Study of the effects of Berberine on CYP3A4 and P-gp in HepG2 cells and its mechanism in vitro

Abstract

Abstract: AIM: To study the effects of berberine hydrochloride(BBR) on the mRNA and protein expression of cytochrome P450 3A4(CYP3A4) and multidrug resistance 1(MDR1) in HepG2 cells, then to investigate the mechanisms of regulation of CYP3A4 and P-glycoprotein(P-gp) by berberine. METHODS: HepG2 cells were treated with different concentrations of BBR(1-100 μg/mL)for 48 h, then effects of BBR on the cell proliferation were detected by MTT assay. The experiments were taken when the cytotoxicity of BBR on HepG2 cells was low.The experiments were divided into the following groups: control group, groups administrated with different concentrations of BBR(0.1-40 μg/mL), 10 μmol/L Rifampicin(Rif)group and groups administrated with different concentrations of BBR (0.1-40 μg/mL) and together with 10 μmol/L Rif, incubated with the logarithmic phase of HepG2 cells for 48 h. Total proteins and RNA were extracted. The mRNA levels of CYP3A4, MDR1, pregnane X receptor (PXR) and retinoid X receptor (RXR) in HepG2 cells were assayed with QRT-PCR, and the protein levels of CYP3A4,P-gp in HepG2 cells were assayed with Western blot. RESULTS: When the concentration of BBR was during 1- 40 μg/mL, the cytotoxicity of BBR on HepG2 cells was low, besides, the living rates of HepG2 cells were all above 80%.Compared with the control group,Western blot analysis showed that groups administrated with 0.1,0.5 and 2 μg/mL BBR significantly induced the expression of CYP3A4 and P-gp proteins in HepG2 cells(P<0.05). However, the 10 and 40 μg/mL BBR groups could markedly inhibit the expression of CYP3A4 and P-gp proteins in HepG2 cells(P<0.01).QRT-PCR analysis indicated that groups administrated with 10, 20 and 40 μg/mL BBR could strongly down-regulated the CYP3A4, PXR, and MDR1 mRNA expression in HepG2 cells(P<0.05) but 10 μg/mL BBR group had no significant inhibitory effects on the RXR mRNA expression(P>0.05),20 and 40 μg/mL BBR groups markedly inhibited the RXR mRNA expression(P<0.05). CONCLUSION: Berberine had a dual role on the expression of CYP3A4 and P-gp. Induction was leaded by low doses berberine, inhibition was leaded by high doses berberine. The mechanisms of regulation of CYP3A4 and P-gp by berberine was probably through PXR pathways.

Key words: berberine hydrochloride, CYP3A4, P-gp, PXR, RXR, drug interactions

CLC Number:

R965.2

TANG Xia, XIN Hua-wen, LI Wei-liang, OUYANG Meng. Study of the effects of Berberine on CYP3A4 and P-gp in HepG2 cells and its mechanism in vitro[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2015, 20(1): 7-13.

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