Oridonin protects LPS-induced acute lung injury by modulating Nrf2-mediated oxidative stress and Nrf2-independent NLRP3 and NF-κB pathways

Ori inhibited cytotoxicity and ROS generation in RAW 264.7 cells. a The chemical structure of Oridonin (Ori). b Cells were exposed to Ori (2.5, 5 or 10？μM) for 1？h and then trea ted with or without hydrogen peroxide (300？μM) for another 18？h. Cell viability was determined by MTT assay. c Cells were treated with or without Ori for 18？h, stained with 50？μM of DCFH-DA for 30？min, and subsequently exposed to hydrogen peroxide (300？μM) for 10？min to induce ROS generation. DCF fluorescence intensities were detected by a fluorescence microscope. d and e According to the related instructions, LAL enzyme and LPS-LBP-binding assay were measured by the different absorbance. f and g Cells were exposed to Ori (2.5, 5 or 10？μM) for 1？h, treated with or without LPS (1？μg/ml) for another 18？h and stained with 50？μM of DCFH-DA for 30？min. DCF fluorescence intensities were detected by flow cytometry. All results were expressed as the means ± SEM of three independent experiments. *p？<？0.05 and **p？<？0.01 versus the control group, #p？<？0.05 and ##p？<？0.01 versus LPS grou