Study On JAK-STAT Signaling Pathway To Explore The Effect Of Modified Xiaochaihu Decoction On HepG2.2.15 Cells

Objective:Based The aim of this study was to investigate the effect of Xiaochaihu decoction on HepG2.2.15 cells and JAK-STAT signaling pathway,which could stably express hepatitis B virus protein and secrete virus particles.To clear wether the Xiaochaihu decoction play a role throught the JAK-STAT signaling pathway and effect the development of CHB,revealing the molecular mechanism of CHB in the treatment of CHB,and provide the theoretical basis for the treatment of hepatitis B and traditional Chinese medicine and western medicine,and lay the foundation for the development of anti-HBV drug.Methods:1.CHB patients included in the study were treated by the modified Xiaochaihu decoction which formulated under the treatment rule of Soothing Liver and Strengthening Spleen and Western medicine entecavir,drawing blood after patients have taken both kinds of medicine,prepared the drug-contained serum.2.HepG2.2.15 cells were cultured.3.HepG2.2.15 cells were cultured in drug-contained serum by the method of serum pharmacology.4.The effect of Xiaochaihu decoction on the growth of HepG2.2.15 cells was evaluated by the MTT assay to find out wether the Xiaochaihu decoction was effected on the HepG2.2.15 cell vibility which intervented by different drug concentration of serum and time periods.5.The HepG2.2.15 cell supernatant was collected after been intervented by different drug concentration of serum and time periods,used ELISA to detected the levels of HBsAg and HBeAg to evaluated the effect of Xiaochaihu decoction on the replication of HBV virus.6.HepG2.2.15 cells were cultrued by different drug concentrations of serum and intervented by different time periods.The expression of JAK2 and STAT3 protein in JAK-STAT signal transduction pathway was detected by Western Blot to evaluated the effect of Xiaochaihu decoction on the JAK-STAT pathways.Results:1.MTT experiment:HepG2.2.15 cells were cultrued by different drug concentrations of serum and intervented with 48h,96h,144h,the result shows that the Western medicine and traditional Chinese medicine were all had the inhibitory effect on cell viability but Western medicine were more obvious.2.ELISA test:Western medicine and traditional Chinese medicine all can achieve the effect of reducing the secretion of HBsAg and HBeAg in serum,and we could find the time-dependent in 20%-content Chinese medicine group while the 10%-content Chinese medicine group and the Western medicine group did not(With the prolongation of drug intervention time,the efficancy of Xiaochaihu decoction becoming increasingly evident).In addition to that the concentration-dependent was observed in all traditional Chinese medicine groups(and the 20%-content Chinese medicine group was found more efficancy than 10%-content group)3.Compared with the control group,the expression of JAK2/STAT3 protein in the western medicine group,10%Chinese medicine group and 20%Chinese medicine group decreased gradually,and there was significant difference between the group and the group(P<0.05).Conclusion:Xiaochaihu decoction can inhibit the activity of HBV in HepG2.2.15 cells and protect it,and there is a possibility to regulate the key signal transduction pathway(JAK-STAT)to influence the development of CHB.