Leaf phenotypic variation and its response to environmental factors in natural populations of Eucommia ulmoides.

BACKGROUND: Eucommia ulmoides leaves have high medicinal and economic value as a dual-purpose substance for medicine and food. Employing leaves from 13 natural populations of Eucommia ulmoides as research objects, this study reveals the variation patterns of intra-specific and inter-specific trait variation and explores the response of leaf characteristics to geographical and climatic changes, aiming to provide a scientific basis for the efficient utilization of leaf resources and the breeding of superior varieties. RESULTS: Descriptive statistical analysis and nested analysis of variance showed significant differences in 11 leaf traits of Eucommia ulmoides inter-populations and intra-populations, with an average coefficient of variation of 17.45%. The coefficient of variation for average leaf phenotypic traits is 20.77%, and the leaf phenotypic variation is mainly from the variation intra-populations. Principal component analysis reveals that the cumulative contribution rate of the top three principal components which mainly contributed to the phenotypic variation of Eucommia ulmoides leaves reached 74.98%, which could be sorted into size traits (34.57%), color traits (25.82%) and shape traits (14.58%). In addition, correlation analysis expresses there is a specific co-variation pattern among leaf traits, with a strong connection between shape, size, and color traits. Geographic and climatic distances are significantly correlated, and mantel test and correlation analysis indicate that leaf traits of Eucommia ulmoides are mainly influenced by altitude. With the increase of altitude, the leaves become smaller. Partial correlation analysis shows that after controlling climate factors, the correlation between some characters and geographical factors disappears significantly. Temperature and precipitation have a great influence on the variation of leaf phenotypic traits, and the larger the leaves are in areas with high temperature and heavy rainfall. CONCLUSIONS: These findings contribute to a further understanding of the leaf morphological characteristics of Eucommia ulmoides and the extent to which the environment influences leaf trait variation. They can provide a scientific basis for the protection and application of Eucommia ulmoides leaf resources in the future.

Extraction technology, components analysis and anti-inflammatory activity in vitro of total flavonoids extract from Artemisia anomala S. Moore.

Artemisia anomala S. Moore exerts many pharmacological activities, including the removing of the blood stasis, relieving of the fever and analgesia, reducing the swelling and dampness. In this study, the extraction technology, chemical compositions and anti-inflammatory effect in vitro and mechanism of total flavonoids extract from Artemisia anomala S. Moore were studied. The optimal yield rate of total flavonoids extract was optimized by single factor experiments and response surface method, and the chemical constituents were analyzed by UPLC-QTOF-MS method; and the anti-inflammatory activity of the extract was evaluated with lipopolysaccharide induced RAW 264.7 cells. The highest extraction rate was 2.02% under these conditions of the concentration of ethanol 50%, the ultrasonic extraction time 30 min, and the ratio of solvent volume to material weight 20:1 (ml/g). In addition, the main components of total flavonoid extract were preliminarily identified and deduced based on mass spectrometry information and relevant literatures, and its stronger anti-inflammatory activity was demonstrated by reducing the phagocytosis, the content of nitric oxide and the level of related cytokines (tumor necrosis factor-α, interleukin-10, interleukin-6). Furthermore, it was further revealed that the anti-inflammatory effect of the extract was closely connected with the activation of TLR4-MyD88-NF-κB signalling pathway. This study indicated that the total flavonoids extract from Artemisia anomala S. Moore may be a better candidate anti-inflammatory natural medicine.

Polyethylene glycol modified graphene oxide-silver nanoparticles nanocomposite as a novel antibacterial material with high stability and activity.

