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PXR-Mediated Upregulation of CYP3A Expression by Herb Compound Praeruptorin C from Peucedanum praeruptorum Dunn.

Huang L, Wu Q, Li YH, Wang YT, Bi HC - Evid Based Complement Alternat Med (2013)

Bottom Line: We recently reported that Praeruptorin C effectively transactivated the mRNA, protein expression, and catalytic activity of CYP3A4 via the CAR-mediated pathway, but whether and how PC could affect the expression and catalytic activity of CYP3A4 via PXR pathway remains unknown.In PXR-overexpressed LS174T cells, PC significantly enhanced CYP3A4 mRNA, protein expression, and functional activity through PXR-mediated pathway; conversely, no such increase was found in the untransfected cells.These findings suggest that PC can significantly upregulate CYP3A level via the PXR-mediated pathway, and this should be taken into consideration to predict any potential herb-drug interactions between PC, Qianhu, and the other coadministered drugs.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan Dong Road, Guangzhou 510006, China ; School of Pharmaceutical Sciences, Hainan Medical University, Haikou 571010, China.

ABSTRACT
We recently reported that Praeruptorin C effectively transactivated the mRNA, protein expression, and catalytic activity of CYP3A4 via the CAR-mediated pathway, but whether and how PC could affect the expression and catalytic activity of CYP3A4 via PXR pathway remains unknown. Therefore, in this study, the effect of PC on the CYP3A gene expression was investigated in mice primary hepatocytes after knockdown of PXR by transient transfection of PXR siRNA, and the gene expression, protein expression, and catalytic activity of CYP3A4 in the LS174T cells with PXR overexpression were determined by real-time PCR, western blot analysis, and LC-MS/MS-based CYP3A4 substrate assay, respectively. We found that the level of CYP3a11 gene expression in mouse primary hepatocytes was significantly increased by praeruptorin C, but such an induction was suppressed after knockdown of pregnane X receptor by its siRNA. In PXR-overexpressed LS174T cells, PC significantly enhanced CYP3A4 mRNA, protein expression, and functional activity through PXR-mediated pathway; conversely, no such increase was found in the untransfected cells. These findings suggest that PC can significantly upregulate CYP3A level via the PXR-mediated pathway, and this should be taken into consideration to predict any potential herb-drug interactions between PC, Qianhu, and the other coadministered drugs.

No MeSH data available.


PC physicochemical properties investigated in this study. CAS Number: 73069-27-9. Molecular Formula: C22H22O8.
© Copyright Policy - open-access
Related In: Results  -  Collection


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fig1: PC physicochemical properties investigated in this study. CAS Number: 73069-27-9. Molecular Formula: C22H22O8.

Mentions: Praeruptorin C (Figure 1) (purity > 99%) was available from Kui Qing Chemical Company (Tianjin, China). Dimethyl sulfoxide (DMSO), dexamethasone (DEX), nifedipine (NIF), dehydronifedipine (DNIF), and loratadine were purchased from Sigma-Aldrich (St. Louis, MO, USA). The validated siRNA targeted to the PXR gene and nontargeting siRNA as a silencer negative control were purchased from Guangzhou RiboBio Co., Ltd (Guangzhou, China). siRNA Transfection Reagent was purchased from Roche (New Jersey, USA). RNAiso Plus and PrimeScriptTM RT Reagent were obtained from Takara (Kyoto, Japan). The Cyp3a11, PXR, CYP3A4, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) primers used in real-time PCR were synthesized by Takara. Anti-CYP3A4 polyclonal antibody was purchased from Millipore Corporation (Rosemont, IL, USA); anti-Cyp3a11 polyclonal antibody and GADPH antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-rabbit IG-HRP antibody was purchased from R&D Systems (Minneapolis, MN, USA). SDS-PAGE Gel Preparation Kit was purchased from Beyotime Institute of Biotechnology (Haimen, Jiangsu, China). Plasmocin TM Ant-mpp and complete protease inhibitor cocktail were purchased from Invitrogen (San Diego, CA, USA) and Roche Diagnostics (Basel, Switzerland), respectively. The cytotoxicity of LS174T cells were detected by the 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) cytotoxicity assay, and PA did not show cytotoxicity in LS174T cells under the maximum dosage (40 μM).


