肾康注射液对大鼠药代动力学及细胞色素2C9酶活性的影响
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摘要
目的以双氯芬酸为探针底物探讨肾康注射液对大鼠体内细胞色素2C9(CYP2C9)酶活性的影响。方法将24只SD雄性大鼠随机分为对照组,低、中、高剂量肾康注射液组,每组6只,分别腹腔注射0.9%NaCl和6.7,13.3,26.6m L·kg~(-1)肾康注射液,每日2次,均连续给药7 d。第8天收集血浆样品前,在给药1次、给药1 h后,各组股静脉注射双氯芬酸注射液15 mg·kg~(-1)。分别于双氯芬酸给药前及给药后5,15,30,45 min和1,1.5,2,3,4,6,8,10,12,24 h眼底静脉丛采集血浆样品。用高效液相色谱法测定血浆中双氯芬酸及其代谢物4'-羟基双氯芬酸的血药浓度,并用Winnolin 6.1药代动力学软件中非房室模型拟合并计算药代动力学参数。结果对照组与低、中、高3个剂量肾康注射液组的双氯芬酸的AUC_(0→24 h)分别为(21.12±2.88),(25.13±8.13),(31.11±11.19),(32.32±6.18)mg·h·L~(-1),与对照组比较,差异无统计学意义(P>0.05);对照组与低、中、高3个剂量肾康注射液组的4'-羟基双氯芬酸的AUC_(0→24 h)分别为(9.79±1.12),(7.84±2.67),(9.97±3.85),(10.39±3.38)mg·h·L~(-1)(P>0.05);对照组与低、中、高3个剂量肾康注射液组的双氯芬酸的t1/2分别为(6.21±1.78);(7.46±2.20);(6.40±1.50),(6.13±2.19)h(P>0.05);4'-羟基双氯芬酸的t1/2分别为(11.53±4.31);(30.85±10.52),(23.29±13.09),(22.53±10.95)h(P>0.05);其他药代动力学参数差异均无统计学意义(P>0.05)。结论连续给药肾康注射液很可能不影响CYP2C9典型探针底物双氯酚酸的代谢,因此肾康注射液很可能不影响大鼠CYP2C9酶活性。
Objective Diclofeanc as probe drug to investigate the influence of Shenkang injection on the activity of CYP2C9 enzyme in rats.Methods A total of 24 SD male rats were randomly divided into control group( 0. 9% NaCl),low dose( 6. 7 mL · kg~(-1)),middle dose( 13. 3m L·kg~(-1)),high dose( 26. 6 mL · kg~(-1)) Shenkang injection group,each group 6 rats. The relevant injection was administered by intraperitoneal injection,twice a day for 7 d. On the eighth day before collected the blood plasma samples each group were given relevant injection and all rats were injected with diclofenac( 15 mg·kg~(-1),iv) 1 h after last administration. The orbital blood samples were collected before and 5,15,30,45 min and 1,1. 5,2,3,4,6,8,10,12,24 h after diclofenac administration through femoral venous. A sensitive high-performance liquid chromatography method was established to characterize andvalidate for the determination of plasma concentration of diclofenac and 4'-hydroxy-diclofenac in rats. Pharmacokinetic parameters were calculated with Win Nolin 6. 1 pharmacokinetic software. Results The area under the concentration-time curve( AUC_(0→24 h)) of diclofenac in control group,low,middle and high-dose Shenkang injection group were( 21. 12 ± 2. 88),( 25. 13 ± 8. 13),( 31. 11 ± 11. 19),( 32. 32 ± 6. 18) mg · h · L~(-1)( P > 0. 05). The AUC_(0→24 h)of 4'-hydroxy-diclofenac were( 9. 79 ± 1. 12),( 7. 84 ± 2. 67),( 9. 97 ± 3. 85),( 10. 39 ± 3. 38) mg·h·L~(-1)( P >0. 05). The t1/2 of diclofenac in control group,low,middle and high-dose Shenkang injection group were( 6. 21 ± 1. 78),( 7. 46 ± 2. 20),( 6. 40 ± 1. 50),( 6. 13 ± 2. 19) h( P > 0. 05). The t1/2 of 4'-hydroxy-diclofenac in control group,low,middle and high-dose Shenkang injection group were( 11. 53 ± 4. 31),( 30. 85 ± 10. 52),( 23. 29 ± 13. 09),( 22. 53 ± 10. 95) h( P > 0. 05). There was no statistical significant difference in other pharmacokinetic parameters( P > 0. 05). Conclusion Consecutive administration of Shenkang injection may not have an effect on the pharmacokinetic of diclofenac and 4'-hydroxy-diclofenac of typical probe substrates of CYP2C9 in rats. The present study would provide some reference on Shenkang injection have no influence on CYP2C9.