Inorganic antibacterial nanomaterials play an increasingly important role in addressing the growing threat of drug-resistant bacteria. Graphene oxide-silver nanoparticles composite (GO-AgNPs), as a kind of inorganic nanomaterials, have excellent antibacterial properties, showing promising potential in biomedical field. However, GO-AgNPs are terribly prone to sedimentation due to aggregation in physiological solutions, along with its non-environmental issues during the synthesis process, seriously limits the antibacterial application of GO-AgNPs in the biomedical field. To solve this problem, herein, polyethylene glycol-graphene oxide-silver nanoparticles composite (GO-AgNPs-PEG) were prepared by modifying GO-AgNPs with polyethylene glycol to enhance their dispersion stability in physiological solutions. In addition, GO-AgNPs-PEG were prepared with using the natural product gallic acid as a reductant and stabilizer, exhibiting the characteristic of environmentally friendly. Meanwhile, the dispersion stability and antibacterial activity of GO-AgNPs-PEG were characterized by various technical methods, it was found that GO-AgNPs-PEG can be stably dispersed in a variety of physiological solutions (e.g., physiological saline, phosphate buffer solution, Luria-Bertani medium, Murashige and Skoog medium) for more than one week. Moreover, the antibacterial properties of GO-AgNPs-PEG in physiological solutions were significantly better than those of GO-AgNPs. Furthermore, it was discovered that the antibacterial mechanism of GO-AgNPs-PEG was probably associated to destroying the integrity of bacterial cell walls and membranes. The findings in this work can provide new ideas and references for the development of new inorganic antibacterial nanomaterials with stable dispersion in physiological solutions.

Analysis on the thermal decomposition kinetics and storage period of biomass-based lycorine galanthamine.

As global ageing deepens and galanthamine is the preferred clinical drug for the treatment of mild to moderate Alzheimer's disease, it will be valuable to examine the behaviour and mechanism of galanthamine's thermal decomposition for its quality control, formulation process, evaluation of thermal stability, and expiry date in production. In order to study the pyrolysis of galanthamine hydrobromide with nitrogen as the carrier gas, a thermogravimetric-differential thermogravimetric technique (TG-DTG) was applied at a temperature rise rate of 10 K min-1 and a volume flow rate of 35 mL min-1. The apparent activation energy E a and the prefactor A (E a = 224.45 kJ mol-1 and lnA = 47.40) of the thermal decomposition reaction of galanthamine hydrobromide were calculated according to the multiple heating rate method (Kissinger and Ozawa) and the single heating rate method (Coats-Redfern and Achar), and the most probable mechanism function was derived, and then the storage period was inferred from E a and E. A three-dimensional diffusion mechanism was suggested to control the thermal decomposition of galanthamine hydrobromide in accordance with the Jander equation, random nucleation and subsequent growth control, corresponding to the Mample one-way rule and the Avrami-Erofeev equation. As a result, the thermal decomposition temperature of galanthamine hydrobromide gradually increased with the rate of temperature rise. From Gaussian simulations and thermogravimetric data, galanthamine hydrobromide decomposed at the first stage (518.25-560.75 K) to release H2O, at the second stage (563.25-650.75 K) to generate CO, CO2, NH3 and other gases, and finally at the third stage (653.25-843.25 K) to release CO2. After 843.25 K, the residual molecular skeleton is cleaved to release CO2 and H2O. According to the E a and A presenting in the first stage of thermal decomposition, it is assumed that the storage life of galanthamine hydrobromide at room temperature 298.15 K is 4-5 years.

Effects of Luteolin on Biofilm of Trueperella pyogenes and Its Therapeutic Effect on Rat Endometritis.

Trueperella pyogenes is an opportunistic pathogen that causes suppurative infections in animals. The development of new anti-biofilm drugs will improve the current treatment status for controlling T. pyogenes infections in the animal husbandry industry. Luteolin is a naturally derived flavonoid compound with antibacterial properties. In this study, the effects and the mechanism of luteolin on T. pyogenes biofilm were analyzed and explored. The MBIC and MBEC of luteolin on T. pyogenes were 156 µg/mL and 312 µg/mL, respectively. The anti-biofilm effects of luteolin were also observed by a confocal laser microscope and scanning electron microscope. The results indicated that 312 µg/mL of luteolin could disperse large pieces of biofilm into small clusters after 8 h of treatment. According to the real-time quantitative PCR detection results, luteolin could significantly inhibit the relative expression of the biofilm-associated genes luxS, plo, rbsB and lsrB. In addition, the in vivo anti-biofilm activity of luteolin against T. pyogenes was studied using a rat endometritis model established by glacial acetic acid stimulation and T. pyogenes intrauterine infusion. Our study showed that luteolin could significantly reduce the symptoms of rat endometritis. These data may provide new opinions on the clinical treatment of luteolin and other flavonoid compounds on T. pyogenes biofilm-associated infections.

