Figure 6.

Aloin inhibits LPS-induced apoptotic cell death. (A) RAW 264.7 cells were pre-treated with or without aloin (400 μM) for 2 h, and then stimulated with LPS (100 ng/mL) for 24 h or 48 h. Cells were collected and stained with Annexin V/PI, the percentage of Annexin-V positive cells was analyzed by flow cytometry analysis Data shown are the means ± SD of three experiments; (B) The protein expression levels of pro-caspase 9, cleaved caspase-9, pro-caspase 3, and cleaved caspase 3 were determined by immunoblotting analysis. β-actin was used as a loading control; (C) Aloin prevented the LPS-induced inflammatory response by inhibiting NF-κB signaling. Aloin attenuated LPS-induced p65 post-translational modifications by inhibiting p38 and MSK1-mediated phosphorylation and p300-mediated acetylation. This in turn prevented p65 nuclear translocation, and downregulated NF-κB mediated gene expression, including pro-inflammatory cytokines, and genes involved in apoptosis.