The Regulation Mechanism Of Tea Polyphenols On CYP450s In C57BL/6J Mouse And HepG2 Human Liver Cancer Cells

In this study,we used three different fermentation stages teas(Long Jing,Tie Guan Yin and Dian Hong)as raw materials,the tea polyphenol were extracted by methanol.The obtained tea polyphenol extracts were given to C57BL/6J mouses 7 days respectively,Real-time reverse transcription polymerase chain reaction(real-time PCR)and Western blot were used to investigate the dose-effect relationship between the mRNA and protein expression of Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37,Cyp1a2 in mouse liver Cyp450s;C57BL/6J mouses were inoculated with the three kinds of tea polyphenol extracts at the appropriate dose respectively 7,14,28 days,and the time-effect relationship between the mRNA and protein expression of mouse liver Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37,Cyp1a2 in mouse liver were investigated.The quantitative analysis of the major catechins in the three kinds of tea polyphenol extracts was performed by HPLC.The expression of CYP3A4,CYP2E1,CYP2D6,CYP2C9,CYP1A2 mRNA and protein was detected by human liver cancer Hep G2 with the main catechins and tea polyphenol extracts.To provide a theoretical basis for the mechanism of tea polyphenol extracts on the regulation of drug metabolizing enzyme of liver.The main conclusions are as follows:(1)The dose-effect relationship of Long Jing,Tie Guan Yin and Dian Hong polyphenol extracts on the expression of Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37 and Cyp1a2 in mouse liver Cyp450 s were investigated.The results showed that: The Long Jing polyphenol extracts on the expression of Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37,Cyp2d22 mRNA and protein have a certain dose-effect relationship,there is a downward trend and a significant impact(p<0.05).The Tie Guan Yin polyphenol extracts could significant down-regulated the expression of on Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37,Cyp1a2 mRNA and the protein of Cyp2e1,Cyp2d22,Cyp2c37 and Cyp1a2(p<0.05).The Dian Hong polyphenol extracts could significant affected the mRNA and protein expression of on Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37 and Cyp1a2(p<0.05).(2)The time-effect relationship of Long Jing,Tie Guan Yin and Dian Hong polyphenol extracts on mouse liver Cyp450 s was discussed.The results showed that: Long Jing polyphenol extracts inhibited significantly the mRNA expression of Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37,Cyp1a2 in mice after gavaging 7,14,28 days(p<0.05);they has a significant effect on the protein expression of Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37,Cyp1a2(p<0.05).The mRNA expression of Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37,Cyp1a2 was significantly induced by Tie Guan Yin polyphenol extracts after 7,14,28 days(p<0.05),and has a significant effect on these protein expression(p<0.05).The expression of mRNA and protein of Cyp3a11,Cyp2e1,Cyp2d22,Cyp2c37 and Cyp1a2 was significant at 7,14 and 28 days after intragastric administration by Dian Hong polyphenol extracts(p<0.05).(3)The components and contents of Long Jing,Tie Guan Yin,Dian Hong polyphenol extracts were detected by HPLC.The results showed that: in Long Jing polyphenol extracts,they were mainly containing epigallocatechin gallate(EGCG),epigallia(EGC),epicatechingallate(ECG),respectively 288.83,57.74,42.08 ?g/mg;Tie Guan Yin polyphenol ertracts were with the higher content of EGC,EGC and epicatechin(EC),129.21,71.49,17.78 ?g/mg,respectively,and the ECG was 16.78 ?g/mg.Dian Hong polyphenol ertracts has EGCG,ECG,gallic acid(GA),34.70,30.42,24.48 ?g/mg respectively,among them,Epigallocatechin(EGC)was 6.75 ?g/mg.(4)The mRNA and protein expression of CYP450 s in Hep G2 cells were studied by Long Jing,Tie Gguan Yin and Dian Dong polyphenol extracts.The results showed that: the mRNA expression of CYP1A2,CYP2C9 and CYP2E1 were significantly inhibited by Long Jing polyphenol extracts(p<0.05),but the mRNA expression of CYP3A4 and CYP2D6 was significantly induced(p<0.05).The protein expression of CYP3A4,CYP2E1,CYP2D6,CYP2C9 and CYP1A2 were significantly inhibited by Long Jing polyphenol extracts(p<0.05).the mRNA expression of CYP1A2,CYP2C9 and CYP2E1 were significantly inhibited and he mRNA expression of CYP3A4(p<0.05),CYP2D6 were significantly induced by Tie Guan Yin polyphenol extracts(p<0.05).the protein expression of CYP3A4,CYP2D6,CYP1A2,CYP2C9 and CYP2E1 was significantly inhibited.the mRNA expression of CYP2E1,CYP2D6,CYP2C9 and CYP1A2 was significantly indused and the protein expression of CYP2A1,CYP2D1,CYP2D6,CYP1A2 and CYP2C9 was significantly inhibited by the Dian Hong polyphenol extracts(p<0.05).(5)The mRNA and protein expression of CYP450 s in Hep G2 was studied by EGCG,EGC and ECG,which were the main concents.The results showed: EGCG,EGC and ECG significantly inhibited the mRNA expression of CYP1A2,CYP2C9 and CYP2E1,respectively(p<0.05),but the mRNA expression of CYP3A4 and CYP2D6 were significantly induced(p < 0.05).EGCG significantly inhibited the protein expression of CYP3A4(p<0.05),EGC and ECG significantly induced the protein expression of CYP3A4(p<0.05).EGCG significantly inhibited the protein expression of CYP2E1,CYP2D6,CYP2C9 and CYP1A2(p<0.05).EGC significantly inhibited the protein expression of CYP2D6 and CYP2E1(p<0.05),but significantly induced the protein expression of CYP1A2.ECG had a significant effect on the protein expression of CYP1A2,CYP2E1,CYP2C9 and CYP2D6(p<0.05).