Bromelain is a mixture of proteolytic enzymes derived from pineapple (Ananas comosus) fruit and stem possessing several beneficial properties, particularly anti-inflammatory activity. However, the molecular mechanisms underlying the anti-inflammatory effects of bromelain are unclear. This study investigated the anti-inflammatory effects and inhibitory molecular mechanisms of crude and purified rhizome bromelains on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. RAW264.7 cells were pre-treated with various concentrations of crude bromelain (CB) or purified bromelain (PB), and then treated with LPS. The production levels of pro-inflammatory cytokines and mediators, including nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α were determined by Griess and ELISA assays. The expressions of inducible nitric oxide synthetase (iNOS), cyclooxygenase (COX)-2, nuclear factor kappa B (NF-κB), and mitogen-activated protein kinases (MAPKs)-signaling pathway-related proteins were examined by western blot analysis. The pre-treatment of bromelain dose-dependently reduced LPS-induced pro-inflammatory cytokines and mediators, which correlated with downregulation of iNOS and COX-2 expressions. The inhibitory potency of PB was stronger than that of CB. PB also suppressed phosphorylated NF-κB (p65), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha, extracellular signal-regulated kinases, c-Jun amino-terminal kinases, and p38 proteins in LPS-treated cells. PB then exhibited potent anti-inflammatory effects on LPS-induced inflammatory responses in RAW264.7 cells by inhibiting the NF-κB and MAPKs-signaling pathways.

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Rhinacanthus nasutus (Linn.) Kurz. is a medicinal plant traditionally used in northern Thailand. It has anti-bacterial, anti-inflammatory, anti-cancer, and anti-viral properties. This study investigated the anti-herpes simplex virus type 2 (anti-HSV2) activity of stem extracts of R. nasutus (Linn.) Kurz. on American green monkey kidney (or Vero) cells. The plant extracts were prepared with six different solvents: ethanol, ethylacetate, methanol, dichloromethane, acetone and hexane. The cytotoxicities of the stem extracts were determined using MTT assay with various concentrations of the stem extracts (2.44 to 156 µg/mL) and the 50% cytotoxicity dose (CD 50) was calculated after the Vero cells were treated with the stems extracts for 24 h. The methanolic extract showed a low cytotoxicity on the Vero cells, with a CD 50 value of 78.22 ± 23.54 µg/mL; in contrast, the ethylacetate, dichloromethane, acetone and hexane extracts were highly toxic to the Vero cells. The anti-HSV2 activities of the plant extracts were investigated by plaque reduction assay after treating the infected Vero cells for 24 h with the stem extracts at concentrations between 2.44 and 19.50 µg/mL. The inhibitory effect values of the 50% effective dose (ED 50) were determined. The methanolic extract had the highest inhibitory effect (ED 50 of 16.16 + 10.83 µg/mL) against HSV-2 infections and inhibited HSV-2 particles by 70.5% after 24 h of

Introduction: This research aims to evaluate the anti-diabetic activity of the extracts from different parts of Cratoxylum formosum subsp. formosum.Methods: The in vitro inhibitory activities of the hexane (HEX), dichloromethane (DCM) and ethyl acetate (EtOAc) extracts from the flowers, leaves, roots and stems on pancreatic α-amylase (pAA), Saccharomyces α-glucosidase (SAG), rat intestinal maltase (rIM), and sucrase (rIS) were investigated. Results: The DCM and EtOAc extracts from the flowers (IC50 5.4 ± 1.5 and 10.5 ± 0.6 µg/mL) displayed the similar inhibitory activities as acarbose (IC50 7.2 ± 0.4 µg/mL) in the pAA assay. The inhibitory activities of the DCM and EtOAc extracts from the flowers (IC50 56.7 ± 8.9 and 20.4 ± 0.4 µg/mL), EtOAc extract from leaves (IC50 45.0 ± 3.5 µg/mL), DCM and EtOAc extracts from roots (IC50 35.0 ± 6.7 and 16.7± 3.6 µg/mL), and EtOAc extract from stems (IC50 31.1 ± 7.3 µg/mL) were more potent than acarbose (IC50 431.4 ± 16.7 µg/mL) on SAG inhibitory assay (P<0.05). In the rIM assay, DCM and EtOAc extracts from the flowers (IC50 8.5 ± 0.2 and 12.4 ± 0.3 µg/mL) exhibited stronger inhibitory activity than acarbose (IC50 38.5 ± 7.2 µg/mL) (P<0.05). Moreover, the inhibitory activity of DCM extract from the flowers (IC50 16.9 ± 1.5 µg/ mL) was comparable to the acarbose (IC50 15.5 ± 1.2 µg/mL) on rIS assay. Conclusion: The DCM and EtOAc extracts from the flowers were more active than the leaves, roots and stems in the inhibition of our defined target enzymes.

Eucalyptus essential oils are used as traditional medicines in many countries. The objective of this study was to evaluate the antioxidant and anti-inflammatory activities of leaf essential oils extracted from three different Eucalyptus species on HepG2 and RAW264.7 cells. Essential oils were distilled from fresh leaf samples, and the chemical constituents were analyzed using gas chromatography–mass spectrometry. The antioxidant activities of essential oils were determined using ABTS and hydroxyl radical scavenging assays, and hydrogen peroxide (H2O2)-induced oxidative stress in HepG2 cells. Additionally, lipopolysaccharide (LPS)-activated RAW264.7 macrophages were used to evaluate the anti-inflammatory properties. The results revealed that Eucalyptus citriodora and Eucalyptus urophylla leaf essential oils had a high content of oxygenated monoterpenes, whereas Eucalyptus deglupta contained a high amount of monoterpene hydrocarbons. Essential oils extracted from the three Eucalyptus species showed antioxidant and anti-inflammatory activities. E. citriodora and E. urophylla leaf essential oils had strong antioxidant activity against H2O2-induced oxidative stress in human HepG2 cells. Additionally, E. citriodora leaf essential oil, which contains a high amount of citronellal, exhibited the most potent anti-inflammatory activity in LPS-activated RAW264.7 macrophages. The antioxidant and anti-inflammatory effects of essential oils depended on their chemical composition. A principal component analysis explained 100% of the variance was performed to construct three groups based on the chemical components and antioxidant and anti-inflammatory activities. This study suggests that E. citriodora leaf essential oil, which represents a good source of oxygenated monoterpenes, could be considered a potential phytochemical agent for the prevention of oxidative stress and inflammation. Keywords: Antioxidant, Anti-inflammatory effect, Eucalyptus essential oils, Gas chromatography–mass spectrometry, Principal component analysis