/mL, respectively. The structure-activity relationship of flavonoids on the inhibitory activities indicated that additional phenolic hydroxyl groups in the glucose were favorable for their inhibitory effects and the degree of increase in inhibitory activity also depended on the number of phenolic hydroxyl groups. The Lineweaver-Burk and Dixon plots indicated that quercetagetin-7-O-(6-O-caffeoyl-ß-d-glucopyranoside) is a reversible inhibitor against AChE. Quercetagetin-7-O-(6-O-caffeoyl-ß-d-glucopyranoside), 5,4'-Dihydroxytlavone and quercetin-3-O-ß-d-glucopyranoside inhibited AChE in a mixed-type competitive manner and these compounds might be the dual binding site AChE inhibitors. Further, nine compounds showed poisonous effects against C. elegans and inhibitory effects on the growth and development of S. litura.

Effects of oil extracts of Eupatorium adenophorum on Phytophthora capsici and other plant pathogenic fungi in vitro.

The antifungal activity of oils extracted from Eupatorium adenophorum was tested against five phytopathogens in vitro. Oil extracts inhibited the mycelial growth of Phytophthora capsici which causes phytophthora blight in pepper. The minimum inhibitory concentration of oils against P. capsici was 500µg/ml after 7days incubation. At the ultrastructural level, oil extracts caused complete disorganization of intracellular organelles, cytoplasm depletion, disruption of cytoplasmic membranes and the cell wall. Membrane permeability increased with the increasing concentration of oil extracts. These results suggested that these oil extracts exhibited multiple modes of action including disruption of the cell membrane system. Furthermore, oil extracts combined with synthetic fungicides synergistically inhibited mycelial growth of P. capsici, which creates the possibility of reducing fungicide concentration needed to successfully control phytophthora blight in commercial pepper production. This study's use of multiple methods of analysis has increased our understanding of the mode of action of E. adenophorum oil extracts against P. capsici.

Preparation, Phytochemical Investigation, and Safety Evaluation of Chlorogenic Acid Products from Eupatorium adenophorum.

Eupatorium adenophorum is widely distributed throughout the world's tropical and temperate regions. It has become a harmful weed of crops and natural environments. Its leaves contain bioactive compounds such as chlorogenic acid and may be used as feed additives. In this study, chlorogenic acid was extracted and separated from leaves of E. adenophorum. Three chlorogenic acid products were prepared with different purities of 6.11%, 22.17%, and 96.03%. Phytochemical analysis demonstrated that the main toxins of sesquiterpenes were almost completely removed in sample preparation procedure. The three products were evaluated for safety via in vitro and in vivo toxicological studies. All the products exhibited no cytotoxic effects at a dose of 400 µg/mL in an in vitro cell viability assay. When administered in vivo at a single dose up to 1.5 g/kg bw, all three products caused no signs or symptoms of toxicity in mice. These results encourage further exploration of extracts from E. adenophorum in feed additive application.

New chlamydosporol derivatives from the endophytic fungus Fusarium sp. #001.

Two new chlamydosporol derivatives, fusarilactone A (1) and fusarilactone B (2), together with nine known compounds (3-11), have been isolated from the crude extract of endophytic fungus Fusarium sp. #001. The structures of new compounds 1 and 2 were elucidated on the basis of extensive spectroscopic methods. Compound 1 showed mild cytotoxicities against three tumor cell lines (SMMC-7721, A-549, and MCF-7).

Evaluation acaricidal efficacy of botanical extract from Eupatorium adenophorum against the hard tick Haemaphysalis longicornis (Acari: Ixodidae).

The aim of this study was to evaluate the acaricidal activity of a botanical extract from Eupatorium adenophorum against the hard tick Haemaphysalis longicornis. This could result in developing effective extracts of E. adenophorum as a source of natural, low-toxicity plant-based acaricidal drugs. Adult engorged females of H. longicornis were collected from naturally infected goats. The engorged females were reared in the laboratory and their offspring (larvae and nymphs) were used as test ectoparasites. The toxic effects of botanical extracts from E. adenophorum against larvae and nymphs of H. longicornis were evaluated. The results showed that the extracts with 1.5 and 1.0g/ml (w/v) concentrations were toxic for H. longicornis, comparable to a toxic effect of 2% chlorpyrifos (positive control). The median lethal time (LT50) for larval and nymphal ticks with 1.5g/ml (w/v) concentration of extract were 0.790 (LT99=1.065) and 1.018 (LT99=10.608) hours, respectively, whereas the LT50 of 1.0g/ml (w/v) concentration were 1.445 (LT99=6.047) and 1.313 (LT99=29.932) hours for larval and nymphal ticks, respectively. At a concentration of 1.5g/ml (w/v), an acaricidal effect of 100% was achieved for both larval and nymphal ticks, while a concentration of 1.0g/ml (w/v) resulted in 100% (for larvae) and 93% (for nymphs) within a 6h period. In additional, we found that the relatively low concentration (0.5g/ml) also obtained a good acaricidal effect during the short experimental period, with 2.22 and 2.651h LT50 for larval and nymphal ticks, respectively. These results indicate that E. adenophorum contains potent acaricidal ingredients against the hard tick H. longicornis.

