The In Vitro Study About Jianpi Qinghua Decoction Drug Serum On HepG2 Cell Infected With HCV

Objective:1.Establishing the Hep G2 cell model infected with postive serum of hepatitis C with hepatic steatosis.2.Observing the influence of Jianpi Qinghua Decoction drug serum to apolipoprotein B(Apo B),very low density lipoprotein(VLDL),triglyceride(TG),the quantity of HCV-RNA in the hepatic cellular cancer cell superntant,and the effect to the expression of MTP,Apo B protein in the Hep G2 cell.3.To illuminate the curative effect of Jianpi Qinghua Decoction to the interferon against hepatitis C virus and the mechanism that it did.Methords:1.Dividing the forty SPF SD male rats with the method of randomized into such groups as western medicine group,chinese medicine group,combined group and control group,10 rats of each group,accordingly were given peginterferon subcutaneous injection plus ribavirin lavage,Jianpi Qinghua Decoction lavage,peginterferon subcutaneous injection plus ribavirin and Jianpi Qinghua Decoction lavage,the control group was given saline gavage,10 days later,produced four groups of rats' abdominal cavity serum.2.Establishing the Hep G2 cell model infected with postive serum of hepatitis C with hepatic steatosis,and use reverse transcription polymerase chain reaction(RT-PCR)to detect HCV-RNA positive strand.If it is positive,suggesting the model is successful.3.Useing rats drug serum of the each group to respectively intervene the Hep G2 cell model.After that cultured the cell for forty-eight hours(two days),then collecting the cell and cell superntant.4.Detecting the variation of content of Apo B,VLDL,TG in the in the Hep G2 cell superntant by ELISA,the quantity of HCV-RNA by RT-PCR,using Western blot method to detect Apo B,MTP protein expression of the cell of eachgroup.Results:The group before model establishment namely before infected with HCV was assumed as blank group,after model establishment namely before infected with HCV was model group.1.The content of Apo B,VLDL in the in the Hep G2 cell superntant after infected with HCV,namely model group,declined compared with blank group,and the content of TG rised.2.The content of Apo B,VLDL,in the Hep G2 cell superntant after intervened by the rats drug serum of the each group went up in different degree compared with control group,the content of TG declined.And the most obvious change(rised/declined)is the peginterferon plus ribavirin and Jianpi Qinghua Decoction group,namely combined group,next is Jianpi Qinghua Decoction group,namely chinese medicine group,peginterferon plus ribavirin group,namely western medicine group is the last one,which suggesting that combined group had the greatest effect on lipid metabolism,then is the chinese medicine group,the western medicine group had no obvious effect(P>0.05).3.The quantity of HCV-RNA in the in the Hep G2 cell superntant intervened by the rats drug serum of the each group declined compared with control group.And the most obvious change is the combined group,next is the western medicine group,the chinese medicine group is the last one,which prompting that combined group had the greatest effect on the change of quantity of HCV-RNA,the next is the western medicine group,it has no obvious influence for the chinese medicine group(P>0.05).4.After infected with HCV,namely model group,in the Hep G2 cell,the relative transcript level of MTP,Apo B protein decreased compared with blank group.But after the rats drug serum interven Hep G2 cell,the relative transcript level of MTP,Apo B protein increased in different degree compared with control group,And the most obvious change is the combined group,it had no significant difference between the western medicine group and the chinese medicine group(P > 0.05),which suggesting that combined group had the greatest effect on the expression of MTP,Apo B protein.Conclusion:1.The content of Apo B,VLDL in the Hep G2 cell superntant after infected with HCV,namely model establishment sucessfully,dropped compared with that before model establishment,and the conten of TG rised.The relative transcript level of MTP,Apo B protein also dropped after model establishment sucessfully.2.Jianpi Qinghua Decoction can improve the content of Apo B,VLDL effectively,reduce the TG,accelerate the expression of Apo B,MTP protein,and then enhance that interferon inhibit HCV-RNA replication.3.The mechanism of Jianpi Qinghua Decoction can enhance the efficacy of interferon against HCV may be that it can regulate the MTP-Apo B lipid transport pathway,and promote the MTP combine with Apo B which riched in TG in the endoplasmic reticulum lumen,and carried out liver in the form of VLDL.