Objective Diclofeanc as probe drug to investigate the influence of Shenkang injection on the activity of CYP2C9 enzyme in rats.Methods A total of 24 SD male rats were randomly divided into control group( 0. 9% NaCl),low dose( 6. 7 mL · kg~(-1)),middle dose( 13. 3m L·kg~(-1)),high dose( 26. 6 mL · kg~(-1)) Shenkang injection group,each group 6 rats. The relevant injection was administered by intraperitoneal injection,twice a day for 7 d. On the eighth day before collected the blood plasma samples each group were given relevant injection and all rats were injected with diclofenac( 15 mg·kg~(-1),iv) 1 h after last administration. The orbital blood samples were collected before and 5,15,30,45 min and 1,1. 5,2,3,4,6,8,10,12,24 h after diclofenac administration through femoral venous. A sensitive high-performance liquid chromatography method was established to characterize andvalidate for the determination of plasma concentration of diclofenac and 4'-hydroxy-diclofenac in rats. Pharmacokinetic parameters were calculated with Win Nolin 6. 1 pharmacokinetic software. Results The area under the concentration-time curve( AUC_(0→24 h)) of diclofenac in control group,low,middle and high-dose Shenkang injection group were( 21. 12 ± 2. 88),( 25. 13 ± 8. 13),( 31. 11 ± 11. 19),( 32. 32 ± 6. 18) mg · h · L~(-1)( P > 0. 05). The AUC_(0→24 h)of 4'-hydroxy-diclofenac were( 9. 79 ± 1. 12),( 7. 84 ± 2. 67),( 9. 97 ± 3. 85),( 10. 39 ± 3. 38) mg·h·L~(-1)( P >0. 05). The t1/2 of diclofenac in control group,low,middle and high-dose Shenkang injection group were( 6. 21 ± 1. 78),( 7. 46 ± 2. 20),( 6. 40 ± 1. 50),( 6. 13 ± 2. 19) h( P > 0. 05). The t1/2 of 4'-hydroxy-diclofenac in control group,low,middle and high-dose Shenkang injection group were( 11. 53 ± 4. 31),( 30. 85 ± 10. 52),( 23. 29 ± 13. 09),( 22. 53 ± 10. 95) h( P > 0. 05). There was no statistical significant difference in other pharmacokinetic parameters( P > 0. 05). Conclusion Consecutive administration of Shenkang injection may not have an effect on the pharmacokinetic of diclofenac and 4'-hydroxy-diclofenac of typical probe substrates of CYP2C9 in rats. The present study would provide some reference on Shenkang injection have no influence on CYP2C9.
引文
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[7]DIEHL K H,HULL R,MORTON D,et al.A good practice guide to the administration of substances and removal of blood,including routes and volumes[J].J Appl Toxicol,2001,21(1):15-23.
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[2]QIU F,ZHANG R,SUN J,et al.Inhibitory effects of seven components of danshen extract on catalytic activity of cytochrome P450 enzyme in human liver microsomes[J].Drug Metab Dispos,2008,36(7):1308-1314.
[3]TANG J C,YANG H,SONG X Y,et al.Inhibition of cytochrome P450 enzymes by rhein in rat liver microsomes[J].Phytother Res,2009,23(2):159-164.
[4]LEE B,JI H K,LEE T,et al.Simultaneous screening of activities of five cytochrome P450 and four uridine 5'-diphospho-glucuronosyltransferase enzymes in human liver microsomes using cocktail incubation and liquid chromatography-tandem mass spectrometry[J].Drug Metab Dispos,2015,43(7):1137-1146.
[5]GERIN B,DELL'AIERA S,RICHERT L,et al.Assessment of cytochrome P450(1A2,2B6,2C9 and 3A4)induction in cryopreserved human hepatocytes cultured in 48-well plates using the cocktail strategy[J].Xenobiotica,2013,43(4):320-335.
[6]汪电雷,刘李,邓远雄,等.银杏内酯B对CYP2C9底物双氯芬酸在大鼠体内药代动力学和药效学的影响[J].中国新药杂志,2008,17(11):919-922.
[7]DIEHL K H,HULL R,MORTON D,et al.A good practice guide to the administration of substances and removal of blood,including routes and volumes[J].J Appl Toxicol,2001,21(1):15-23.
[8]KASPEREK R,ZIMMER L,JAWIEN W,et al.Pharmacokinetics of diclofenac sodium and papaverine hydrochloride after oral administration of tablets to rabbits[J].Acta Pol Pharm,2015,72(3):527-538.