The Role Of Hyperoside Regulates Macrophages Polarization In Non-alcoholic Fatty Liver Disease

Background Non-alcoholic fatty liver disease(NAFLD)has become one of the most significant liver diseases and is occurring at epidemic levels worldwide.To date,no medications for NAFLD have been approved by relevant governmental agencies.Emerging evidence indicates that innate immune mechanisms are pivotal drivers of inflammation and other pathological manifestations observed in NAFLD.Hyperoside is a flavonoid compound mainly found in medicinal plants that has many biological effects,such as anti-inflammatory,antioxidative,and vascular protective effects.Hyperoside has also been shown to suppress inflammatory cytokine production in macrophages,although the role of hyperoside in the physiological process of NAFLD is poorly defined.Objective Therefore,the present study aimed to determine the role of hyperoside in high-fat diet(HFD)-induced NAFLD in mice and to investigate the molecular mechanisms underlying the effects of hyperoside.Methods 1.The wild-type C57BL/6 mice were put on a diet with HFD to building a NAFLD model.We obtained liver sections for hematoxylin and eosin(HE)staining,Oil Red O staining,Sirius red staining,to visualize hepatosteatosis and fibrosis,GTT and ITT were used to assess glucose tolerance and insulin sensitivity in mice;2.Detection of macrophage infiltration in liver and epididymal adipose tissue of NAFLD mice by immunohistochemical staining,real-time quantitative PCR(q PCR)analysis of the expression of inflammatory cytokines in liver tissues,and Flow cytometry be used to analysis peripheral blood mononuclear macrophage subpopulations(M1,M2)differentiation;3.Mouse bone marrow-derived macrophages(BMDMs)are cultured in vitro,q PCR evaluates the effect of Hyperoside on the expression of macrophage subpopulation markers to clarify its effect on macrophage polarization,and the expression of Nr4A1 in BMDMs and monocyte macrophages was detected by q PCR and Western blot;4.Based on the above experimental results,Nr4A1 knockout mice were used to verify the target of Hyperoside on NAFLD,and to determine whether the regulation of Hyperoside to macrophage polarization depends on Nr4A1.Results 1.Hyperoside attenuates HFD-induced wild-type mice hepatic steatosis and fibrosis;2.Hyperoside attenuates the HFD-induced wild-type mice systemic insulin resistance;3.Hyperoside blunts the HFD-induced wild-type mice macrophage-related inflammatory responses and regulate macrophage subpopulation differentiation;4.Hyperoside regulates macrophage polarization via up?regulation of the expression of Nr4A1 5.Nr4A1 deficiency counteracts the protective effect of Hyperoside on hepatic steatosis,fibrosis and insulin resistance.Conclusions This study demonstrated that hyperoside exerts protective effects against high-fat diet(HFD)-induced NAFLD and regulates macrophage polarization in an Nr4A1-dependent manner.After HFD,hepatic steatosis,insulin resistance,and inflammatory responses were significantly ameliorated in hyperoside-treated HFD-fed wild-type mice,and hyperoside facilitated the polarization of macrophages from pro-inflammatory M1 to the anti-inflammatory M2 subtype.Nr4A1 was found to be upregulated in hyperoside-treated HFD-fed mice,and hyperoside did not improve HFD-induced NAFLD or regulate macrophage polarization in Nr4A1-deficient mice.In conclusion,hyperoside may have therapeutic potential in preventing the pathological progression of NAFLD.