Unusual 5/5 fused bicyclosesquiterpenoids from Eupatorium adenophorum.

The chemical analysis on the aerial sections of Eupatorium adenophorum Spreng. resulted in the identification of four unprecedented 5/5 fused bicyclosesquiterpenoids, eupatorid A (1), and its analogues named eupatorester A-C (2-4) using various chromatographic techniques. Their structures were unambiguously confirmed by detailed spectroscopic investigations (including 1D, 2D-NMR and HRMS), and single crystal X-ray diffraction. The anti-inflammatory activities, in vitro tumor growth inhibitory activities and antibacterial activities of these compounds were evaluated.

Bio-accumulation effects of heavy metals Pb, Zn and Cd on Procecidochares utilis parasitism to Eupatorium adenophorum at Suzu metal mines, Yunnan.

Procecidochares utilis is an obligatory parasitic insect to Eupatorium adenophorum. Both organisms have been spread to some metal mines areas. The objective of this study is to comprehend the trend of heavy metals transfer and the process of their bio-accumulation in the soil-E. adenophorum-P. utilis system and particularly their impact on the parasitic effect of P. utilis to E. adenophorum to reflect the impact of heavy metals on obligate parasitic insect and its host. Therefore, a detailed investigation was carried out at the Suzu Lead-Zinc Mine in Yunnan Province using the concentric circle's method. The results showed that the parasitic rate of P. utilis to a single plant and branch is positively correlated with distance. The metals content of the soil in E. adenophorum and P. utilis, decreased dramatically with an increase in distance away from the center of the mining area. From which is cleared that these metals could enter to E. adenophorum and P. utilis through the soil-E. adenophorum-P. utilis system which likely to affect its parasitic activities. In addition, the parasitic rate is impacted by per Zn content greatly, and the parasitic rate per plant is affected by Cd content enormously. This work could provide important basis of data for further understanding and clarifying the effects of bioaccumulation and heavy metals pollution on various aspects of the food chain. Simultaneously, it could clarify and simplify whether heavy metal contamination affects the parasitic behaviour of some obligatory parasitic insects.

Three new cadinane-type sesquiterpenes from Eupatorium adenophorum spreng.

Three new cadinane-type sesquiterpenes, eupatorinones A-C (1-3), along with seven known compounds (4-10), were obtained from the petroleum ether fraction of 95% ethanol extract of Eupatorium adenophorum Spreng. The structures of these new compounds were determined by NMR, MS, and ECD spectra analysis. The configuration of compound 3 was established by quantum chemical calculations of NMR chemical shifts and ECD spectra, that matched the experimental data. In addition, compounds 1, 3 and 5 increased the glucose uptake in L6 cells by 1.42, 1.21 and 1.60 times, respectively.[Formula: see text].

Removal of 2,4,6-trichlorophenol from water by Eupatorium adenophorum biochar-loaded nano-iron/nickel.

From the perspective of waste utilization, the invasive species, Eupatorium adenophorum was used to prepare biochar, which was then loaded with iron/nickel bimetals. Compared with pure biochar, the biochar-loaded nano-iron/nickel bimetals have a significant effect on the removal of 2,4,6-trichlorophenol (2,4,6-TCP) from water, and their degradation rate can be increased by 39.7%-71.6% under different conditions. Several factors can influence the removal of 2,4,6-TCP, including the load ratio, pH of the solution, concentration of 2,4,6-TCP, and coexisting ions in water (HCO3-, SO42-, NO3-). Based on the density functional model (DET), Ni can activate H2 (produced in the reaction between nano-Fe and H2O) to convert to H*, which can then substitute Cl. The activation energy is 109.5 kJ/mol, indicating the reaction is easy to take place.

