三七总皂苷对细胞色素P450酶CYP3A4和CYP2C9的影响
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摘要
目的探讨三七总皂苷对Chang Liver细胞P450酶(CYP450) CYP3A4和CYP2C9表达的影响。方法采用CCK-8法考察三七总皂苷对Chang Liver细胞活力的影响,确定给药剂量范围;采用定量RT-PCR法考察三七总皂苷在给药后4,8,24,48 h对Chang Liver细胞中CYP3A4和CYP2C9 mRNA表达的影响;通过蛋白质印迹(Western Blot)法考察三七总皂苷在给药后4,8,24,48 h对Chang Liver细胞中CYP3A4和CYP2C9蛋白表达的影响。结果 100μg/m L三七总皂苷干预Chang Liver细胞4,8,24,48 h后,各干预组CYP2C9蛋白表达量明显高于空白对照组(P <0. 05),各组诱导水平随着作用时间的延长相应增加。8 h组、24 h组、48 h组CYP3A4蛋白表达量明显高于空白对照组(P <0. 05),各组的相对蛋白表达量呈时间依赖性,8 h组、24 h组、48 h组CYP2C9和CYP3A4的mRNA表达量均明显高于空白对照组(P <0. 05),且各组的诱导水平随着作用时间的延长相应增加。结论三七总皂苷可诱导CYP3A4和CYP2C9的mRNA及蛋白的表达,临床在使用三七总皂苷时,需要关注其与CYP3A4和CYP2C9底物药物之间可能的相互作用。
Objective To investigate the effect of panax notoginseng saponins(PNS) on the expression of cytochrome P450 enzymes(CYP 450)CYP3A4 and CYP2C9 in Chang Liver cells. Methods The effect of PNS on the viability of Chang Liver cells were investigated by CCK-8 assay,and the dosage range was determined. The effect of PNS on the expression of CYP3A4 and CYP2C9 mRNA in Chang Liver cells were investigated by quantitative RT-PCR method at 4, 8, 24 and 48 h after administration. The effect of PNS on the expression of CYP3A4 and CYP2C9 protien in Chang Liver cells were investigated by Western Blot method at 4,8,24 and 48 h after administration. Results Compared with the blank control group,the expression of CYP2C9 protein in Chang Liver cells was significantly higher after 4,8,24 and 48 h of treatment with 100 μg/mL PNS( P < 0. 05), and the induction level of each group increased with the prolongation of the action time. The expression of CYP3A4 protein in 8 h group,24 h group and 48 h group was significantly higher than that in the blank control group( P < 0. 05),and the relative expression of CYP3A4 protein in each group was time-dependent.The expression of CYP2C9 and CYP3A4 mRNA in 8 h group, 24 h group and 48 h group were significantly higher than that in the blank control group( P < 0. 05), and the induction level of each group increased with the prolongation of action time. Conclusion PNS can induce the expression of CYP3A4 and CYP2C9 mRNA and protien. It is necessary to pay attention to the possible drug-drug interaction between PNS and CYP3A4 and CYP2C9 substrates in clinic.
Objective To investigate the effect of panax notoginseng saponins(PNS) on the expression of cytochrome P450 enzymes(CYP 450)CYP3A4 and CYP2C9 in Chang Liver cells. Methods The effect of PNS on the viability of Chang Liver cells were investigated by CCK-8 assay,and the dosage range was determined. The effect of PNS on the expression of CYP3A4 and CYP2C9 mRNA in Chang Liver cells were investigated by quantitative RT-PCR method at 4, 8, 24 and 48 h after administration. The effect of PNS on the expression of CYP3A4 and CYP2C9 protien in Chang Liver cells were investigated by Western Blot method at 4,8,24 and 48 h after administration. Results Compared with the blank control group,the expression of CYP2C9 protein in Chang Liver cells was significantly higher after 4,8,24 and 48 h of treatment with 100 μg/mL PNS( P < 0. 05), and the induction level of each group increased with the prolongation of the action time. The expression of CYP3A4 protein in 8 h group,24 h group and 48 h group was significantly higher than that in the blank control group( P < 0. 05),and the relative expression of CYP3A4 protein in each group was time-dependent.The expression of CYP2C9 and CYP3A4 mRNA in 8 h group, 24 h group and 48 h group were significantly higher than that in the blank control group( P < 0. 05), and the induction level of each group increased with the prolongation of action time. Conclusion PNS can induce the expression of CYP3A4 and CYP2C9 mRNA and protien. It is necessary to pay attention to the possible drug-drug interaction between PNS and CYP3A4 and CYP2C9 substrates in clinic.
