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Probiotics offer various health benefits. Lactobacillus plantarum has been used for decades to enhance human intestinal mucosal immunity and improve skin barrier integrity. Extracellular vesicles (EVs) derived from eukaryotic or prokaryotic cells have been recognized as efficient carriers for delivery of biomolecules to recipient cells, and to efficiently regulate human pathophysiology. However, the mechanism underlying the beneficial effects of probiotic bacteria-derived EVs on human skin is unclear. Herein, we investigated how L. plantarum-derived EVs (LEVs) exert beneficial effects on human skin by examining the effect of LEVs on cutaneous immunity, particularly on macrophage polarization. LEVs promoted differentiation of human monocytic THP1 cells towards an anti-inflammatory M2 phenotype, especially M2b, by inducing biased expression of cell-surface markers and cytokines associated with M2 macrophages. Pre- or post-treatment with LEVs under inflammatory M1 macrophage-favouring conditions, induced by LPS and interferon-γ, inhibited M1-associated surface marker, HLA-DRα expression. Moreover, LEV treatment significantly induced expression of macrophage-characteristic cytokines, IL-1β, GM-CSF and the representative anti-inflammatory cytokine, IL-10, in human skin organ cultures. Hence, LEVs can trigger M2 macrophage polarization in vitro, and induce an anti-inflammatory phenomenon in the human skin, and may be a potent anti-inflammatory strategy to alleviate hyperinflammatory skin conditions. [ABSTRACT FROM AUTHOR]

Introduction: Camellia japonica L. (Theaceae) is an evergreen shrub, which is cultivated as a popular ornamental tree in Korea, China, and Japan and its seeds have been used as a source of cooking oil, in cosmetics and as a traditional medicine. Intensive phytochemical works have revealed that oleanane‐type saponins are the characteristic compounds of the seeds of C. japonica. Objective: The purpose of the present study is to isolate and determine oleanane‐type saponins from C. japonica using high‐performance countercurrent chromatography (HPCCC) coupled with reversed‐phase high‐performance liquid chromatography (RP‐HPLC) and spectroscopic evidences, respectively. Methodology: HPLC electrospray ionisation quadrupole time‐of‐flight mass spectrometry (ESI‐Q‐TOF‐MS) was applied to profile the saponin composition of an enriched saponin extract of C. japonica seeds. The enriched saponin extract was separated by HPCCC using a dichloromethane/methanol/isopropanol/water (9:6:1:4, v/v/v/v) system and RP‐HPLC. The structures of the isolates were determined utilising ESI‐Q‐TOF‐MS, one‐dimensional and two‐dimensional NMR and optical rotation. Results: HPCCC on enriched saponin extract of C. japonica yielded four saponin fractions in the order of the number of sugars attached to the triterpene aglycone, and preparative RP‐HPLC on each saponin fraction led to the isolation of nine novel saponins, namely camoreoside A–I, along with six known ones. Conclusions: This study indicates that combination of HPLC‐ESI‐Q‐TOF‐MS analysis and HPCCC coupled with RP‐HPLC are excellent tools for discovering saponins from natural sources. High‐performance liquid chromatography electrospray ionisation quadrupole time‐of‐flight mass spectrometry (HPLC‐ESI‐Q‐TOF‐MS) revealed that enriched saponin extract of Camellia japonica contained a variety of saponins with di‐, tri‐, tetra‐saccharides as oligo‐glycoside moieties. High‐performance countercurrent chromatography (HPCCC) coupled with reversed‐phase (RP) HPLC were successfully applied to isolate nine new saponins namely camoreoside A–I along with six known ones from enriched saponin extract of C. japonica seeds. The structures of the isolates were determined utilising ESI‐Q‐TOF‐MS, one‐dimensional and two‐dimensional NMR and optical rotation. [ABSTRACT FROM AUTHOR]

High-performance countercurrent chromatography (HPCCC) with electrospray light-scattering detection was applied for the first time to isolate a spirostanol and a novel furostanol saponin from Liriope platyphylla. Due to the large differences in KD values between the two compounds, a two-step HPCCC method was applied in this study. The primary HPCCC employed methylene chloride/methanol/isopropanol/water (9:6:1:4 v/v, 4 mL/min, normal-phase mode) conditions to yield a spirostanol saponin ( 1). After the primary HPCCC run, the solute retained in the stationary phase (SP extract) in HPCCC column was recovered and subjected to the second HPCCC on the n-hexane/ n-butanol/water system (1:9:10 v/v, 5 mL/min, reversed-phase mode) to yield a novel furostanol saponin ( 2). The isolated spirostanol saponin was determined to be 25( S)-ruscogenin 1- O-β- d-glucopyranosyl (1→2)-[β- d-xylopyranosyl (1→3)]-β- d-fucopyranoside (spicatoside A), and the novel furostanol saponin was elucidated to be 26- O-β- d-glucopyranosyl-25( S)-furost-5(6)-ene-1β-3β-22α-26-tetraol-1- O-β- d-glucopyranosyl (1→2)-[β- d-xylopyranosyl-(1→3)]-β- d-fucopyranoside (spicatoside D). [ABSTRACT FROM AUTHOR]

A rapid and efficient high-performance counter-current chromatography ( HPCCC) method was developed to separate five oligostilbenes from the roots of Vitis amurensis. An n-hexane/ethyl acetate/methanol/water system (4:8:4:10, v/v/v/v) was selected as an optimal two-phase solvent system of which the upper phase was used as the stationary phase and the lower phase was used as the mobile one. Partition coefficient values for the target compounds under these optimized conditions were 0.28 ( 1, ampleosin A), 7.12 ( 2, (+)- g-viniferin), 2.26 ( 3, vitisin A), 5.38 ( 4, wilsonol C), and 11.23 ( 5, vitisin B). Flow-rate gradient HPCCC (4 mL/min in 0-70 min, 8 mL/min in 70-250 min) was applied to isolate the target compounds in as high purity as possible within the shortest possible run time. Under these conditions, ampelopsin A (12.1 mg), (+)- g-viniferin (10.4 mg), vitisin A (2.8 mg), wilsonol C (3.2 mg), and vitisin B (37 mg) were isolated with >95% purity from 150 mg of enriched oligostilbene extract. Although the KD of the last eluted compound, vitisin B ( KD = 11.23), was relatively large, it was eluted in 115-145 min using the two-phase solvent system. This study shows that HPCCC is an efficient tool for the isolation and purification of natural products. [ABSTRACT FROM AUTHOR]