氟喹诺酮类药物对肝外药物代谢酶及GR、GSH-Px活性的影响
|
摘要
目的：探讨氟喹诺酮类药物（fluoroquinolones，FQs）对大鼠肺、脑、肾、小肠中细胞色素P-450（cytochrome P-450，P-450）含量、氨基比林N-脱甲基酶（aminopyrine N-demethylase，AMND）、红霉素N-脱甲基酶（erythromycin N-demethylase，ERND）、谷胱甘肽S-转移酶（glutathione S-transferase，GST）、谷胱甘肽还原酶（glutathione reductase，GR）、谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）活性的影响。方法：实验分为体外实验和体内实验。体外实验中，用8只大鼠的肺、脑、肾、小肠制备S9（postmitochondria supernatant），分别测定反应体系中加入环丙沙星（ciprofloxacin，CPFX）、妥苏沙星（tosufloxacin，TFLX）、司帕沙星（sparfloxacin，SPFX）前后的酶活性和P-450含量，药物组反应体系中各药物的终浓度为1mmol/L。体内实验中，将32只大鼠分为对照组和各药物组，分别以0.25%羧甲基纤维素4ml/kg和各药物1mmol/kg灌胃，qd×7d, 末次给药后6小时处死，取肺、脑、肾、小肠制备S9测定酶活性和P-450含量。结果：体内实验和体外实验均能在大鼠的肺、肾组织中测到较低水平的P-450含量, 但各药物组P-450含量与对照组无显著性差异；三种药物对各组织的AMND和GST活性较对照组均有不同程度的抑制（P<0.05），但体外实验与体内实验中各药物组与对照组的差异有所不同，且不同组织中GST活性降低的程度也不同；ERND的活性在肾和小肠组织可测到，在肺、脑中未测到；药物组各组织的GR、GSH-PX和 肾、小肠的ERND活性较对照组有降低，但大多无显著性差异。结论：FQs对肝外组织的AMND、GST活性有一定的抑制作用，且对不同组织的GST活性的抑制作用存在差别，对ERND、GR、GSH-PX有抑制的趋势。
Objective: To explore the effects of fluoroquinolones（FQs） on the content of cytochrome and the activities of aminopyrine N-demethylase（AMND）、erythromycin N-demethylase（ERND）、glutathione S-transferase（GST）, gluathione reductase （GR）and gluathione peroxidase（GSH-PX） in the lung, brain, kidney, and small intestine of rats. Methods: ciprofloxacin, tosufloxacin and sparfloxacin were selected to represent FQs, while carboxymethylcellulose was used as the control. The experiment was divided into two parts, in vitro and in vivo, 8 and 32 wistar rats were used in the research, respectively; lung, brain, kidney and small intestine of rats were took out and made to S9 mix. In vitro test, the rats was killed directly without drug intervention, CPFX、TFLX、SPFX or carboxymethylcellulose were added in the reaction architecture of S9 mix, the concentration of drug in reaction architecture was 1mmol/L; in vivo test, 32 rats were divided into 4 groups, before killing, the rats of 4 groups were continuously perfused with 0.25% carboxy- methylcellulose and CPFX、TFLX 、SPFX （1mmol/kg.weight）respectively for 7 days and one time per day. In the end, the content of P-450 and the activity of above-mentioned enzyme in the S9 mix was assayed with spectrophotometer. Results: In vitro and in vivo, low level of P-450 was detected in lung and kidney tissue, but there is no significant difference between the drug groups and the control group. Activities of AMND and GST of brain were inhibited significantly by CPFX, TFLX and SPFX with contrast to carboxymethycellulose（P<0.05）, but the difference between the drug groups and the control group in vitro are not consistent with the results of in vivo, and the decline of the activity of GST in 4 tissues were not on the same level. The activity of ERND in kidney and small intestine can be detected but very low, and no activity of ERND was determined in lung and brain; meanwhile, even though the activities of ERND,GR and GSH-PX were inhibited by FQs in all 4 tissues, but it did not have significant difference. Conclusions: FQS can inhibit the activities of AMND and GST to some extent in some extrahepatic tissues, and it`s
inhibition to GST in different tissues is not consistent. Meanwhile , there is an inhibition trend of FQs on ERND, GR and GSH-PX activities.
