Expression Of P-gp/MDR1 And ABCG2/BCRP In Placenta With Hepatitis B Virus Infection

BackgroundP-gp/MDR1 and ABCG2/BCRP are members of the group of transporters in the ATP binding cassette(ABC) superfamily,which are localized in the syncytiotrophoblast of the human placenta.These proteins actively extrudes a wide variety of drugs and toxins from cells,including antiretroviral HBV drugs,such as zidovudine and lamivudine.Regulation of P-gp/MDR1 and ABCG2/BCRP expression may improve access of the antiretroviral drugs to the fetus,so as to prevent materal-fetal transmission of HBV during pregnance.ObjectiveTo determine the expression of P-gp/MDR1 and ABCG2/BCRP in full-term placentas with HBV infection,and evaluate the relation between pro-inflammatory cytokines with P-gp/MDR1 and ABCG2/BCRP expression.Study design1.P-gp/MDR1 and ABCG2/BCRP in full-term placentas were localized by immunohistochemistry.2.P-gp/MDR1mRNA and ABCG2/BCRP mRNA transcripts were quantified by real-time PCR using relative determinations.The levels of P-gp/MDR1 mRNA and ABCG2/BCRP mRNA in placentas with HBV infection were compared with those of the uninfected groups.3.The profile of P-gp/MDR1 and ABCG2/BCRP protein levels were quantified using Western blot analysis.P-gp/MDR1 and ABCG2/BCRP protein levels in placentas with HBV infection were compared with those of the uninfected groups.4.Concentrations of IL-1β,IL-6 and TNF-αproteins in placentas were determined using Enzyme-linked immunosorbent assay(ELISA).The relation between pro-inflammatory cytokines with P-gp/MDR1 and ABCG2/BCRP expression were evaluated.5.BeWo and JEG-3 cells were treated with IL-1βor IL-6 for different time periods(12 h,24 h,48 h,72h),then the P-gp/MDR1mRNA and ABCG2/BCRP mRNA transcripts were quantified by real-time PCR using relative determinations.6.The concentrations of lamivudine acrossed the cells were determined using HPLC in BeWo cells treated with IL-1βor IL-6.Results1.Immunohistochemically P-gp/MDR1 and ABCG2/BCRP proteins were localized on the maternal side membrance of syncytiotrophoblast in full-term placentas with or without HBV infection.2.The levels of P-gp/MDR1mRNA and ABCG2/BCRP mRNA in placentas with HBV infection were statistically significantly higher than those of the, non-infected groups using real-time quantitative PCR.3.The Western blot analysis showed a statistically significant increase in P-gp/MDR1 and ABCG2/BCRP protein levels in placentas with HBV infection compared with those of the non-infected groups.P-gp/MDR1 and ABCG2/BCRP protein levels in placentas with HBV infection were 2.43 and 3.76 times higher than those of the non-infected groups.4.Concentrations of IL-1β,IL-6 and TNF-αproteins in placentas with HBV infection were statistically significantly higher than those of the non-infected groups determined by Enzyme-linked immunosorbent assay.Concentrations of IL-1β,IL-6 and TNF-αin placentas with HBV infection were 4.36,1.91,5.12 times higher than those of the non-infected groups.And the increase of IL-1β, IL-6 and TNF-αwere related to the expression of P-gp/MDR1 and ABCG2/BCRP.5.Relative quantitative PCR showed a statistically significant increase in P-gp/MDR1 mRNA and ABCG2/BCRP mRNA transcription in BeWo and JEG-3 cells treated with IL-1βor IL-6 at 12h,24h,48h and 72h later.And the transcription level was increased remarkably with correspondence to time.6.The concentrations of lamivudine acrossed the cells were significantly decreased in BeWo cells treated with IL-1βor IL-6 determined by HPLC.Conclusion1.P-gp/MDR1 and ABCG2/BCRP proteins express in full-term placentas of both HBV infected and non-infected groups,and localized to the maternal side membrance of syncytiotrophoblast.2.The levels of P-gp/MDR1 mRNA and ABCG2/BCRP mRNA in placentas with HBV infection are significantly higher than those of the non-infected groups.3.The P-gp/MDR1 and ABCG2/BCRP protein levels in placentas with HBV infection are significantly higher compared with those of the non-infected groups.4.Concentrations of IL-1β,IL-6 and TNF-αproteins in placentas with HBV infection are significantly higher than those of the non-infected groups.And the increase of IL-1β,IL-6 and TNF-αare related to the expression of P-gp/MDR1 and ABCG2/BCRP.5.P-gp/MDR1 mRNA and ABCG2/BCRP mRNA transcription are significantly increased in BeWo and JEG-3 cells after treated with IL-1βor IL-6.IL-1βand IL-6 may activate the transcription of P-gp/MDR1 mRNA and ABCG2/BCRP mRNA.6.The infiltration rates of lamivudine acrossed the cells are significantly decreased in BeWo cells after treated with IL-1βor IL-6,which maybe related to the up regulation of P-gp/MDR1 mRNA and ABCG2/BCRP mRNA.