The Effect Of Emodin On The Signaling Pathway Of FXR-BSEP In The Human Hepatocyte LO2 Cell Line

Objectives: To investigate the effect of Emodin on the expression of FXR and its downstream BSEP gene in human hepatocyte LO2 cell line.Methods: Human hepatocyte LO2 cells were seeded in DMEM high glucose medium supplemented with 13% fetal bovine serum.The m RNA and protein levels of FXR and BSEP were detected respectively by Real-Time PCR and Western-blot technology at low,medium,high dose(0.02,0.04,0.08μg/ml)of Emodin,with UDCA(0.1μg/ml),dexamethasone(0.785ng/μl)as positive control.With the inhibitor Guggulsterones of FXR down regulating its expression,examine the m RNA and protein levels of FXR and BSEP by Real-Time PCR and Western-blot technology at low,medium,high dose of Emodin,with UDCA,dexamethasone as positive control.Up regulating the expression of FXR with the agonist GW4064,the m RNA and protein levels of FXR and BSEP were assayed by Real-Time PCR and Western-blot technology at low,medium,high dose of emodin,with UDCA,dexamethasone as positive control.Results: 1.Real-Time PCR: Eomdin can stimulate m RNA expression of FXR and BSEP compared with the control group(P<0.05).2.Western-blot: In the case of the normal expression of FXR,the groups with three doses of Emodin were significantly higher than the normal control group in the expression of FXR protein.When down-regulating FXR expression with the inhibitor Guggulsterones,FXR protein expression in the groups with medium,high doses of Emodin were higher than that of the Guggulsterones control group(P<0.01),while the three doses of Emodin can stimulate more BSEP protein than Guggulsterones(P<0.01).When up-regulating FXR expression with the agonist GW4064,the expression of FXR protein in the group with high dose of Emodin was higher than that of GW4064 control group(P<0.01).Conclusion: Emodin can,to a certain degree,stimulate the expression of FXR and its downstream BSEP gene in the human hepatocyte LO2 cell line at the cellular level.However,whether there are other signaling pathways effecting the two genes and whether Emodin can really play a role in promoting the transportation of accumulated bile remains to be further studied.