Molecular Basis for Luteolin as a Natural TatD DNase Inhibitor in Trueperella pyogenes.

TatD960 and TatD825 are DNases that contribute to biofilm formation and virulence in Trueperella pyogenes (T. pyogenes). Luteolin is a natural flavonoid commonly found in plants that exhibits antimicrobial capacity. Our study aims to investigate the effects of luteolin on TatD DNases as a natural inhibitor. In this research, the expression of tatD genes and TatD proteins in T. pyogenes treated with luteolin was detected, and then the effect of luteolin on the hydrolysis of DNA by TatD DNases was analyzed using agarose gel electrophoresis. Moreover, the interactions between luteolin and TatD DNases were tested using surface plasmon resonance (SPR) assays and molecular docking analysis. After 1/2 MIC luteolin treatment, the transcription of tatD genes and expression of TatD proteins appeared to be reduced in 80-90% of T. pyogenes (n = 20). The gel assay revealed that luteolin can inhibit the activity of TatD DNases. The SPR assay showed that the KD values of luteolin to TatD960 and TatD825 were 6.268 × 10-6 M and 5.654 × 10-6 M, respectively. We found through molecular docking that hydrogen bonding is predominant in the interaction of luteolin and TatD DNases. Our data indicate that luteolin inhibited the ability of TatD DNases by decreasing their binding to DNA. The current study provides an insight into the development of luteolin as a DNase inhibitor in preventing biofilm formation and virulence in T. pyogenes.

In Situ Green Synthesis of Graphene Oxide-Silver Nanoparticles Composite with Using Gallic Acid.

The adoption of plant-derived natural products to synthesize metal nanoparticles and their complexes has the advantages of mild reaction conditions, environmental protection, sustainability and simple operation compared with traditional physical or chemical synthesis methods. Herein, silver nanoparticles (AgNPs) were in situ synthesized on the surface of graphene oxide (GO) by a "one-pot reaction" to prepare graphene oxide-silver nanoparticles composite (GO-AgNPs) based on using AgNO3 as the precursor of AgNPs and gallic acid (GA) as the reducing agent and stabilizer. The size and morphology of GO-AgNPs were characterized by ultraviolet-visible spectrophotometer (Uv-vis), Fourier transform infrared spectroscopy (FT-IR), transmission electron microscope (TEM), X-ray diffractometer (XRD) and dynamic light scattering (DLS). The effects of pH, temperature, time and material ratio on the synthesis of GO-AgNPs were investigated experimentally. The results showed that ideal GO-AgNPs could be prepared under the conditions of pH = 9, 45°C, 2 h and the 2:1 of molar ratio of AgNO3 to GA. The AgNPs within GO-AgNPs are highly crystalline spherical particles with moderate density on the surface of GO, and the size of AgNPs is relatively uniform and determined to be about 8.19 ± 4.21 nm. The research results will provide new ideas and references for the green synthesis of metal nanoparticles and their complexes using plant-derived natural products as the reducing agent and stabilizer.

The protective effect of rutin against lipopolysaccharide induced acute lung injury in mice based on the pharmacokinetic and pharmacodynamic combination model.

