Phenolic acids as chemotaxonomic markers able to differentiate the Euphrasia species.

The objective of this study was to get the answer on the question, which phenolic compounds may serve as chemical markers in taxonomy of Euphrasia spp. (E. nemorosa, E. rostkoviana, E. stricta) collected from the wild. Moreover, it is still unknown how and which environmental parameters can impact on the quantity of polyphenols, flavonoids and phenolic acids. To the authors' knowledge, this is the first study that analyses such a broad spectrum of phenolic compounds in the genus Euphrasia and takes into account the habitat conditions of their synthesis. The species of Euphrasia differ significantly in the content of phenolic compounds. Euphrasia rostkoviana has the highest total content of polyphenols and flavonoids. Euphrasia stricta contains the largest amounts of phenolic acids, such as 5-O-caffeoylquinic, p-coumaric, protocatechuic and salicylic acid. It has been evidenced that the content of metabolites in the Euphrasia herb depends on a number of habitat parameters. The increased content of phenolic acids in E. stricta can be attributed to the fact that this species occupies drier habitats, with lower content of nitrogen and organic carbon compared to E. nemorosa and E. rostkoviana. The compounds that can be considered as chemotaxonomic markers are salicylic and protocatechuic acid for E. stricta, 5-O-caffeoylquinic acid for E. rostkoviana and ferulic acid for E. nemorosa. These findings provide theoretical and empirical basis for a chemotaxonomic classification of those species, which taken together with morphological characteristics should prevent misidentification.

Green synthesis of gold nanoparticles using Euphrasia officinalisleaf extract to inhibit lipopolysaccharide-induced inflammation through NF-κB and JAK/STAT pathways in RAW 264.7 macrophages.

BACKGROUND: Gold nanoparticles (AuNPs) have potential applications in the treatment and diagnosis process, which are attributed to their biocompatibility and high efficiency of drug delivery. In the current study, we utilized an extract of Euphrasia officinalis, a traditional folk medicine, to synthesize gold nanoparticles (EO-AuNPs), and investigated their anti-inflammatory effects on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. MATERIALS AND METHODS: The AuNPs were synthesized from an ethanol extract of E. officinalis leaves and characterized using several analytical techniques. Anti-inflammatory activities of EO-AuNPs were detected by a model of LPS-induced upregulation of inflammatory mediators and cytokines including nitric oxide (NO), inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), IL-1ß, and IL-6 in RAW 264.7 cells. The activation of nuclear factor (NF)-κB and Janus kinase/signal transducer and activators of transcription (JAK/STAT) signaling pathways was investigated by Western blot. RESULTS: The results confirmed the successful synthesis of AuNPs by E. officinalis. Transmission electron microscopy images showed obvious uptake of EO-AuNPs and internalization into intracellular membrane-bound compartments, resembling endosomes and lysosomes by RAW 264.7 cells. Cell viability assays showed that EO-AuNPs exhibited little cytotoxicity in RAW 264.7 cells at 100 µg/mL concentration after 24 hours. EO-AuNPs significantly suppressed the LPS-induced release of NO, TNF-α, IL-1ß, and IL-6 as well as the expression of the iNOS gene and protein in RAW 264.7 cells. Further experiments demonstrated that pretreatment with EO-AuNPs significantly reduced the phosphorylation and degradation of inhibitor kappa B-alpha and inhibited the nuclear translocation of NF-κB p65. In addition, EO-AuNPs suppressed LPS-stimulated inflammation by blocking the activation of JAK/STAT pathway. CONCLUSION: The synthesized EO-AuNPs showed anti-inflammatory activity in LPS-induced RAW 264.7 cells, suggesting they may be potential candidates for treating inflammatory-mediated diseases.

Ecofriendly synthesis of silver and gold nanoparticles by Euphrasia officinalis leaf extract and its biomedical applications.

Biogenic synthesis of silver (AgNPs) and gold nanoparticles (AuNPs) using aqueous extract of Euphrasia officinalis has been reported. Stable AgNPs and AuNPs were formed on adding aqueous solutions of silver nitrate and chloroauric acid with E. officinalis leaf extract, in 19 min and 2 min, respectively. The synthesis method used in present study was simple, reliable, rapid, cost effective and ecofriendly. The synthesized nanoparticles were characterized with field emission transmission electron microscopy (FE-TEM), elemental mapping, selected area diffraction pattern (SAED), energy-dispersive X-ray spectroscopy (EDS), X-ray diffractometer (XRD), particle size distribution, zeta potential and Fourier-transform infrared spectroscopy (FTIR). The UV-Vis spectrum confirmed the synthesis of nanoparticles as the absorption band was observed at 450 nm for AgNPs and at 558 nm for AuNPs. The TEM images revealed quasi-spherical shape of AgNPs and AuNPs. The size of nanoparticles was determined to be 40.37 ± 1.8 nm for AgNPs and 49.72 ± 1.2 nm for AuNPs. The zeta potential value demonstrated the negative surface charge and stable nature of nanoparticles. Crystalline nature of the nanoparticles in the face-centred cubic (fcc) structure was confirmed by the peaks in the XRD pattern and SAED pattern. FTIR results showed the functional groups involved in reduction of silver and gold ions to metal nanoparticles. For biomedical application, the nanoparticles have been explored for anticancer, antibacterial and biofilm inhibition activities. It was observed that AgNPs exert anticancer activity against human lung cancer (A549) and human cervical cancer (HeLa) cell lines. On the other hand, AuNPs were able to inhibit only human cervical cancer cells. Furthermore, the AgNPs were active against clinically isolated human pathogens like Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Vibrio parahaemolyticus. Additionally, AgNPs also showed biofilm inhibition activity against S. aureus and P. aeruginosa.

Pharmacological activities of an eye drop containing Matricaria chamomilla and Euphrasia officinalis extracts in UVB-induced oxidative stress and inflammation of human corneal cells.

Ultraviolet B (UVB) exposure is a risk factor for corneal damage resulting in oxidative stress, inflammation and cell death. The aim of this study was to investigate the potential protective effects of a commercial eye drop (Dacriovis™) containing Matricaria chamomilla and Euphrasia officinalis extracts on human corneal epithelial cells (HCEC-12) against UVB radiation-induced oxidative stress and inflammation as well as the underlying mechanisms. The antioxidant potential of the eye drops was evaluated by measuring the ferric reducing antioxidant power and the total phenolic content by Folin-Ciocalteu reagent. HCEC-12 cells were exposed to UVB radiation and treated with the eye drops at various concentrations. Cell viability, wound healing assay, reactive oxygen species (ROS) levels, protein and lipid oxidative damage and COX-2, IL-1ß, iNOS, SOD-2, HO-1 and GSS gene expression, were assessed. Eye drops were able to protect corneal epithelial cells from UVB-induced cell death and ameliorated the wound healing; the eye drops exhibited a strong antioxidant activity, decreasing ROS levels and protein and lipid oxidative damage. Eye drops also exerted anti-inflammatory activities by decreasing COX-2, IL-1ß, iNOS expression, counteracted UVB-induced GSS and SOD-2 expression and restored HO-1 expression to control levels. These findings suggest that an eye drop containing Matricaria chamomilla and Euphrasia officinalis extracts exerts positive effects against UVB induced oxidative stress and inflammation and may be useful in protecting corneal epithelial cells from UVB exposure.