Objective: To explore the effects of fluoroquinolones（FQs） on the content of cytochrome and the activities of aminopyrine N-demethylase（AMND）、erythromycin N-demethylase（ERND）、glutathione S-transferase（GST）, gluathione reductase （GR）and gluathione peroxidase（GSH-PX） in the lung, brain, kidney, and small intestine of rats. Methods: ciprofloxacin, tosufloxacin and sparfloxacin were selected to represent FQs, while carboxymethylcellulose was used as the control. The experiment was divided into two parts, in vitro and in vivo, 8 and 32 wistar rats were used in the research, respectively; lung, brain, kidney and small intestine of rats were took out and made to S9 mix. In vitro test, the rats was killed directly without drug intervention, CPFX、TFLX、SPFX or carboxymethylcellulose were added in the reaction architecture of S9 mix, the concentration of drug in reaction architecture was 1mmol/L; in vivo test, 32 rats were divided into 4 groups, before killing, the rats of 4 groups were continuously perfused with 0.25% carboxy- methylcellulose and CPFX、TFLX 、SPFX （1mmol/kg.weight）respectively for 7 days and one time per day. In the end, the content of P-450 and the activity of above-mentioned enzyme in the S9 mix was assayed with spectrophotometer. Results: In vitro and in vivo, low level of P-450 was detected in lung and kidney tissue, but there is no significant difference between the drug groups and the control group. Activities of AMND and GST of brain were inhibited significantly by CPFX, TFLX and SPFX with contrast to carboxymethycellulose（P<0.05）, but the difference between the drug groups and the control group in vitro are not consistent with the results of in vivo, and the decline of the activity of GST in 4 tissues were not on the same level. The activity of ERND in kidney and small intestine can be detected but very low, and no activity of ERND was determined in lung and brain; meanwhile, even though the activities of ERND,GR and GSH-PX were inhibited by FQs in all 4 tissues, but it did not have significant difference. Conclusions: FQS can inhibit the activities of AMND and GST to some extent in some extrahepatic tissues, and it`s
inhibition to GST in different tissues is not consistent. Meanwhile , there is an inhibition trend of FQs on ERND, GR and GSH-PX activities.
引文
[1] Vijayalakshmi R, Hindupur K, Amamda TV. High activity of cytochrome P-450-linked aminopyrine N-demethylase in mouse brain microsomes, and associated sex-related difference. Biochem. J., 1989;261(3): 769-773
[2] Ueng YF, Shyu CC, Lin YL, et al. Effects of baicalein and wogonin on drug-metabolizing enzymes in C57BL/6J mice. Life-Sci, 2000; 67(18): 2189-2200
[3] Bohnenstengel F, Friedel G, Ritter CA, et al. Variability of cyclophosphamide uptake into human bronchial carcinoma: consequences for local bioactivation. Cancer Chemother Pharmacol. 2000; 45(1): 63-68
[4] Lieber CS. Alcohol and the liver: metabolism of alcohol and its role in hepatic and extrahepatic diseases. Mt Sinai J Med, 2000; 67(1): 84-94
[5] 周义文, 钱元恕, 盛家琦. 环丙沙星对人肝微粒体药酶活性的影响. 现代医药卫生, 2001;17（9）:668-669