Rutin is a flavonoid compound with many pharmacological activities, including antioxidation, anti-inflammation and cardiovascular and cerebrovascular protection. However, there are great limitations in clinical application in view of its poor solubility and slow absorption in vivo. In this study, a pharmacokinetic and pharmacodynomic model was adopted to study the correlation of the pharmacokinetics and pharmacodynomics of rutin in lipopolysaccharide-induced acute lung injury mice. Rutin was intragastrically administered continuously for 5 days at a dose of 200 mg/kg/d, and pharmacokinetic and pharmacodynamic indicators were measured every day after administration, including the blood concentration of rutin, the W/D ratio of lungs, the nitric oxide content and the expression levels of TLR4, TRAF6, IκB and P-IκB proteins. The results indicated that rutin can exert an anti-inflammatory protective effect by improving lung tissue injury, significantly decreasing the synthesis of the inflammatory mediator nitric oxide, and inhibiting the protein expression levels of TLR4, TRAF6 and P-IκB. The absorption of rutin conformed to a one-compartment model with the pharmacokinetic parameters as follows: t1/2α= 9.76 h, t1/2ß= 19.44 h, Tmax= 24.00 h, Cmax= 22.65 µg/ml and AUC(0-t)= 518.58 µg/ml·h. A PK-PD combination model was established to fit the optimal administration time of rutin with a one-compartment-Sigmod Emax model connected to the effect site. Meanwhile,the PK-PD combination model was a better approach for evaluating the relationships between the five pharmacodynamic indicators and the pharmacokinetic characteristics of rutin. The correlation between the pharmacokinetics and pharmacodynamics of rutin was quantitatively analysed to provide a theoretical basis for the research and development of new anti-inflammatory drugs in clinical practice.

TatD DNases Contribute to Biofilm Formation and Virulence in Trueperella pyogenes.

TatD DNases are conserved proteins in a variety of organisms and are considered potential virulence factors in Plasmodium falciparum and Streptococcus pneumoniae. However, the function of TatD DNases has not yet been determined in Trueperella pyogenes, which causes various infections in animals and leads to economic losses. In this study, we describe the roles of TatD DNases in T. pyogenes (TpTatDs). A bioinformatics analysis was performed to investigate the sequence characteristics of TpTatDs, and then the ability of recombinant TatD proteins to hydrolyze DNA was determined in the presence of divalent cations. Moreover, we constructed tatD-deficient mutants. The biofilms formed by the wild-type and mutant strains were observed under a microscope. The mortality and bacterial load in the spleen of mice infected with the wild-type strain and tatD-deficient mutants were determined to obtain insights into the role of TatDs in the virulence of T. pyogenes. Two TatD DNases were identified in T. pyogenes. They were Mg2+-dependent DNases and exhibited DNA endonuclease activity. Compared with those formed by the parental strain, biofilms formed by mutants showed a significantly reduced thickness and biomass. Moreover, mutants produced a lower bacterial load in the spleen of mice and compromised virulence. Our data indicated that TatD DNases in T. pyogenes are involved in biofilm formation and required for virulence during infections.

Optimization of ultrasound extraction of Tribulus terrestris L. leaves saponins and their HPLC-DAD-ESI-MSn profiling, anti-inflammatory activity and mechanism in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Tribulus terrestris L., as an annual herb plant from Zygophyllaceae, exhibits many biological activities, and its main chemical constituents are saponins. However, the extraction process, chemical compositions, anti-inflammatory effect and mechanism of total saponins from Tribulus terrestris L. leaves are still unclear. AIM OF THE STUDY: The present study extensively evaluated the extraction process, major components, anti-inflammatory action and mechanism of Tribulus terrestris L. leaves saponins. MATERIALS AND METHODS: The ultrasonic extraction and response surface methods were adopted for optimization of extraction technology of total saponins from Tribulus terrestris L. leaves, and its compositions were detected with LC-MSn method. The anti-inflammatory activity of total saponins was studied by lipopolysaccharide induced RAW 264.7 cells and acute lung injury mice models. RESULTS: The ultrasonic extraction parameters of saponins fraction, including ethanol concentration 30%, extraction time 55 min, ratio of solvent to material 35:1 ml/g and extraction temperature 46 °C, were screened by response surface method with the extracting rate 5.49%, and thirty compositions were detected with LC-MSn method. Moreover, saponins fraction can play a stronger anti-inflammatory effect by reducing the phagocytic activity and pulmonary edema, and protection of morphology of RAW 264.7 cells and lung tissues, and decreasing the content of NO and TNF-α. Moreover, it was revealed that total saponins extract can exert the anti-inflammatory action by the inhibition of the activation of the TLR4-TRAF6-NF-κB signalling pathway. CONCLUSION: These studies imply that Tribulus terrestris L. leaves saponins may be an important anti-inflammatory drug in clinic.

