The Experimental Study Of Artesunate On TNF?-induced Inflammatory Response Of HepG2 Cells By Regulating The MAPK/NF-?B Signaling Pathways

Objective:To explore the effect of different concentration of artesunate(Art)on inflammation response of tumor necrosis factor(TNF?)-induced HepG2 cells and its mechanism of its action.Methods:HepG2 cells were resuscitated and cultured.After the cells were grown to a certain number,plating and administration were performed.The nontoxic cell concentration of artesunate was detected by MTS assay.The optimal time and concentration of TNF?-induced hepatitis model were determined by Western blot.Based on the above results,the experiments were divided into 4 groups:normal group,model group(TNF?),administration group 1(TNF? + low concentration Art)and administration group 2(TNF? + high concentration Art).Western blot was used to determine differences in the expression of(p)JNK,(p)Erk,(p)p38,pp65 and COX2 protein in different groups,and the expression differences of COX2 gene in different groups were determined by RT-PCR.Results:1.MTS results showed that with the increase of artesunate concentration,the cell survival rate decreased in a dose-independent manner.2.Compared with the blank group,the expression of COX2 protein in the high concentration(20ng/mL)TNFa group was higher than that in the low concentration(10ng/mL)group.Under the circumstance of a certain dosage concentration,the expression of COX2 protein gradually decreased over time.3.Compared with the blank group,the expression of COX2 gene and protein were significantly increased in the model group,which demonstrated that the inflammation model of hepatocellular carcinoma cells was successfully established.After the administration,the expression of the above gene and protein was significantly reduced,and this change was dose-dependent and the difference was statistically significant(P<0.01).4.Compared with the control group,the expression of pJNK,pErk,pp38,and pp65 proteins in the model group increased significantly(P<0.01).Compared with the model group,the expression of above proteins in the treatment group was obviously reduced,and this change is remarkably significant(P<0.05).Conclusion:1.TNF? can activate the related proteins(pJNK,pErk,pp38,pp65)of MAPK and NF-?B pathway in HepG2 cells,promote the expression of COX2 gene and protein,and participate in the development of inflammation.2.Artesunate can inhibit the activation of related proteins(pJNK,pErk,pp38,pp65)in MAPK and NF-?B pathway in TNF?-stimulated HepG2 cells,downregulate the expression of COX2 gene and protein,suggesting that Artesunate may have a potential anti-inflammatory effect.