2012 1, 19(1): 154 160 Journal of Fishery Sciences of China 研究论文 DOI: 10.3724/SP.J.1118.2012.00154 房文红 1, 周常 1,2, 孙贝贝 1, 李国烈 1, 杨先乐 2, 李新苍 1 1, 胡琳琳 1., 200090; 2., 201306 摘要 : (baicalin, BL) (glycyrrhizin, GZ) (Carassius auratus gibelio), (enrofloxacin, ENR) CYP1A CYP3A 7 d (100 mg/kg) (100 mg/kg), 24 h 10 (10 mg/kg),, HPLC (cipmfloxacin, CIP), ; 6 CYP1A CYP3A : (1) (BL) (GZ) (C max ), (AUC) ; (2) BL GZ, (t 1/2z ) (P<0.05), (CLz/F), ; (3) BL GZ, BL GZ C max-cip /C max-enr 1.48% 2.22%, 0.95%; BL GZ AUC 0-t-CIP /AUC 0-t-ENR 2.16% 1.76%, 1.7% C max-cip /C max-enr AUC 0-t-CIP /AUC 0-t-ENR, N- ; (4), BL GZ 7- -O- (EROD, CYP1A ) -N- (ERND, CYP3A ) (P<0.05), CYP1A CYP3A CIP, CYP1A CYP3A : ; ; ; ; ; CYP450 中图分类号 : S94 文献标志码 : A 文章编号 : 1005 8737 (2012)01 0154 07,, [1], (drugdrug interactions, DDIs) DDIs,, DDIs 40% [2], DDIs P450 (CYP450),, CYP450 [3], CYP1A2 [4] ; CYP3A4, [5] (baicalin, BL) (glycyrrhizin, GZ), [6] [7],,, [8] ;, C max (AUC), [9], 收稿日期 : 2011 05 11; 修订日期 : 2011 06 25. 基金项目 : (201203085); ( )(2007M06). 作者简介 :,,.Tel:021-65699301 E-mail: whfang06@yahoo.com.cn
1 : 155,,, [10], P450 (Carassius auratus gibelio),,, 1 1.1 ( 98%) ( ) ; (Enrofloxacin, ENR) (Cipmfloxacin, CIP) 99.5%, Sigma ; ( 98.0%) 7- Sigma, Ⅱ (NADPH) (HPLC ), ( ) 1.2, Waters 2695, Zorbax SB-C 18 (5 µm, 150 4.6 mm, I.D.), Waters 2475 (Beckman Optima L-100XP), (, 970CRT), (, UV-2802S), ( 80, Thermo), (, Millipore Advantage A10), (, Mettler Toledoab 204, 0.000 1 g), (, HITACHI CF16RXⅡ), 0.22 μm 1.3, (200±13) g, 5 35 cm 80 cm 45 cm,, 26~27,, 4 100% 1.4, 0.1 mol/l, 10 mg/ml 3, 20, (BL ) (GZ ), 100 mg/(kg d), 7 d, 1, 5 min 24 h, 6 ;, BL GZ 10, 10 mg/kg, 20 cm 40 cm 30 cm, 5 min 30 min 1 h 2 h 4 h 8 h 12 h 1 d 2 d 3 d 4 d 0.3 ml, 1.5mL, 8 000 r/min 10 min, 80 1.5,, 200 μl 1.5 ml,, 2 min, 10 000 r/min 5 min, 0.22 μm, HPLC 1.6 : 0.01 mol/l ( ph=2.8)=5:95(v/v), 10 min; 40 ; 1.2 ml/min; 10 µl; E x =280 nm, E m =450 nm 1.7 Novi [11] ; 7- -O- (EROD) Lange [12] ; -N- (ERND) Tu [13]
156 19 1.8, DAS 2.1.1 2 2.1 ENR 1 BL ENR, (T max ) (C max ) 0.5 h (7.685±2.392) mg/l; GZ ENR 1, T max =0.083 h, ENR (C max ) (5.769±1.164) mg/l (10.470± 2.088) mg/l BL GZ ENR C max 76% 55% 1 - Fig.1 Mean plasma ENR concentration vs. time curves of crucian carp after coadministration of BL and GZ 2.2 ENR, 1 BL GZ (AUC 0-t ) 132.301 mg h/l 136.358 mg h/l, 46.40% 47.81% BL, (t 1/2z ) 25.90 h, GZ, 30.21 h, (37.38 h); BL GZ (CL z/f ), 2 2.3 CIP 2 3 CIP,, GZ T max, 8 h, C max, (0.099± 0.029) mg/l (0.128±0.024) mg/l; BL, T max =12 h, C max (0.114±0.028) mg/l BL GZ C max-cip /C max-enr 1.48% 2.22%, 0.95% 2.4 CIP, ENR CIP, 2 BL GZ (MRT) (V z/f ) t 1/2z, (CL z/f ) BL GZ CIP (AUC), BL GZ AUC 0-t-CIP /AUC 0-t-ENR 2.16% 1.76%, 1.7% 2.5 P450 BL GZ, CYP1A CYP3A 3 BL EROD ERND (P<0.05), GZ EROD ERND (P<0.01),, 2 1 3 P450, P450,, 7 d CYP1A [14], 14 d CYP3A [15], LS174T CYP3A4 [16] [17] (Paralichthys olivaceus) CYP1A, CYP1A,, EROD ERND CYP1A CYP3A,