Rutin prevents inflammation induced by lipopolysaccharide in RAW 264.7 cells via conquering the TLR4-MyD88-TRAF6-NF-κB signalling pathway.

OBJECTIVES: Inflammation widely exists in many diseases and poses a great threat to human and animal health. Rutin, quercetin-3-rhamnosyl glucoside, has a variety of pharmacological effects, including anti-oxidant, anti-inflammatory, antibacterial, anticancer and radioresistance effects. The current study focused on evaluation of its anti-inflammatory activity and described the mechanism of rutin in lipopolysaccharide-induced RAW 264.7 cells. METHODS: The related gene and protein expression levels were investigated by quantification real-time PCR and western blotting, respectively. KEY FINDINGS: This study revealed that rutin can decrease inducible nitric oxide synthase (iNOS) gene and protein expression levels, effectively increase IκB gene expression, reduce toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), tumour necrosis factor receptor-associated factor 6 (TRAF6) and p65 gene expression and inhibit the phosphorylation of IκB and p65 and the proteins expression of TLR4, MyD88 and TRAF6. CONCLUSIONS: These results suggest that rutin might exert anti-inflammatory effect on LPS-stimulated RAW 264.7 cells and will be potentially useful as an adjuvant treatment for inflammatory diseases.

Novel Protein Tyrosine Phosphatase 1B Inhibitor-Geranylated Flavonoid from Mulberry Leaves Ameliorates Insulin Resistance.

Mulberry leaf is a common vegetable with a variety of beneficial effects, such as hypoglycemic activity. However, the underlying mechanism of its hypoglycemic effect have not been fully revealed. In this study, two flavonoid derivatives were isolated from mulberry leaves, a new geranylated flavonoid compound (1) and its structural analogue (2). The structures of compounds 1 and 2 were elucidated using spectroscopic analysis. To study the potential hypoglycemic properties of these compounds, their regulatory effects on protein tyrosine phosphatase 1B (PTP1B) were investigated. In comparison to oleanolic acid, compounds 1 and 2 showed significant inhibitory activities (IC50 = 4.53 ± 0.31 and 10.53 ± 1.76 µM) against PTP1B, the positive control (IC50 = 7.94 ± 0.76 µM). Molecular docking predicted the binding sites of compound 1 to PTP1B. In insulin-resistance HepG2 cell, compound 1 promoted glucose consumption in a dose-dependent manner. Furthermore, western blot and polymerase chain reaction analyses indicated that compound 1 might regulate glucose consumption through the PTP1B/IRS/PI3K/AKT pathway. In conclusion, geranylated flavonoids in mulberry leaves inhibite PTP1B and increase the glucose consumption in insulin-resistant cells. These findings provide an important basis for the use of mulberry leaf flavonoids as a dietary supplement to regulate glucose metabolism.

The Antibacterial Activity and Mechanism of Action of Luteolin Against Trueperella pyogenes.

