Chemical profiling and identification of anti-inflammatory biomarkers of oriental Thuja (Platycladus orientalis) using UPLC/MS/MS and network pharmacology-based analyses.

Platycladus orientalis L. Franco has many folk uses as it is mainly used to treat inflammatory ailments. UPLC-MS/MS was used for the chemical profiling of P. orientalis leaves. Identified metabolites were forwarded to network pharmacology analysis. Networks were constructed based on STITCH, SEA, DAVID, KEGG and STRING databases and using Cytoscape. The identified hit compounds were afzelin, myricetin, apigenin-7-O-hexoside, quercetrin and hyperoside. IL2, VEGFA, AKT1, AKT2, CREB1, IL5, RPS6KB1 and TNF were the main inflammation-related targets identified. Quercetrin and hyperoside were tested for their anti-inflammatory activity. it can be concluded that, the identified hit compounds exhibited strong synergistic interactions with the inflammation and immunity-related targets and pathways.

Seasonal dynamics of the phenolic constituents of the cones and leaves of oriental Thuja (Platycladus orientalis L.) reveal their anti-inflammatory biomarkers.

In this study, the seasonal dynamics of the flavonoids in the cones and leaves of oriental Thuja (Platycladus orientalis L. Franco) as well as the in vitro anti-inflammatory activity of their extracts were investigated. The important chemical markers of the studied extracts were determined using untargeted HPTLC profiling, which was further utilized to assess the seasonality effect on the composition of these metabolites over three seasonal cycles. A quantitative HPTLC method was developed and validated for the identified chemical markers of oriental Thuja: hyperoside, quercetrin, isoscutellarein-7-O-ß-xyloside, cupressuflavone, hinokiflavone, sotetsuflavone and isoscutellarein-8-methyl ether. The highest amounts of flavonoids were observed during the summer and winter seasons, where the leaves possessed higher contents of flavonoids compared to cones. Flavone glycosides are a major class of flavones encountered in leaves, while the cones mainly accumulated biflavones. The results showed that the effect of seasonal variation on the accumulation of flavonoids within the cones was less pronounced than in the leaves. The summer leaves showed a remarkable reduction in the levels of INF-γ, where the value decreased to 80.7 ± 1.25 pg mL-1, a significantly lower level than that obtained with piroxicam (180 ± 1.47 pg mL-1); this suggests a noteworthy anti-inflammatory potential. OPLS (orthogonal projection to latent structures) models showed that flavonoidal glycosides, quercetrin, hyperoside and isoscutellarein-7-O-ß-xyloside were the most contributing biomarkers to the reduction in pro-inflammatory mediators in LPS-stimulated WBCs. The results obtained in the study can thus be exploited to establish the best organs as well as the optimal periods of the year for collecting and obtaining certain biomarkers at high concentrations to guarantee the efficacy of the obtained extracts.

Efficacy-directed discrimination of the essential oils of three Juniperus species based on their in-vitro antimicrobial and anti-inflammatory activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Juniperus plants are considered important sources of cedar-wood oil which is used widely in folk medicine as antiseptic and in treatment of inflammatory disorders such as, rheumatoid arthritis but there is not enough scientific evidence to support the claimed uses and there is no specification of a certain Juniperus species as the most active. AIM OF THE STUDY: The aim of this study is volatiles profiling of three Juniperus species; J. communis, J. horizontalis and J. chinensis in addition to efficacy-directed discrimination of the three studied essential oils based on their antimicrobial, and anti-inflammatory activities in LPS (lipopolysaccharide)-stimulated WBCs (White blood cells) to investigate the inter-specific variability effect on the biological activities of each oil. MATERIALS AND METHODS: Volatile components profiling of the three studied plants volatile oils was achieved using GC-FID (Gas chromatography - flame ionization detector) and GC-MS (Gas chromatography - mass spectrometry). The antimicrobial activity of the studied essential oils was investigated and the minimum inhibitory concentration (MIC) was determined for oils. The production of the pro-inflammatory cytokines was evaluated by ELISA (Enzyme linked immunosorbent assay). Identification of the biomarkers responsible for each activity was attempted through construction of orthogonal projection to latent structures model using multivariate statistical analysis. RESULTS: Forty five components were identified in the volatile oils of the three studied plants. J. horizontalis oil displayed the highest activity against E. coli while J. communis showed the highest activity against S. aureus. OPLS model biplot showed the in-between class discrimination of J. chinensis oil sample from J. communis and J. horizontalis. The three oils were found to significantly decrease the production of the pro-inflammatory cytokines tumour necrosis factor (TNF)- α, interleukin (IL)-1ß, and gamma interferon (INF- γ) in lipopolysaccharide-activated white blood cells. All studied oils were similar in reduction of TNF-α, and INF-γ, while J. chinensis oil possessed the highest potency against IL-1ß. The coefficient plots of TNF-α and INF-γ pro-inflammatory mediators showed that 1-terpineol, 4-terpineol, bornyl acetate, dl-limonene and α-pinene positive contributors to both activities while ß-thujone, 3-carene and γ-muurolene were the positive contributors to IL-1ß inhibitory activity. CONCLUSION: The differences observed in the volatile profiles among the three studied oils demonstrate the effect of inter-specific variability on the biological activities of the tested oils. It was shown that the tested oils possessed good antibacterial activities against E.coli and S. aureus justifying its folk use as an a topical antiseptic while the observed anti-inflammatory effects in human WBCs is due at least in part to their inhibitory effect on the production of pro-inflammatory cytokines.