引文
[1]方鹏飞,常丽霞,宋渊.三七总皂苷临床应用研究进展[J].中医药学报,2016,44(3):120-123.
[2] LIAO J,WEI B,CHEN H,et al. Bioinformatics investigation of therapeutic mechanisms of Xuesaitong capsule treating ischemic cerebrovascular rat model with comparative transcriptomeanalysis[J].Am J Transl Res,2016(8):2438-2449.
[3]何三民,余志怡,戴德雄.血塞通泡腾片中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量测定[J].中国药业,2011,20(11):30-31.
[4] TRACY TS,CHAUDHRY AS,PRASAD B,et al. Interindividual Variability in Cytochrome P450-Mediated Drug Metabolism[J].Drug Metab Dispos,2016,44(3):343-351.
[5] WU JJ,AI CZ,LIU Y,et al. Interactions between phytochemicals from traditional Chinese medicines and human cytochrome P450enzymes[J]. Curr Drug Metab,2012,13(5):599-614.
[6]魏春燕,吴逢波,徐. CYP450与药物相互作用[J].中国药业,2014,23(6):17-20.
[7]王常顺,刘永利,王晓蕾,等. UPLC、HPLC测定血栓通注射液中5个皂苷类成分含量的比较研究[J].药物分析杂志,2013,33(9):1617-1620.
[8]王莹,褚扬,李伟,等.三七中皂苷成分及其药理作用的研究进展[J].中草药,2015,46(9):1381-1392.
[9]杨琳,林万程,施家乐.三七总皂苷药理作用的研究进展[J].组别安徽医药,2014(5):963-965.
[10] GERTZ M,CARTWRIGHT CM,HOBBS MJ,et al. Cyclosporine inhibition of hepatic and intestinal CYP3A4,uptake and efflux transporters:application of PBPK modeling in the assessment of drug-drug interaction potential[J]. Pharm Res,2013,30(3):761-780.
[11] KOMURA H,IWAKI M. In vitro and in vivo small intestinal metabolism of CYP3A and UGT s ubstrates in preclinical animals species and humans:species differences[J]. Drug Metab Rev,2011,43(4):476-498.
[12]陈艳进,王宇光,马增春,等.三七总皂苷对大鼠肝脏药物代谢酶酶活性、mRNA及蛋白表达的影响[J].中国中药杂志,2014,39(19):3824-3828.
[13] HAN M,FU S,FANG XL,et al. Comparison between the charateristics of absorption and pharmacokinetic behavior of ginsenoside Rg1 and ginsenoside Rb,of Panax notoginseng saponins[J].Yao Xue Xue Bao,2007,42(8):849-853.
[14] LIU R,QIN MN,HANG PZ,et al. Effects of Panax notoginseng Saponins on the Activities of CYP1A2,CYP2C9,CYP2D6and CYP3A4 in Rats In Vivo[J]. Phytotherapy Research,2012,26(8):1113-1118.
[15]石杰,陈安进.血塞通及川芎嗪对细胞色素P450不同亚型代谢酶影响的研究[J].中国中西医结合急救杂志,2008,15(6):342-345.
