| PurposeTo study the induction of Composite Salvla Recipe affects hepatic and cardiaccytochrome P450expression main subtypes in rats; To study the induction of Panaxnotoginseng saponin（PNS） affects cytochrome P450mRNA expression in H9c2cell;In order to understand the preventive and therapeutic role of artemisinin oninflammatory bowel disease (IBD) and whether Pregnane X Receptor (PXR) activationis dependent human colon cancer-derived LS174T cells were used to investigate theability of artemisinin to activate the PXR signaling pathway and to induce theexpression of CYP3A4.Method1. Male SD rats were divided into six groups and rat liver microsomes were taken aftera28-day gavage (ig). A HPLC-MS method was applied to determine the enzymeactivity of CYP1A2,2B6,2C12,2C13,2D2and3A1.2. The expression of cyp1a2,2b1/2,2c11,2e1and3a1mRNA were determined byRT-PCR.3. Male SD rats were randomly divided into five groups and cardiac tissues were takenafter a28-day stomach tube (ig). The expression of cardiac cytochrome P450mRNAswere detected by Real Time PCR. 4. H9c2cells were treated with different concentrations of PNS for48h.Cell viabilitywas examined by MTS assay and cell menbrane injury was examined by detecting therelease rate of lactate dehvdrogenase (LDH). The expression of cytochrome P450mRNAs were detected by Real Time PCR.5. CYP3A4and PXR mRNA expression and protein expression were detected by RealTime PCR and Western-blot respectively on cell level. The dextran sulfate sodium(DSS)-induced IBD model was constructed and treated with artemisinin, in order tostudy that artemisinin and PXR might merit attention in the treatment of IBD.Results1. Compared with the control group, phenobarbital group had inhibitory effects on theenzyme activity of CYP2D2and3A1, while the activities of CYP1A1,2B6,2C12and2C13were induced (P＜0.05or P＜0.01); Composite Salvla Recipe had inhibitoryactivity of CYP1A2and2B6, while induced CYP2D2(P＜0.05); Danshen inhibitedthe activities of CYP1A2and2B6(P＜0.05); Panax notoginseng inhibited CYP1A2,2B6,2C13and2D2(P＜0.05or P＜0.01); Borneol had inhibitory effects onCYP1A2,2B6,2C12,2C13and2D2(P＜0.01). On the mRNA level, compared withthe control group, benzene barbitone group had inductive effects on the liver mRNAof cyp1a2and2b1/2(P＜0.05); Composite Salvla Recipe, Danshen and panaxnotoginseng had no significant effects on the mRNA level of cyp1a2,2b1/2,2c11,2e1and3a1; Borneol significantly inhibited the mRNA Level of cyp1a2,2b1/2and2c11(P＜0.05or P＜0.01).2. Compared with the control group, Composite Salvla Recipe group had inhibitioneffects on the mRNA level of CYP1B1, CYP2B1, CYP2E1, CYP4A1and CYP4F4(P＜0.05). Danshen group significantly inhibited the mRNA level of CYP1A1,CYP1B1, CYP2B1, CYP2C11, CYP2E1, CYP2J3, CYP4A1, CYP4F4and CYP4F5 (P＜0.05or P＜0.01), while induced the mRNA level of CYP4A3, CYP4F1andCYP4F6(P＜0.05). Panax notoginseng group had inhibition influence on themRNA level of CYP1A1, CYP1B1, CYP2B1, CYP2C11, CYP2E1, CYP2J3,CYP4A1, CYP4A3, CYP4F4, CYP4F5and CYP4F6(P＜0.05or P＜0.01).Borneol group inhibited the mRNA level of CYP1A1, CYP1B1, CYP2B1, CYP2C11,CYP4A1and CYP4F4(P＜0.01).3. H9c2cell viability was significantly reduced following exposure to dictamnine0.3125～20mg.ml-1for48h in a concentration-dependent manner and IC50value was（5.28±0.08）mg·ml-1.The LDH release rate of H9c2cells was significantlyincreased after exposure to dictamnine0.3125～20mg·ml-1for48h (P＜0.01). Onthe mRNA level, compared with the control group, PNS induced the mRNA level ofCYP2J3, CYP4A1, CYP4A2, CYP4F1and CYP4F5(P＜0.05or P＜0.01). PNShad inhibitory activity of CYP2C11, CYP4A3and CYP4F4(P＜0.05or P＜0.01), while had inductive effects mRNA of CYP4A3and CYP4F4(P＜0.01) at5mg.ml-1.4. Results showed CYP3A4and PXR mRNA and protein levels to be increased byartemisinin in a dose-dependent manner. Artemisinin was mediated throughactivation of PXR and initiated by PXR-PXRE binding. A DSS-induced IBD modelwas constructed, and treatment with artemisinin decreased the severity of colitis asindicated by mouse body weight, colon length, diarrhea, and rectal bleeding and byboth macroscopic and histological analysis. The expression of mRNAs encodingCyp3a11, Cyp3a13, GSTα1, and MDR1α were higher in vehicle-, artemisinin-, andPCN-treated mice than in DSS-treated mice. The results suggest that PXR-mediatedsuppression of NF-κB target genes (CCR2, ICAM-1, IL-1β, IL-6, IL-10, iNOS,MCP-1, and TNFа) may be the mechanism by which PXR provides protection toDSS-induced IBD. Conclusion1. These results indicated that single drug had more significant influence on enzymesthan Composite Salvla Recipe. Borneol had the most significant inhibitory effect ondrug metabolic enzymes,. And the interactions between drugs were minimized. Thisis beneficial for safe medication, reflecting the whole party formula rationality tosome extent.2. The study declared single drug had more significant influence on the mRNA level ofCYP than Composite Salvla Recipe. Composite Salvla Recipe and its Single drugsmight protect heart, and its effect on CYP2and CYP4families might be bilateral.3. These results indicated that PNS at higher concentrations has potentioalhepatotoxicity. PNS had different effects on the mRNA level of CYP in differentconcentrations. It might protect heart and blood vessel, and its effect on CYP2andCYP4families might be bilateral. Doses of medication should be more cared whenPNS was used in clinic.4. Artemisinin can prevent or reduce colonic inflammation, in this case by inducingcytochrome P450expression by activation of PXR. |
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