PURPOSE: This research aimed to investigate the antibacterial activity and potential mechanism of luteolin against T. pyogenes. MATERIALS AND METHODS: The broth microdilution method was used to determine the minimum inhibitory concentrations (MICs) of luteolin against various T. pyogenes strains. The potential mechanism of action of luteolin was elucidated through testing and analysing the luteolin-induced alterations of T. pyogenes in several aspects, including cell wall, cell membrane, protein expression, nucleic acid content, topoisomerase activity and energy metabolism. RESULTS: The MIC values of luteolin against various T. pyogenes isolates and ATCC19411 were 78 µg/mL. The increased cell membrane permeability, destruction of cell wall integrity and TEM images after exposure to luteolin showed that the cell wall and membrane were damaged. The content of total protein and nucleic acid in T. pyogenes decreased significantly after treatment with luteolin (1/2 MIC) for 12, 24, and 36 h. Moreover, a hypochromic effect was observed in the absorption spectrum of luteolin when deoxyribonucleic acid (DNA) was added. In addition, after treatment with luteolin, a decrease in nicked or relaxed DNA content, which was catalysed by T. pyogenes-isolated DNA topoisomerase, was observed. In addition, the adenosine triphosphate (ATP) content in cells and the activity of succinate dehydrogenase (SDH) both decreased when T. pyogenes was exposed to different concentrations (1/4 MIC, 1/2 MIC, 1 MIC, 2 MIC) of luteolin for 1 h. CONCLUSION: Luteolin showed distinct antibacterial activity against T. pyogenes by multiple actions, which mainly include destroying the integrity of the cell wall and cell membrane, influencing the expression of proteins, inhibiting nucleic acid synthesis, and interfering with energy metabolism.

In vitro Antibiotic Susceptibility, Virulence Genes Distribution and Biofilm Production of Staphylococcus aureus Isolates from Bovine Mastitis in the Liaoning Province of China.

PURPOSE: The aim of this study was to identify the subtype, characterize the antimicrobial resistance, determine the virulence gene distribution, and analyze the biofilm production of Staphylococcus aureus isolates from bovine mastitis milk samples in the Liaoning Province of China. MATERIALS AND METHODS: In total, 56 Staph. aureus isolates were collected and identified in this study; the isolates were divided into different spa types based on the sequence of the polymorphic X region of the spa gene. Additionally, antimicrobial susceptibility was investigated using the broth microdilution method, and 18 virulence genes were detected using PCR. Biofilm formation was measured by spectrophotometry with crystal violet staining and observed using confocal laser scanning microscopy. RESULTS: There were 12.12% (56/462) milk samples that were positive for Staph. aureus. These isolates were nonsusceptible to sulfamethoxazole (100%), penicillin (76.9%), daptomycin (76.79%), clindamycin (69.64%), and oxacillin (60.71%); however, the majority of the isolates (80.4%) were susceptible to amoxicillin/clavulanate. The predominant virulence genes encoded the cytotoxins, hla (94.64%) and hlb (89.29%), and the adhesion factors clfA (89.29%), clfB (89.29%), and fnbB (80.36%). Comparatively, virulence genes related to other adhesion factors such as cna (8.93%) and enterotoxins, such as seg (26.79%), sea (16.07%), seb (7.14%), and sec (7.14%) were detected at relatively lower rates. The following eight spa types were identified: t267 (35.84%), t730 (22.64%), t518 (15.09%), t1190 (11.32%), t1456 (9.43%), t224 (1.88%), t9129 (1.88%), and t177 (1.88%). The highest biofilm production was observed for t267. Staph. aureus exhibited various patterns of biofilm formation, with the biofilm often being associated with a tower-shaped structure or a thicker biofilm. CONCLUSION: Our results indicated that Staph. aureus isolates from dairy cows with mastitis in the Liaoning Province of China were non-susceptible to sulfamethoxazole, penicillin, daptomycin, oxacillin, and clindamycin. Additionally, the most prevalent subtype was t267, which displayed resistance to multiple antimicrobial agents and harbored several virulence genes, including clfA, clfB, fnbB, hla, and hlb.

Response surface optimization and antioxidant activity of total proanthocyanidins fraction from Abutilon theophrasti Medic. leaves.

The extraction procedure and antioxidant activity were investigated for total proanthocyanidins extracts from Abutilon theophrasti Medic. leaves collected in August, September and October. The maximum extraction yield was achieved with 90% ethanol, 80°C of heating reflux temperature, 149.94 min of extraction time and 60(ml/g) of the ratio of solvent and material, which were optimized by Box-Behnken Design of response surface method. Spectrophotometric study displayed that total proanthocyanidins content was (0.44±0.02)% (0.52±0.01)% and (0.59±0.01)% for August, September and October samples, respectively. The proanthocyanidins extracts exhibited much stronger antioxidant activity to scavenge ABTS and DPPH free radicals, and reduce ferric power than the control synthetic antioxidant BHT. The present findings suggest that the proanthocyanidins extract from Abutilon theophrasti Medic. leaves was a very interesting candidate for the research and development of natural and healthy antioxidant for the pharmaceutical and food industries.