1 : 157 Tab.1 表 1 腹腔注射给药后恩诺沙星在异育银鲫血浆中药动学参数 ( 统计矩原理 ) Pharmacokinetic parameters of enrofloxacin in plasma from crucian carp after intraperitoneal injection based on statistical moment theory n=10 pharmacokinetic parameter control baicalin glycyrrhizin C max /(mg L 1 ) 10.470±2.088 7.685±2.392 5.769±1.164 T max /h 0.083 0.5 0.083 AUC 0-t /(mg h L 1 ) 285.142 132.301 136.358 AUC 0- /(mg h L 1 ) 339.674 143.585 152.555 MRT 0-t /h 30.867 26.305 30.500 MRT 0- /h 49.984 34.720 42.084 V z/f /(L kg 1 ) 1.588 2.603 2.858 t 1/2z /h 37.384 25.904 30.217 CL z/f /(L h 1 kg 1 ) 0.029 0.070 0.066 : C max ; T max ; AUC 0-t AUC 0-0-t 0- ; MRT 0-t MRT 0-0-t 0- ; V z/f ; t 1/2z ; CL z/f. Note: C max -peak OTC concentration; T max -time to reach peak concentration; AUC 0-t -area under the concentration-time curve from zero to time t; AUC 0- -area under the concentration-time curve from zero to infinity; MRT 0-t area residence time from zero to time t; MRT 0- area residence time from zero to infinity;v z/f -the apparent volume of distribution; t 1/2z -elimination half-life; CL z/f -total body clearance. 2 - Fig.2 Mean plasma CIP concentration vs. time curves of crucian carp after coadministration of BL and GZ 7 d,, P450 [18],,, [19], C max, BL GZ C max-cip /C max-enr 1.48% 2.22%, 0.95%; BL GZ AUC 0-t-CIP / AUC 0-t-ENR 2.16% 1.76%, 1.7% C max C max-cip /C max-enr AUC 0-t-CIP /AUC 0-t-ENR, N- Vaccaro [19] (Dicentrarchus labrax) P450, NADPH, P450, N-, CYP1A CYP3A, X (PXR), CYP3A, CYP3A N- [15], CYP1A P450 N-,, CYP [20],, CYP1A CYP3A N-,
158 19 Tab.2 表 2 腹腔注射给药后代谢产物环丙沙星在异育银鲫血浆中药动学参数 ( 统计矩原理 ) Pharmacokinetic parameters of ciprofloxacin in plasma from crucian carp after intraperitoneal injection based on statistical moment theory n=10 pharmacokinetic parameter control baicalin glycyrrhizin C max /(mg L 1 ) 0.099±0.029 0.114±0.028 0.128±0.024 T max /h 8 12 8 AUC 0-t /(mg h L 1 ) 5.022 2.859 2.400 AUC 0- /(mg h L 1 ) 7.567 2.906 2.422 MRT 0-t /h 38.108 21.564 15.192 MRT 0- /h 86.388 23.151 16.160 V z/f /(L kg 1 ) 113.179 83.359 93.906 t 1/2z /h 59.350 16.786 15.764 CL z/f /(L h 1 kg 1 ) 1.322 3.441 4.128 : C max ; T max ; AUC 0-t AUC 0-0-t 0- ; MRT 0-t MRT 0-0-t 0- ; V z/f ; t 1/2z ; CL z/f. Note: C max peak OTC concentration; T max time to reach peak concentration; AUC 0-t area under the concentration-time curve from zero to time t; AUC 0- area under the concentration-time curve from zero to infinity; MRT 0-t area residence time from zero to time t; MRT 0- area residence time from zero to infinity; V z/f apparent volume of distribution; t 1/2z elimination half-life; CL z/f total body clearance. Tab. 3 表 3 BL 和 GZ 对异育银鲫肝 CYP1A 和 CYP3A 活性的影响 Effects of BL and GZ on CYP1A and CYP3A activities of liver from crucian carp 1 n=6; x ±SD; pmol min 1 mg dose/ EROD ERND group (mg kg 1 d 1 ) EROD activity ERND activity baicalin 100 52.65±2.64 * 2185.72±207.11 ** glycyrrhizin 100 51.43±3.99 * 2108.50±98.96 ** control 0 35.4±3.6 1166.47±125.71 : * (P<0.05), ** (P<0.01). Note: *donates singnificant difference with control (P<0.05). **donates extremely singnificant difference with control (P<0.01). C max AUC 0-t (t 1/2z ) (CL z/f ),,,, CYP1A2, [4] ;, [8] ; A t 1/2,, AUC, [21] ;, A AUC C max A t 1/2, [22], P450 ( CYP3A), P- P-, ATP (ABC),, [23] Bauer [24] (Hypericum perforatum) P- A [25] [26] P-, [25 26] P-,,
