Oridonin regulates the polarized state of Kupffer cells to alleviate nonalcoholic fatty liver disease through ROS-NF-κB

• Ori suppressed ROS production and regulated the M1 polarization of KCs, thus protecting the liver in NAFLD. • Regulating the M2 polarization of KCs can inhibit NAFLD inflammation and promote liver recovery. • ROS is an important signal that regulates the M2 polarization of KCs cells. Oridonin (Ori) is a kind of diterpenoid small molecule, but its role in nonalcoholic fatty liver disease (NAFLD) has not been reported yet. This study aimed to explore the pharmacological function of Ori in liver protection through the reactive oxygen species (ROS)-mediated polarization of Kupffer cells (KCs). In the present work, KCs were adopted for study in vitro. To be specific, LPS and IFN-γ were utilized to induce M1 polarization, then the influence of Ori intervention on the expression of inflammatory factors IL-1β, IL-6 and TNF-α was detected by enzyme-linked immunosorbent assay (ELISA), that of CD86 and P 65 was measured through fluorescence staining, that of p- P 65 and p-P50 was detected by Western blotting (WB) assay, and ROS expression was measured by using the DCFH-DA probe. The C57BL/6J mice were fed with the high fat diet (HFD) to construct the NAFLD model, and intervened with Ori. The blood glucose (BG), body weight (BW), food intake and water intake of mice were monitored; meanwhile, glucose and insulin tolerance tests were conducted. The liver tissues of mice were subjected to H&E staining and oil red O staining. Moreover, the serum ALT, AST and TG levels in mice were monitored, the CD86 and CD206 levels were measured through histochemical staining, the expression of inflammatory factors was detected by ELISA, and the p- P 65 and p-P50 protein levels were detected by WB assay. Ori suppressed the M1 polarization of KCs, reduced the levels of inflammatory factors, and decreased the expression of ROS, p- P 65 and p-P50. In animal experiments, Ori improved lipid deposition and liver injury in the liver tissues of NAFLD mice, increased the proportion of M2 cells (up-regulated CD206 expression), reduced that of M1 cells (down-regulated CD86 expression), and decreased the serum ALT, AST and TG levels. This study discovered that Ori suppressed ROS production and regulated the M1 polarization of KCs, thus protecting the liver in NAFLD.