PMC

Acteoside treatment on RAW 264.7 cells. (A) RAW264.7 cells were incubated without MCSF (30 ng/mL) and RANKL (25 ng/mL). (B) RAW264.7 cells were incubated with MCSF (30 ng/mL) and RANKL (25 ng/mL) for 3 days (no treatment with acteoside). (C) RAW264.7 cells were incubated with MCSF (30 ng/mL) and RANKL (25 ng/mL) for 6 days (no treatment with acteoside). (D) Effects of acteoside on the cell viability of RAW264.7, cells were treated with the indicated concentrations of acteoside for the indicated times. Cell viability was determined by the MTT method.

Acteoside does not affect L929 mouse fibroblast cell and primary rat chondrocyte viability. (a) Acteoside did not affect the viability of L929 mouse fibroblast cells. (b) Acteoside did not decrease the viability of primary rat chondrocytes. Mouse fibroblast cell line L929 used as normal cells and primary rat chondrocytes were treated with 2.5, 5, 10, 25, 50, and 100 μM acteoside for 24 h. Thereafter, MTT assay was performed to assess the cytotoxicity of acteoside in L929 cells and primary rat chondrocytes. (c) Acteoside did not increase the cytotoxicity in both L929 and primary rat chondrocytes. To perform the Cell Live/Dead assay, L929 and primary rat chondrocyte were treated with 50 and 100 μM acteoside for 24 h. Thereafter, Cell Live/Dead assay was performed. Stained cells were imaged using a fluorescence microscope (Eclipse TE200; Nikon Instruments, Melville, NY).

Upper: the molecular structure of acteoside (left) and the crystallized ligand (right). Lower: the docked complex structure of caspase-3 and acteoside with reference to the native ligand. The protein is shown in blue. Acteoside is in pink and the native ligand is in cyan. The residues forming strong interactions are in yellow. The yellow dashed lines represent the hydrogen bonds between the protein residues and bound ligands. The numbers beside the dashed lines are the distance (in Å) between the two heavy atoms of the hydrogen bonds.

Acteoside identification/quantification in mouse plasma and grafted tumors with the use of UHPLC-MS analyses. (A) Calibration curve of the analyte/IS vs. increasing concentrations of acteoside. (B) Representative LC-MS chromatogram of acteoside in mice plasma. (C) Representative MS spectrum of acteoside in treated mice plasma (C₁) and relative quantification (C₂) after OR or IP administration of the compound. (D) Quantification of acteoside in excised tumors after OR or IP administration of the compound in mice. Bars, ± SD (n ≥ 2). * , P < 0.05; * *, P < 0.01.