1 : 159 (pefloxacin) [27], (danofloxacin) [28] ; Seelig [29] P-,, P- P-, P-, ENR 73, 27, [30],,,, 参考文献 : [1]. [J]., 2003,1(4): 351 352. [2],,. - [J]. :, 2009, 28(2): 215 224. [3],,. P450 [J]., 2011, 38(1): 52 57. [4] Kubota Y, Kobayashi K, Tanaka N, et al. Pretretment with Ginkgo biloba extract weakens the hypnosis action of Phenobarbital and its plasma concentration in rats [J]. Pharmacology, 2004, 56(3): 401 405. [5] Harkey M R, Henderson G L, Gershwin M E, et al. Variability in commercial ginseng products: An analysis of 25 preparations [J]. Am J Clin Nutr, 2001, 73: 1101 1106. [6] Isbrucker R A, Burdock G A. Risk and safety assessment on the consumption of Licorice root (Glycyrrhiza sp.), its extract and powder as a food ingredient, with emphasis on the pharmacology and toxicology of glycyrrhizin [J]. Regul- Toxicol Pharmacol, 2006, 46: 167 192. [7] Chang H H, Yi P L, Cheng C H, et al. Biphasic effects of baicalin, an active constituent of Scutellaria baicalensis Georgi, in the spontaneous sleep-wake regulation [J]. J Ethnopharmacol, 2011, 135(2): 359 368. [8] Velpandian T, Jasuja R, Bhardwaj R K, et al. Piperine in food: interference in the pharmacokinetics of phenytoin [J]. Eur J Drug M etab Pharmacokinet, 2001, 26(4): 241 247 [9],,,. LC-MS/MS [J]., 2010, 30(9): 1664 1671. [10]. [M]., 2008: 53,173. [11] Novi S, Pretti C, Cognetti A M, et al. Biotransformation enzymes and their induction by β-naphtoflavone in adult sea bass (Dicentrarchus labrax) [J]. Aquat Toxicol, 1998, 41: 63 81. [12] Lange U, Goksøyr A, Siebers D, et al. Cytochrome P450 1A-dependent enzyme activities in the liver of dab (Limanda limanda): kinetics, seasonal changes and detection limits [J]. Comp Biochem Physiol, 1999, 123B: 361 371. [13] Tu Y Y, Yang C S. High-affinity nitrosamine dealkylase system in rat liver microsomes and its induction by fasting [J]. Cancer Res, 1983, 43: 623 629. [14],,,. P4501A2 [J]., 2007, 7(4): 278 280. [15]. CYP450 [D]. :, 2010. [16] Li Y, Wang Q, Yao X M, et al. Induction of CYP3A4 and MDR1 gene expression by baicalin, baicalein, chlorogenic acid, and ginsenoside Rf through constitutive androstane receptor-and pregnane X receptor-mediated pathways [J]. Eur J Pharmacol, 2010, 640: 46 54. [17],,,. CYP1A [J]., 2010, 17(5): 1121 1127. [18],,,. P450 [J]., 2011, 27(4): 519 523. [19] Vaccaro E, Giorgi M, Longo V, et al. Inhibition of cytochrome P450 enzymes by enrofloxacin in the sea bass (Dicentrarchus labrax) [J]. Aquat Toxicol, 2003, 62:27 33. [20] Yoo H H, Kim N S, Lee J, et al. Characterization of human cytochrome P450 enzymes involved in the biotransformation of eperisone [J]. Xenobiotica, 2009, 39: 1 10. [21],,,. [J]., 2002, 16(1): 12 15. [22],,,. A [J]., 2006, 41( 9): 882 887. [23] Schinkel A H, Jonker J W. Mammalian drug efflux transporters of the ATP binding cassette (ABC) family: an overview [J]. Adv Drug Deliv Rev, 2003,55(1): 3 29.