Extraction technology, component analysis, antioxidant, antibacterial, analgesic and anti-inflammatory activities of flavonoids fraction from Tribulus terrestris L. leaves.

Research on the extraction technology, component analysis, and antioxidant, antibacterial, analgesic and anti-inflammatory activities of flavonoids fraction from Tribulus terrestris L. leaves was carried out. The extraction process was optimized by response surface method, and the maximum yield 0.27% for flavonoids fraction was achieved with concentration of ethanol solution 25.87%, ratio of solvent to material 30:1 ml/g, and ultrasonic extraction time 27.93 min. Moreover, 14 compositions were identified separately from flavonoids fraction by HPLC-DAD-ESI-MS2. In addition, flavonoids fraction exhibited a better antioxidant activity demonstrated by assays of ABTS, DPPH radical scavenging activity and ferric reducing antioxidant power activity. Furthermore, the antibacterial, analgesic and anti-inflammatory activities of flavonoids fraction were also proved to be stronger. Therefore, the present findings suggest that flavonoids fraction from Tribulus terrestris L. leaves can be a very interesting candidate for the research and development of natural and healthy herbal medicine for the pharmaceutical and food industries.

The regulatory effect of flavonoids extracted from Abutilon theophrasti leaves on gene expression in LPS-induced ALI mice via the NF-κB and MAPK signaling pathways.

Context: ALI is a common disease characterized by acute pulmonary inflammatory disorder. Abutilon theophrasti Medik. (Malvaceae), as a Chinese traditional medicine, is used for the treatment of inflammation. Its main constituents are flavonoid compounds. Objective: This study investigates the regulatory effect of a TFE from Abutilon theophrasti leaves on gene expression in LPS-induced ALI mice via the NF-κB and MAPK signaling pathways. Materials and methods: Kunming mice were intragastrically administered TFE (0.25, 0.5, 1.0 g/kg) for 5 days, and then ALI was induced via intranasal administration 40 µg of LPS in 10 µL PBS after intragastric administration on the 5th day, and PBS and DEX (2 mg/kg) were negative and positive control groups, respectively. Results: The relative expression of iNOS gene was 0.707, 0.507 and 0.483 for 0.25, 0.5 and 1.0 g/kg TFE, and COX-2 gene expression was also reduced after treatment by three concentrations of TFE with 0.768, 0.545, and 0.478. The mRNA expression levels of p65 were 0.61, 0.43 and 0.27 for 0.25, 0.5 and 1.0 g/kg TFE and IκB levels were increased in a dose-dependent manner with 3.99, 13.69 and 34.36. 0.5 and 1.0 g/kg TFE inhibited the expression of ERK1/2 with 0.59 and 0.38, p38MAPK with 0.62 and 0.54, and JNK with 0.37 and 0.29, and JNK mRNA expression was 0.60 for 0.25 g/kg TFE. Discussion and conclusion: These results indicate that the regulatory mechanisms of TFE on gene expression in LPS-induced ALI mice include inhibition of the NF-κB and MAPK signaling pathways.

Luteolin Showed a Resistance Elimination Effect on Gentamicin by Decreasing MATE mRNA Expression in Trueperella pyogenes.

