Mechanism Of Chlorogenic Acid Attenuates Sepsis Via PI3K/AKT Pathway

Part Ⅰ Mechanism of chlorogenic acid in reducing inflammation in animal models[Objective] To investigate the effect of chlorogenic acid on inflammatory response and its possible signal pathway in vivo.[Methods]Male Balb/c mice were used as experimental animals.The animals were divided into blank group,model group,experimental group and positive drug control group.Sepsis model was established by intraperitoneal injected LPS.Eye blood was collected to measure the level of ALT and AST.The liver tissue of mice was stained by HE,and the degree of liver injury was observed.The levels of IL-6,TNF-αand INOS were detected by RT-PCR and PI3K / AKT signal pathway were detected by West-Blot.[Results]Compared with the blank control group(N Group),the level of ALT and AST in the model group(LPS group)increased significantly(p < 0.05).The level of Proinflammatory cytokine increased significantly(p < 0.05)and the structural disorder of hepatic lobules was observed in HE section of liver which filled with inflammatory cells.The protein expression of PI3K / AKT was also significantly increased(p < 0.05).Compared with the model group,chlorogenic acid significantly reduced the levels of ALT and AST in vivo(p < 0.05)and the levels of inflammatory factors in cells(p < 0.05).The HE section of liver can be obviously observed decrease the disorder of tissue structure,the hyperemia of tissue and the infiltration of inflammatory cells.The protein expression of NF-κBp65 and PI3K / AKT were significantly decreased(p < 0.05).[Conclusion] In vivo experiments,chlorogenic acid can reduce the inflammatory response induced by sepsis,and PI3K / AKT Pathway may be the pathway of chlorogenic acid exerting anti-inflammatory mechanism.Part Ⅱ Chlorogenic acid attenuates inflammation via PI3K/AKT pathway[Objective] To explore the specific signal pathway of chlorogenic acid in reducing inflammation.[Methods]Using Raw264.7 cell line as experimental cells,LPS was used to construct a cell model of sepsis.chlorogenic acid was divided into high,middle and low experimental groups and dexamethasone as positive control group.To observe the entry of P65 into the nucleus immunofluorescence was used and the reactive oxygen species(Ros)by flow Quantitative analysis.LY294002 as a specific inhibitor of PI3K detect whether PI3K / Akt was the specific signal pathway of anti-inflammatory effect of chlorogenic acid.The change of PI3K / AKT signal pathway was detected by West-Blot.The anti-inflammatory effect of chlorogenic acid was studied by real-time quantitative method.[Results]Compared with the blank control group(N Group),the Model Group(LPS group)can significantly increase the active oxygen and inflammatory factors and P65 into the nucleus.Chlorogenic acid can significantly reduce the active oxygen,inflammatory factors and P65 into the nucleus.With the concentration of chlorogenic acid increased,the more obvious intervention.LY294002,as a specific inhibitor of PI3K,can obviously reduce the level of reactive oxygen species,inflammatory factors and P65 entering the nucleus,and can also reduce the expression of P65.The effect of LY294002 is similar to that of low concentration chlorogenic acid group,chlorogenic acid can reduce the expression of PI3K and AKT.[Conclusion]Chlorogenic acid can reduce P65 entry into the nucleus and the production of inflammatory factors and reactive oxygen species,which through the PI3K/ AKT pathway.