[Soil microbial community characteristics in degraded karst tiankeng invaded by Eupatorium adenophorum.] / ç´«èŒŽæ³½å °å ¥ä¾µä¸‹å–€æ–¯ç‰¹é€€åŒ–å¤©å‘çš„åœŸå£¤å¾®ç”Ÿç‰©ç¾¤è½ç‰¹å¾.

After invading the grand negative landform karst degraded tiankeng, Eupatorium adenophorum will affect the plant diversity, community succession and natural ecological restoration of degraded tiankeng by changing soil microbial community. Taking the degraded tiankeng "Bajiaxiantang" as the research area, the soil around E. adenophorum roots in different habitats was selected to explore the microbial community characteristics of E. adenophorum roots using the Biolog-ECO micro-plate method. The results showed that the invasion degree of E. adenophorum was in order of pit slope shrub-grassland > pit bottom grassland > pit slope forest > pit head shrub-grassland. The characteristics of E. adenophorum varied among different habitats due to habitat heterogeneity. The change of microbial community metabolic activity, dominance index, richness index and evenness index of E. adenophorum roots was closely related with the invasion degree, with significant diffe-rence between inside and outside of the tiankeng. Amino acids and polymers were the main carbon sources utilized by soil microorganism. The α diversity of herbaceous plants in different habitats were correlated with that of soil microbial community. The diversity of soil microbial community around E. adenophorum roots increased significantly with the increases of invasion level, which could form a microenvironment conducive to invasion and colonization. In the future, the degraded tiankeng would eventually form the climax community of underground forests with community succession and natural ecological restoration, which might result in the disappearance of E. adenophorum in the degraded tiankeng.

Essential Oil Derived From Eupatorium adenophorum Spreng. Mediates Anticancer Effect by Inhibiting STAT3 and AKT Activation to Induce Apoptosis in Hepatocellular Carcinoma.

Eupatorium adenophorum Spreng. (EA) is a well-known noxious invasive species. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that the essential oil derived from EA (EAEO) is mainly composed of sesquiterpenes. However, the pharmacological value of EAEO in hepatocellular carcinoma (HCC) remains largely unexplored. Herein, we investigated the anti-HCC activities of EAEO, and explored the potential mechanisms of EAEO-induced apoptosis. An MTT assay showed that EAEO inhibited HCC cell proliferation with little toxicity on normal liver cells. Wound healing and FACS assays revealed that EAEO suppressed HCC cell migration and arrested cell cycle, respectively. Moreover, EAEO promoted in vitro HCC cell apoptosis, and EAEO treatment inhibited HepG2 xenografts growth and enhanced apoptotic nucleus of xenografts in HepG2-bearing nude mice. Mechanistically, EAEO significantly decreased the ratio of Bcl-2/Bax and resulted in the activation of caspase-9 and -3. EAEO also reduced the expression of Grp78, which in turn relieved the inhibition of caspase-12 and -7. Meanwhile, EAEO suppressed the phosphorylation of STAT3 and AKT, indicative of its anti-HCC potential. In summary, we determined that EAEO treatment promoted HCC apoptosis via activation of the apoptotic signaling pathway in mitochondria and endoplasmic reticulum, as well as repressed the activity of STAT3 and AKT in HCC cells.

[Exogenous NO application effectively alleviates the allelochemical stress on cucumber root border cells caused by Eupatorium adenophorum extracts.] / 外源NOç¼“è§£ç´«èŒŽæ³½å °æå–ç‰©å¯¹é»„ç“œæ ¹è¾¹ç¼˜ç»†èƒžçš„åŒ–æ„Ÿèƒè¿«.

Using suspension cultures of cucumber (Cucumis sativus) cultivar Jinyou 35, we investigated the effects of allelochemical stresses induced by Eupatorium adenophorum extracts on root border cells (RBC), and the role of exogenous NO application in alleviation of the damage of root tips exposed to E. adenophorum extracts. The results showed that, 1000 mg·L-1 E. adenophorum extracts had significant damage to the cucumber root tip, resulting in severe tissue damage, exfoliated surface cells and irregular arrangement of inner cells, while those damages could be effectively alleviated by spraying exogenous NO. Compared with the control, E. adenophorum extracts (ZL) markedly reduced RBC numbers and survival rates by 54.5% and 97.2%, respectively, the RBC apoptosis rates were 12.3 times higher, the thicknesses of RBC adhesive layers were increased by 31.4%, and the root cap PME activities were markedly increased. Compared with the ZL treatment, exogenous NO application (ZN) significantly increased RBC numbers and survival rates by 72.4% and 146.0%, respectively, reduced the corresponding RBC apoptosis rates and the thicknesses of RBC adhesive layers by 30.7% and 15.0%, respectively, and inhibited the PME activities by 14.3% upon treatment for 72 hours. Our findings revealed that E. adenophorum extracts showed toxic effects on the cucumber RBC, resulting in cell apoptosis, abolishment of the RBC protection on root tips, and the destruction of root tip structure. Exogenous NO application, to some extent, could prevent the root tip and RBC from cell damage caused by E. adenophorum extracts.

