Protective Effect And Mechanism Of Paeonol Derivative DPF-6 On Acute Liver Injury

Acute liver injury(ALI)is a disease that causes rapid loss of liver function accompanied by severe inflammation.The disease occurs quickly,the condition is critical,the hepatocyte are seriously damaged,and even other systems in the body can be harmed.There are many factors that can cause acute liver injury clinically,including drug-induced,excessive intake of alcohol,viral infections,and misuse of toxic foods.At present,there are no specific drugs and methods for the treatment of acute liver injury,but the inflammatory response is thought to aggravate the development of acute liver injury.Perhaps controlling the inflammatory response in acute liver injury may be one of the effective ways to protect acute liver injury.The zinc finger E-box binds to the homeobox 2(ZEB2)as an important regulator in the epithelial-mesenchymal-transformation(EMT)process,and is also a key regulator of differentiation and developmental events.Currently ZEB2 and human tumors relationship between them has been widely reported.By regulating the expression of E-cadherin,the transformation of tumor cells from epithelial phenotype to interstitial cell phenotype is promoted,and the adhesion between cells is reduced,thereby obtaining invasion.And the ability to migrate,and eventually transferred to other tissues and organs,tumor cells can regain the epithelial-like phenotype when they reach a new location,which plays a role in the development of various malignant tumors such as rectal cancer,stomach cancer,and nasopharyngeal carcinoma.An important facilitating role.In addition,ZEB2 can accelerate the proliferation of tumor cells by regulating Cyclin D1,Rb and other genes.Studies have shown that when the expression of ZEB2 is disturbed,the cell cycle G1 to S phase is blocked,and the cells proliferate,migrate and invade in vitro.The ability has dropped significantly.According to a recent study,ZEB2 regulates FcεRI-induced activation of NFAT in mast cells,where NFAT controls a variety of cellular processes and transcription of interleukins and TNF-α,which is critical for FcεRI activation-induced mast cell survival,and In the early stage,we also found that ZEB2 can inhibit nuclear factor-kappaB(NF-κB)signaling through degradation and phosphorylation of IκB kinase in acute kidney injury,resulting in a significant decrease in inflammatory cytokine levels.In this study,we investigated the protective effect and possible mechanism of paeonol derivative DPF-6 on acute liver injury in LPS/D-Gal mouse model of acute liver injury and LPS-stimulated L-02 cell model.To this end,we carry out the following experimental research(1)Protective effect of paeonol derivative DPF-6 in acute liver injury in animalsIn the acute liver injury of LPS/D-Gal combined stimulation,the paeonol derivative DPF-6 was administered,and the pathological changes of liver tissue were observed by HE staining,and the serum level of ALT/AST was detected by the kit.And the expression of inflammatory factors and ZEB2 was detected by qRT-PCR.The results showed that compared with the model group,paeonol derivative DPF-6 reduced serum ALT/AST levels at 25,50,and 100 mg/kg concentrations,improved histopathological changes,and inhibited TNF-α,IL-6,and IL.-1β secretion and promotes the expression of liver ZEB2.(2)Effect of paeonol derivative DPF-6 on expression of inflammatory cytokines and ZEB2 in LPS-induced L-02 cellsThe paeonol derivative DPF-6 was administered at concentrations of 25,50 and 100 μM to intervene LPS-stimulated L-02 cell inflammation model.The expression of inflammatory factors and ZEB2 was observed by qRT-PCR and Western blot.The results showed: compared with the LPS stimulation group,paeonol The derivative DPF-6 inhibited the secretion of TNF-α,IL-6 and IL-1β at the concentrations of 25,50 and 100 uM,and promoted the expression of ZEB2.(3)Construction of ZEB2 overexpression plasmid by genetic recombinationRecombinant eukaryotic expression vector pEGFP-C2-ZEB2 was constructed by gene recombination technology,and the gene was sequenced and transfected into 293 T cells and detected by Western blot.The results showed that the gene was sequenced and found in GencBank.The sequences were identical and it was found that the expression of ZEB2 protein was observed in 293 T cells transfected with pEGFP-C2-ZEB2 recombinant plasmid.(4)The influence of ZEB2 on the possible mechanism of inflammatory factor secretion and its influence on JNK pathwayIn LPS-stimulated L-02 cell inflammatory model,pEGFP-C2-ZEB2,pEGFP-C2-vector,siRNA-ZEB2 and siRNA-NC were used for transfection,and JNK signal in inflammatory cell model was detected by qRT-PCR and Western blot.The results showed that inhibition of ZEB2 further promoted JNK activation and secretion of inflammatory factors compared with LPS-stimulated group,and overexpression of ZEB2 inhibited JNK activation and secretion of inflammatory factors.The above results indicate that paeonol derivative DPF-6 may regulate the activation of JNK signaling pathway by regulating the expression of ZEB2,thereby inhibiting the secretion of inflammatory factors,thereby protecting acute liver injury.