[16]马维娜,吕磊,孟拥军. HPLC-MS法测定大鼠体内血塞通对氯沙坦钾的药动学影响[J].中国药师,2017,20(11):1937-1941.
[17] WANG R,ZHANG H,WANG YJ,et al. Effects of salvianolic acid B and tanshinoneⅡA on the pharmacokinetics of Losartan in rats by regulating the activities and expression of CYP3A4 and CYP2C9[J]. J Ethnopharmacol,2016,180:87-96.
[2] LIAO J,WEI B,CHEN H,et al. Bioinformatics investigation of therapeutic mechanisms of Xuesaitong capsule treating ischemic cerebrovascular rat model with comparative transcriptomeanalysis[J].Am J Transl Res,2016(8):2438-2449.
[3]何三民,余志怡,戴德雄.血塞通泡腾片中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量测定[J].中国药业,2011,20(11):30-31.
[4] TRACY TS,CHAUDHRY AS,PRASAD B,et al. Interindividual Variability in Cytochrome P450-Mediated Drug Metabolism[J].Drug Metab Dispos,2016,44(3):343-351.
[5] WU JJ,AI CZ,LIU Y,et al. Interactions between phytochemicals from traditional Chinese medicines and human cytochrome P450enzymes[J]. Curr Drug Metab,2012,13(5):599-614.
[6]魏春燕,吴逢波,徐. CYP450与药物相互作用[J].中国药业,2014,23(6):17-20.
[7]王常顺,刘永利,王晓蕾,等. UPLC、HPLC测定血栓通注射液中5个皂苷类成分含量的比较研究[J].药物分析杂志,2013,33(9):1617-1620.
[8]王莹,褚扬,李伟,等.三七中皂苷成分及其药理作用的研究进展[J].中草药,2015,46(9):1381-1392.
[9]杨琳,林万程,施家乐.三七总皂苷药理作用的研究进展[J].组别安徽医药,2014(5):963-965.
[10] GERTZ M,CARTWRIGHT CM,HOBBS MJ,et al. Cyclosporine inhibition of hepatic and intestinal CYP3A4,uptake and efflux transporters:application of PBPK modeling in the assessment of drug-drug interaction potential[J]. Pharm Res,2013,30(3):761-780.
[11] KOMURA H,IWAKI M. In vitro and in vivo small intestinal metabolism of CYP3A and UGT s ubstrates in preclinical animals species and humans:species differences[J]. Drug Metab Rev,2011,43(4):476-498.
[12]陈艳进,王宇光,马增春,等.三七总皂苷对大鼠肝脏药物代谢酶酶活性、mRNA及蛋白表达的影响[J].中国中药杂志,2014,39(19):3824-3828.
[13] HAN M,FU S,FANG XL,et al. Comparison between the charateristics of absorption and pharmacokinetic behavior of ginsenoside Rg1 and ginsenoside Rb,of Panax notoginseng saponins[J].Yao Xue Xue Bao,2007,42(8):849-853.
[14] LIU R,QIN MN,HANG PZ,et al. Effects of Panax notoginseng Saponins on the Activities of CYP1A2,CYP2C9,CYP2D6and CYP3A4 in Rats In Vivo[J]. Phytotherapy Research,2012,26(8):1113-1118.
[15]石杰,陈安进.血塞通及川芎嗪对细胞色素P450不同亚型代谢酶影响的研究[J].中国中西医结合急救杂志,2008,15(6):342-345.
[16]马维娜,吕磊,孟拥军. HPLC-MS法测定大鼠体内血塞通对氯沙坦钾的药动学影响[J].中国药师,2017,20(11):1937-1941.
[17] WANG R,ZHANG H,WANG YJ,et al. Effects of salvianolic acid B and tanshinoneⅡA on the pharmacokinetics of Losartan in rats by regulating the activities and expression of CYP3A4 and CYP2C9[J]. J Ethnopharmacol,2016,180:87-96.