Hepatotoxicity of Eupatorium adenophorum extracts and the identification of major hepatotoxic components.

Eupatorium adenophorum can induce liver toxicity in animals. For the safe utilisation of the weed, the hepatotoxic components need to be discovered. In this study, in vitro hepatotoxicity of different extracts from E. adenophorum were determined on human hepatocyte cell line L02 and hepatocellular carcinoma cell line HepG2. The results showed that water extracts of E. adenophorum exhibited no hepatotoxicity in vitro while high concentrations of the organic solvent extracts had obvious hepatotoxicity. Sesquiterpenes may contribute to the toxicity based on the comparison of composition analysis. Three cadinene sesquiterpenes were purified and identified as 9-oxo-10,11-dehydroageraphorone, 10Hα-9-oxo-ageraphorone and 10Hß-9-oxo-ageraphorone. In vitro hepatotoxic effects of these components were investigated, the IC50 of the three compounds were 122.53, 87.52, and 108.80 µM in L02 cells and 151.92, 104.48, and 138.08 µM in HepG2 cells by Cell Counting Kit-8 (CCK-8) assay. The three components were confirmed to be, at least partial, hepatotoxic components.

Enrichment and separation of chlorogenic acid from the extract of Eupatorium adenophorum Spreng by macroporous resin.

A simple and efficient chromatographic method for separation of chlorogenic acid from Eupatorium adenophorum Spreng extract was developed. The adsorption properties of nine macroporous resins were evaluated. NKA-II resin showed much better adsorption/desorption properties. The adsorption of chlorogenic acid on NKA-II resin at 25°C was well fitted to Langmuir isotherm model and pseudo-second-order kinetic model. The dynamic adsorption and desorption experiments were carried out on columns packed with NKA-II resin to optimize the separation process. The content of chlorogenic acid in the product increased to 22.17%, with a recovery yield of 82.41%.

A new cadinane-type sesquiterpenoid from Eupatorium adenophorum Spreng / 药学学报

Four cadinane-type sesquiterpenes were obtained from the petroleum ether of 95% ethanol extract of Eupatorium adenophorum Spreng by using an HP-20 macroporous resin column, silica gel, and semi-preparative HPLC. Their structures were determined by physical, chemical and spectroscopic methods and identified as eupatorinol (1), (+)-(5R,7S,9R,10S)-2-oxocadinan-3,6(11)-dien-12,7-olide (2), (1S,4R)-7-hydroxycalamenen-3-one(3) and (-)-(5R,6R,7S,9R,10S)-cadinan-3-ene-6,7-diol (4). Among them, compound 1 is a new cadinane-type sesquiterpene, and compound 3 was isolated from this genus for the first time. In bioassay, none of these compounds displayed obvious cytotoxicity.

Chemical constituents from ethyl acetate extract of Eupatorium adenophorum / 中草药

Objective: To study the chemical constituents of the Eupatorium adenophorum. Methods: The chemical constituents were isolated and purified by silica gel chromatography repeatedly from the ethyl acetate extract of E. adenophorum, and their structures were identified by spectral analysis and chemical methods. Results: Fourteen compounds were isolated from E. adenophorum and identified as p-hydroxybenzoic acid ethyl ester (1), (1R,4R)-8aα-hydroxy-1-isopropyl-4,7-dimethyl- 1,2,3,4,6,8a-hexahydro-naphthalene-2,6-dione (2), daedalin A (3), 10-oxo-7-hydroxy-nordehydrotremetone (4), caffeoyl acetate (5), syringic acid (6), 3-hydroxy-4-(1-oxo-ethane)-benzoic acid (7), ferulic acid (8), p-hydroxyphenylethyl alcohol (9), protocatechuic acid ethyl ester (10), 4-hydroxy-3-isopropyl benzoic acid (11), balanophonin (12), indole-3-carboxylic acid (13), and 6-methoxy kaempferol (14). Conclusion: Compound 11 is obtained from natural products for the first time, compounds 1, 3, 6, 7, 9, 10, 12-14 are obtained from this genus for the first time, and compound 5 is obtained from this plant for the first time.