Aloperine

232, 36

Female BALB/c mice with DNFB-induced allergic contact dermatitis

Dissolved in DMSO, final concentrations ca.2 mM

Mixed in the emollient cream vehicle

Topical application of 1% Aloperine suppresses 2, 4-dinitrofluorobenzene (DNFB)-induced increase in ear thickness and ear erythema in BALB/c mice, and significantly decreases the up-regulated mRNA and protein levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) induced by DNFB in ear biopsy homogenates. [1] Topical application of Aloperine reduces the DNFB-induced dermatitis (dermatitis index and ear thickness) in NC/Nga mice at day 13 and day 14 in a dose-dependent manner. Aloperine treatment reduces DNFB-induced lymphocytes infiltration and eosinophils infiltration in a dose-dependent manner. Aloperine treatment also reduces the DNFB-induced infiltration of mast cells in a dose-dependent manner., Aloperine reduces the plasma level of IgE in a dose-dependent manner. Aloperine markedly reduces DNFB-induced increase in IL-4, IL-13 and IFN-gamma productions, while it increases the IL-10 level in a dose-dependent manner. Aloperine treatment significantly reduces the cytokine levels of TNF-alpha, IL-1beta and IL-6 in ear biopsies homegenates of NC/Nga mice in a dose-dependent manner. [3]

Cells are seeded in 96-well plates in 100-uL culture medium. After incubation of 4 hours for leukaemia cells and of 24 hours for solid cancer cells, experimental media containing either excipient control or Aloperine are added to appropriate wells. Five concentrations of Aloperine for 48-hour treatment are used to determine the in vitro IC50 growth inhibitory values of Aloperine in cancer cells. After incubation, 10 uL of MTT solution (5 mg/mL) is added to each well. The plates are then incubated for 4 hours at 37 C. Intracellular formazan crystals are dissolved by addition of 100 uL of isopropanol-HCI-SDS solution to each well. After an overnight incubation at 37 C, the optical density of the samples is determined at 570 nm. DNA fragmentation is analysed after the extraction of DNA from cells exposed to the indicated doses of Aloperine for 48 hours using apoptotic DNA ladder kit. For autophagy detection, cells are collected and incubated with PBS containing 5 uM acridine orange for 15 minutes. The acridine orange is removed and the cells are resuspended in 100 uL of PBS. Fluorescent micrographs are obtained with an inverted fluorescent microscope. Autophagy is quantified based on the mean number of cells displaying intense red staining for three fields (containing at least 50 cells per field) for each experimental condition.

Aloperine is an isolated alkaloid in sophora plants such as Sophora alopecuroides L, and exhibits anti-inflammatory, antibacterial, antiviral, and anti-tumor properties.