The Regulation Of Longbishu On MRNA And Protein Expression Of Cytochrome P450 Isoenzymes In Rat Liver.

Objective: Longbishu capsule is a kind of compound Chinese medicine preparation used to treat benign prostatic hyperplasia and chronic prostatitis in clinical. Long-term administration of longbishu may affect the cytochrome P450 isoenzymes, and further cause drug-drug interactions. In this study, we investigated changes of m RNA and protein expression of cytochrome P450 isoenzymes in rat liver after intragastric administration of longbishu for two months.Methods: Sixteen healthy male SD rats, weighting 180-200 g, were randomly divided into solvent control(CON) group and longbishu(LBS) group. The solvent control group was intragastrically administered by 0.5% sodium carboxymethyl cellulose(CMC-Na) solution, and the longbishu group was intragastrically administered by longbishu powder at the dosage of 154.3mg/kg(5 times higher than the therapeutic dosage in clinical). The longbishu suspension was preparaed by 0.5% CMC-Na solution. After intragastric administration of longbishu for two months, the rats were killed on the next day after the last administration of drug. The liver was perfused with normal saline, and turned into muddy color by in situ brain perfusion technique, then quickly excised about 100 mg kept into D/RNase-free cryopreservation tubes and another 2g was used to prepare the liver microsomals. Samples were stored at-80 ℃ for later use. The m RNA expression levels of CYP1A2, CYP2B1, CYP2C11, CYP2D1, CYP2E1, CYP3A1 and CYP3A2 subtypes were investigated by Real-time PCR technique and the protein expression levels of CYP1A1, CYP1A2, CYP2C11, CYP2D1, CYP2E1 and CYP3A1 subtypes were observed by Western-blot technique.1 The effects of longbishu on m RNA expression of CYP subtypes in rat liver.Compared with the control group, the m RNA expression levels of CYP1A2, CYP2B1, CYP2C11 CYP2E1, CYP3A1, and CYP3A2 was increased significantly in rat liver after intragastric administration of longbishu for two months(P<0.01). Set the m RNA expression levels of control group as a benchmark to 1. The m RNA relative expression levels of CYP1A2, CYP2B1, CYP2C11, CYP2E1, CYP3A1 and CYP3A2 were 2.20±0.16, 1.85±0.16, 1.29±0.05, 1.52±0.09, 1.44±0.06 and 1.78±0.10 in rat liver of the longbishu group.Compared with the control group, the m RNA expression level of CYP2D1 did not significantly change in rat liver(P>0.05). The m RNA relative expression of CYP2D1 was 1.06±0.04 in rat liver of the longbishu group.2 The effects of longbishu on protein expression of CYP subtypes in rat liver.Compared with the control group, the protein expression levels of CYP1A1, CYP1A2, CYP2C11, CYP2D1, CYP3A1 was significantly increased in rat liver after intragastric administration of longbishu for two months(P<0.01). The ratios of CYP1A1, CYP1A2, CYP2C11, CYP2D1, CYP3A1 to GAPDH were 0.39±0.02, 0.63±0.02, 1.32±0.03, 1.88±0.04, 0.61±0.02 in the longbishu group and 0.32±0.02, 0.41±0.01, 1.08±0.05, 1.56±0.02, 0.44±0.01 in the control group.Compared with the control group, the protein expression level of CYP2E1 was decreased significantly in rat liver(P<0.05). The ratios of CYP2E1 to GAPDH were 0.19±0.01 and 0.24±0.01, respectively in longbishu and control groups.Conclusion:1 The m RNA expression levels of CYP1A2, CYP2B1, CYP2C11, CYP2E1, CYP3A1 and CYP3A2 were increased significantly in rat liver after intragastric administration of longbishu for two months.Results:2 The protein expression level of CYP2E1 was decreased significantly and the protein expression levels of CYP1A1, CYP1A2, CYP2C11, CYP2D1 and CYP3A1 were increased significantly in rat liver after intragastric administration of longbishu for two months.