The Anti-inflammatory Effect And Mechanism Of Tubeimoside Ⅲ By Mediating Glucose Metabolism Reprogramming Of Macrophages

Objective: To explore the anti-inflammatory effect of tubeimoside Ⅲ in sepsis,and to study its anti-inflammatory mechanism.Research methods:1.C57BL/6 mice were injected intraperitoneally with lipopolysaccharide(LPS)to establish an acute sepsis model.Tissue sections,HE staining,and RT-PCR were used to detect the effect of tubeimoside Ⅲ on the inflammatory response in mice.2.NO production was detected by Griess method,and the expression changes of various inflammatory factors were detected by RT-PCR and Elisa,so as to clarify the anti-inflammatory effect of tubeimoside Ⅲ in LPS-induced inflammatory macrophages.3.In LPS-induced RAW264.7 macrophage model,Western Blot,energy metabolism analysis by Seahorse,pyruvate content by colorimetric detection,RT-PCR and other experiments were used to explore the molecular mechanism of the antiinflammatory effect of tubeimoside Ⅲ.The results were verified by inhibition in LPSactivated RAW264.7 macrophages and in a mouse model of sepsis.Research results:1.In acute sepsis model mice,tubeimoside Ⅲ can reduce inflammation and tissue damage in the organs of mice,inhibit m RNA expression levels of IL-1β,IL-6 and i NOS,and play an anti-inflammatory role.2.In RAW264.7 macrophages induced by LPS,Tubeimoside Ⅲ could effectively inhibit the expression of IL-1β,IL-6 and i NOS,but had no significant effect on the m RNA expression levels of TNF-α,COX-2,CXCL15,HMGB1 and GM-CSF.3.In LPS-induced RAW264.7 macrophages,Tubeimoside Ⅲ reduced the production of lactic acid and oxidative phosphorylation,and increased the production of pyruvate.At the same time,the expression of key rate-limiting enzyme genes HK2,GLUT1 and PFKFB3 in glycolytic metabolism pathway was up-regulated,while the LDHA gene was inhibited,indicating that Tubeimoside Ⅲ regulates the glucose metabolism reprogramming of macrophages.In addition,Tubeimoside Ⅲ played a similar regulatory effect on downstream effector molecules of itaconic acid.Through using itaconate inhibitor in RAW264.7 macrophage and sepsis mouse model,it was proved that Tubeimoside Ⅲ could inhibit the expression of IL-1β and IL-6,and its antiinflammatory effect and organ protection required the involvement of itaconic acid.Conclusion: Tubeimoside Ⅲ can inhibit the expression of IL-1β and IL-6 in inflammatory macrophages,and play an anti-inflammatory role,which is realized through itaconate and its downstream signaling pathway.