Effect Of Paeonol On Expression And Function Of Drug Transporters In Human Hepatoma Cell Lines HepG2

Paeonol(Pae),which has wide pharmacological activity,is one of the main active component of plants of root bark of Ranunculaceae peony Paeonia suffrutlcosa Andr and dried root and rhizome of Asclepiadaceae plants Cynanchum paniculatum (Bge.) Kitag. According to the related literatures, it has certain effect in anti-inflammatory, antiallergic, antipyretic analgesic, antihypertensive, anti-atherosclerosis and anti-tumour. The reverse MDR effect of Paeonol in anti-tumor aspect, finally make the anticancer drug to maintain a high concentration in cells. The process of drug transport in vivo involve the special transport protein system, which called transporters. In our research, the effects of paeonol on expression and function of drug transporters in human hepatoma cell lines HepG2 and the possible mechanism will be investigated.1. Effect of Pae on the proliferation of human hepatoma cell lines HepG2CCK8 was used to detect the dose and time dependence of pae on the proliferation of HepG2.We found that the concentration of pae(2～8 μg.mL-1) has no significantly inhibition on HepG2,except the concentration of pae(16 μg.mL-1);and the inhibitory effects is no less 10% after pae (8 μg.mL-1) culture time for 6、12、2、48 h.Combined with the experiment in vivo,we will used pae(2μg.mL-1)culture HepG2 for 24 h.2.Effect of pae on expression and function of human hepatoma cell lines HepG2qRT-PCR and flow cytometry were used to detect the mRNA and activity of transporters. We found that the mRNA of OATP1B1、OATP1B3、 OATP2B1、BCRP and MRP2 were induced,except OAT2 and MRP4.Moreover, activity of OATP1B1 and BCRP was up-regulated by pae.The results suggest that pae may induced activity of OATP1B1、 OATP1B3、OATP2B1、BCRP and MRP2 through up-regulating the mRNA,except OAT2 and MRP4.3.the mechanism study of the effect of pae on up-regulation the expression of some transporters in human hepatoma cell lines HepG2qRT-PCR was performed to measure the expression of PXR mRNA expression.After culture by paeonol(2μg.mL-1) for 24h,the expression of PXR mRNA was up-ragulated,and it implicates that the expressin of transporters is with the relation of the nuclear receptor PXR pathway. In order to verify the above conclusions,PXR inhibitor ketoconazole was used to culture cells for 24 h with paeonol.Compared with Pae(2μg.mL-1) culture group,the expression of MRP2 at the level of transcription, protein levels were down-regulated,activity also inhibited by qRT-PCR,WB and flow cytometry.In summary, paeonol may regulate the expression and function of some transporters through up-regulation the level of PXR in HepG2 cells.