Trueperella pyogenes is a common inhabitant of mucosal surfaces in animals and causes a variety of infections, including endometritis, mastitis, and liver abscessation, in dairy cows. Many antimicrobial agents are used for treatment of infections caused by T. pyogenes; however, antibiotic resistance has recently become a serious problem. The objective of this study was to characterize the effect of the efflux pump-encoding multidrug and toxic compound extrusion (MATE) gene on antibiotic resistance in T. pyogenes isolates from cows with signs of endometritis. As a compound from plants, luteolin showed antimicrobial activities in Escherichia coli and Staphylococcus aureus; therefore, we also investigated whether luteolin can eliminate antibiotic resistance. We constructed a MATE deletion mutant in BM-H06-3 to identify the function of MATE in antibiotic resistance. MATE mRNA expression was measured to identify the mechanism of luteolin in gentamicin resistance elimination effect in T. pyogenes. The T. pyogenes isolate BM-H06-3 became susceptible to gentamicin, amikacin, streptomycin, erythromycin, and roxithromycin after MATE deletion. No synergistic effect between luteolin and gentamicin was observed in eight isolates, which were randomly selected from 34 T. pyogenes isolates, but the isolates became susceptible to gentamicin after luteolin treatment at a subinhibitory concentration (1/4 minimum inhibitory concentration [MIC]) for 36 hr. Furthermore, luteolin can decrease MATE mRNA expression after luteolin treatment at a subinhibitory concentration (1/4 MIC). We found that the MATE gene was involved in antibiotic resistance and that luteolin induces a resistance elimination effect in T. pyogenes. Therefore, luteolin may be a potential agent to inhibit efflux pumps in multidrug-resistant T. pyogenes.

Extraction technology, component analysis, and in vitro antioxidant and antibacterial activities of total flavonoids and fatty acids from Tribulus terrestris L. fruits.

The current study focused on the extraction technology, components analysis, in vitro antioxidant and antibacterial activities of total flavonoids and fatty acids from Tribulus terrestris L. fruits. The extraction process of total flavonoids and fatty acids was optimized by the response surface method, and the compositions were identified from the two extracts by HPLC-DAD-ESI-MS- and GC-MS, respectively. In addition, the antioxidant and antibacterial activities were evaluated by assay of ABTS, DPPH radical scavenging activity, ferric reducing antioxidant power and minimal inhibitory concentration. The yields of total flavonoids and fatty acids were 0.46 and 9.76% under the optimized conditions. Moreover, nine and eight compositions were identified from the two extracts based on the related references, respectively. In addition, total flavonoids and fatty acids extracts both exhibited certain antioxidant and antibacterial activities. The present findings suggest that total flavonoids extracted from T. terrestris L. fruits comprised a more interesting candidate than fatty acids for the research and development of natural and healthy antioxidants and antibacterial agents for the pharmaceutical and food industries.

The protective effect of the flavonoid fraction of Abutilon theophrasti Medic. leaves on LPS-induced acute lung injury in mice via the NF-κB and MAPK signalling pathways.

Accompanied by the damages of epithelial and capillary endothelial cell, acute lung injury is diagnosed with the typical pathological symptoms in clinic, including diffusing of pulmonary interstitial, alveolar oedema and hypoxic respiratory insufficiency. Current study focused on the investigation the anti-inflammatory action and mechanisms of total flavonoids extract (TFE) from Abutilon theophrasti Medic. leaves on ALI mice induced by LPSs. Mice were administrated intragastrically with TFE at the concentrations of 0.25, 0.5, or 1.0 g/kg for 5 days, and on last day, nasal administration of LPSs for 6 h after 30 min for intragastric administration of TFE. Pretreatment with TFE not only reduced oxidative damage but also alleviated lung edema in ALI mice. Increased secretion of pro-inflammatory cytokines TNF-α, IL-1ß and IL-6, caused by LPSs was reversed by TFE; on the contrary, the anti-inflammatory cytokine IL-10 was upregulated. The proteins expressions of pro-inflammatory mediators iNOS and COX-2 induced by LPSs, were down-regulated by TFE. Moreover, the activation of NF-κB and MAPK signalling pathways was inhibited by TFE in LPSs induced ALI mice. The results revealed that the anti-inflammatory mechanisms of TFE were via inhibition of NF-κB and MAPK activation. Combined, the results suggested that TFE might exert in vivo antioxidant and anti-inflammatory functions in LPSs stimulated mice, and will be potential in adjuvant treatment in oxidative stress and inflammation diseases.