160 19 [24] Bauer S, Stormer E, Johne A, et al. Alterations in cyclosporin A pharmacokinetics and metabolism during treatment with St John s wort in renal transplant patients[j]. Br J Clin Pharmacol, 2003, 55:203 211. [25]. CYP450 [D]. :, 2008. [26]. 18 P- [D].. :, 2010. [27] Seelig A, Landwojtowicz E. Structure activity relationship of P-glycoprotein substrates and modifiers [J]. Eur J Pharmacol Sci, 2000, 12:31 40. [28] Schrickx J A, Gremmels J F. Danofloxacin-mesylate is a substrate for ATP-dependent efflux transporters [J]. Br J Pharmacol, 2007, 150(4):463 469. [29] Seelig A, Landwojtowicz E. Structure activity relationship of P-glycoprotein substrates and modifiers [J]. Eur J Pharmaceutical Sci, 2000, 12:31 40. [30]. [M]. :, 2008. Effect of baicalin and glycyrrhizin on enrofloxacin in vivo in crucian carp (Carassius auratus gibelio) FANG Wenhong 1, ZHOU Chang 1.2, SUN Beibei 1, LI Guolie 1, YANG Xianle 2, LI Xincang 1, HU Linlin 1 1. East China Sea Fisheries Research Institute, Chinese Academy of Fishery sciences; Key Laboratory of Marine and Estuarine Fisheries Resources and Ecology, Ministry of Agriculture, Shanghai 200090 China; 2. National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai 201306, China Abstract: Cytochrome P450s (CYPs) widely distribute in the liver of aquatic organisms and play a key role in drug metabolism. CYPs catalyze the N-deethylation of Enrofloxacin (ENR) to Ciprofloxacin (CIP). However, there is limited information regarding the action of CYPs in fish. We evaluated the metabolism of ENR following oral administrated of baicalin (100 mg/kg) and glycyrrhizin (100 mg/kg) in crucian carp, Carassium auratus gibelio. We measured the enzymatic activity of cytochrome P450 1A (CYP1A) and 3A (CYP3A) in liver microsomes. In addition, we documented the pharmacokinetics of ENR by continuous blood sampling. The absorption of ENR decreased significantly over time and both C max and AUC also deceased. AUC and t 1/2z decreased signficantly whereas CL inceased after oral administrated of baicalin and glycyrrhizin, suggesting that ENR elimination was accelerated. The C max CIP /C max ENR was 1.48% and 2.22% for the BL and GZ groups, respectively (control: 0.95%). The AUC 0-t-CIP /AUC 0-t-ENR was 2.16% and 1.76% for the BL and GZ groups, respectively (control: 1.7%). Thus, BL and GZ contributed to induce N-deethylation of ENR. Pretreatment with BL and GZ was associated with a significant (P<0.05) incease in ethoxyresorufin-o-deethylation (EROD) and erythromycin-n-demethylation (ERND) activity, the specific probes for CYP1A and CYP3A, respectively. In summary, BL and GZ accelerated the elimination of ENR. Furthermore, production of the ENR metabolite was related to the induction of CYP1A and CYP3A enzymatic activity. Key words: Carassium auratus gibelio; enrofloxacin; pharmacokinetics; baicalin; glycyrrhizin; CYP450 Corresponding author: FANG Wenhong. Tel:+86-21-65699301; E-mail: whfang06@yahoo.com.cn