PXR-Mediated Upregulation of CYP3A Expression by Herb Compound Praeruptorin C from Peucedanum praeruptorum Dunn.

Huang L, Wu Q, Li YH, Wang YT, Bi HC - Evid Based Complement Alternat Med (2013)

PC physicochemical properties investigated in this study. CAS Number: 73069-27-9. Molecular Formula: C22H22O8.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3860132&req=5

fig1: PC physicochemical properties investigated in this study. CAS Number: 73069-27-9. Molecular Formula: C22H22O8.
Mentions: Praeruptorin C (Figure 1) (purity > 99%) was available from Kui Qing Chemical Company (Tianjin, China). Dimethyl sulfoxide (DMSO), dexamethasone (DEX), nifedipine (NIF), dehydronifedipine (DNIF), and loratadine were purchased from Sigma-Aldrich (St. Louis, MO, USA). The validated siRNA targeted to the PXR gene and nontargeting siRNA as a silencer negative control were purchased from Guangzhou RiboBio Co., Ltd (Guangzhou, China). siRNA Transfection Reagent was purchased from Roche (New Jersey, USA). RNAiso Plus and PrimeScriptTM RT Reagent were obtained from Takara (Kyoto, Japan). The Cyp3a11, PXR, CYP3A4, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) primers used in real-time PCR were synthesized by Takara. Anti-CYP3A4 polyclonal antibody was purchased from Millipore Corporation (Rosemont, IL, USA); anti-Cyp3a11 polyclonal antibody and GADPH antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-rabbit IG-HRP antibody was purchased from R&D Systems (Minneapolis, MN, USA). SDS-PAGE Gel Preparation Kit was purchased from Beyotime Institute of Biotechnology (Haimen, Jiangsu, China). Plasmocin TM Ant-mpp and complete protease inhibitor cocktail were purchased from Invitrogen (San Diego, CA, USA) and Roche Diagnostics (Basel, Switzerland), respectively. The cytotoxicity of LS174T cells were detected by the 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) cytotoxicity assay, and PA did not show cytotoxicity in LS174T cells under the maximum dosage (40 μM).

Bottom Line: We recently reported that Praeruptorin C effectively transactivated the mRNA, protein expression, and catalytic activity of CYP3A4 via the CAR-mediated pathway, but whether and how PC could affect the expression and catalytic activity of CYP3A4 via PXR pathway remains unknown.In PXR-overexpressed LS174T cells, PC significantly enhanced CYP3A4 mRNA, protein expression, and functional activity through PXR-mediated pathway; conversely, no such increase was found in the untransfected cells.These findings suggest that PC can significantly upregulate CYP3A level via the PXR-mediated pathway, and this should be taken into consideration to predict any potential herb-drug interactions between PC, Qianhu, and the other coadministered drugs.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan Dong Road, Guangzhou 510006, China ; School of Pharmaceutical Sciences, Hainan Medical University, Haikou 571010, China.

ABSTRACT
We recently reported that Praeruptorin C effectively transactivated the mRNA, protein expression, and catalytic activity of CYP3A4 via the CAR-mediated pathway, but whether and how PC could affect the expression and catalytic activity of CYP3A4 via PXR pathway remains unknown. Therefore, in this study, the effect of PC on the CYP3A gene expression was investigated in mice primary hepatocytes after knockdown of PXR by transient transfection of PXR siRNA, and the gene expression, protein expression, and catalytic activity of CYP3A4 in the LS174T cells with PXR overexpression were determined by real-time PCR, western blot analysis, and LC-MS/MS-based CYP3A4 substrate assay, respectively. We found that the level of CYP3a11 gene expression in mouse primary hepatocytes was significantly increased by praeruptorin C, but such an induction was suppressed after knockdown of pregnane X receptor by its siRNA. In PXR-overexpressed LS174T cells, PC significantly enhanced CYP3A4 mRNA, protein expression, and functional activity through PXR-mediated pathway; conversely, no such increase was found in the untransfected cells. These findings suggest that PC can significantly upregulate CYP3A level via the PXR-mediated pathway, and this should be taken into consideration to predict any potential herb-drug interactions between PC, Qianhu, and the other coadministered drugs.

No MeSH data available.