Inhalation of panaxadiol alleviates lung inflammation via inhibiting TNFA/TNFAR and IL7/IL7R signalling [Human]
In order to study the effects and mechanisms of PD inhalation drug delivery, models of LPS-induced murine holistic pneumonia, isolated macrophage inflammation, and epithelial cell co-culture are utilized. For the evaluation of efficacy, pathology and molecular assays have been utilized. Mechanisms of action and targets were screened and validated by molecular means using transcriptome sequencing. Efficacy and mechanism of action were ultimately validated using human BALF cell models. Also studied were pharmacokinetic parameters of inhaled PD. Overall design: Human monocytic THP-1 and A549 were obtained from the Shanghai Institute of Cell Biology (Shanghai, China). THP-1 cells were cultured in RPMI 1640 (GIBCO, Grand Island, NY) containing 10% FBS (GIBCO, Grand Island, NY) in 5% CO2 at 37 °C. Before further treatment, THP-1 cells (1×106 cells/mL) were inoculated into 6-well plates and treated with 500 nM PMA for 4 h to induce macrophage differentiation. Next, cells were washed 3 times with phosphate-buffered saline (PBS) and incubated for 24 h to exclude PMA interference. THP-1 cells treatment was divided into 3 groups: a control group with no treatment, a model group with LPS at 1 µg/mL, a PD group with 1 µg/mL of LPS and 10 µM of PD.