[6] Valera F, De La Torre R, Segura J. Selective in vitro inhibition of hepatic oxidative metabolism by quinolones:7-ethyresorufin and caffeine as model substrates. J pharmacol, 1991;43:17
[7] Davis JD, Houston JB. Metabolism of theophylline and its inhibition by fluoroquinolones in rat hepatic microsomes. Xenobiotica, 1995;25(6):563-573
[8] Mayeaux MH, Winston GW. Amtibiotic effects on cytochromes P-450 content and mixed-function oxygenase. J Vet Pharmacol Ther, 1998;21(4):274-281
[9] 莫岚, 钱元恕, 王其南等. 培氟沙星对大鼠肝药酶的抑制作用. 新药与临床, 1995;14 (1):15-17
[10] 莫岚, 钱元恕, 王其南等. 依诺沙星对大鼠肝药酶的抑制作用. 中国抗生素杂志, 1994;19(6):459-463
[11] 黄仁刚, 钱元恕, 兰雁飞等. 洛美沙星对大鼠肝脏混合功能氧化酶的作用. 中国抗生素杂志, 1999;24(1):53-55
[12] Motomura M, Kataoka K, Takeo G, et al. Hippocampus and fromtal cortex are the potemtial mediatory sites for comvulsions induced by new quinolones and non-steroidal anti-inflammatory drugs. Int J Chin Pharmacol Ther Toxicol,1991'29(6):223-227
[13] Peng RX, Wang H, Wang YS, et al. Glutathione-related enzyme activities in human fatal adrenal, liver, and kidney. Acta Pharmacol Sinica, 1998;19(2):167-171
[14] 徐淑云,卞如濂,陈修. 《药理实验方法学》.第二版，北京:人民卫生出版社,1991;488-95
[15] 方治平,宋晓红,刘小康. 临床分离耐氟喹诺酮类金黄色葡萄球菌的gyrA基因单点突变研究. 华夏医科大学学报，2000;31（1）:24-6
[16] 黄仁刚. 抗菌药物对肝药酶的抑制作用及其机理. 国外医学抗生素分册, 1997;18(6):448-452
[17] Litterst CL, Mimnaugh EG, Reagan RL. Gram TE Comparison of in vitro drug metabolism by lung, liver, and kidney of several common laboratory species[J]. Drug Metab Dispos, 1975, 3(4): 259 -265.
[18] Chan DK, Mellick GD, Buchanan DD, et al. Lack of association between CYP1A1 polymorphism and Parkinson's disease in a Chinese population[J]. Neural Transm, 2002，109(1):35-39.
[19] Paine MF, Schmiedlin-Ren P, Watkins PB. Cytochrome P-450 1A1 expression in human small bowel: interindividual variation and inhibition by ketoconazole[J]. Drug Metab Dispos, 1999, 27(3):360-364.
[20] 莫启忠, Nisar A Pampor, Bernard H Shapiro. 生长激素对大鼠肾脏微粒体细胞色素P-450 4A2的影响[J]. 标记免疫分析与临床, 1996, 3(1):24-28.
[21] Ueng YF, Shyu CC, Lin YL, et al. Effects of baicalein and wogonin on drug -metabolizing enzymes in C57BL/6J mice[J]. Life Sci, 2000, 67(18):2189-2000.
[22] Biro-Sauveur B, Eeckhoutte C, Baeza E, et al. Comparison of hepatic and extrahepatic drug-metabolizing enzyme activities in rats given single or multiple challenge infections with Fasciola hepatica[J]. Int J Parasitol, 1995, 25(10):1193-2000.
[23] Nebert DW. P450 gene: structure, evolution, and regulation. Ann Rev Biochem, 1987;56:945-993
[24] Feuer G, Sosa-Lucero JC, Lumb G, et al. Failure of various drugs to induce drug-metabolizing enzymes in extrahepatic tissures of the rat. Toxicol Appl Pharmacol, 1971;19:579-589
[25] Zhang XJ, Thomas PE. Erythromycin as a specific substrate for cytochrome P450 3A isozymes and identification of a high-affinity erythromycin N-demethylase in adult female rets. Drug Metab Dispos, 1996;24(1):23-27
[26] 张均田. 《现代药理实验方法》. 第二版, 北京: 北京医科大学、北京协和医科大学联合出版社,1998;1656
[27] 徐淑云,卞如濂, 陈修. 《药理实验方法学》.第三版, 北京: 人民卫生出版社,2002;549
[28] 黄仁刚, 钱元恕. 洛美沙星和环丙沙星对小鼠肝肾的毒性作用. 中国新药杂志, 1999;8(1)：18-20
[2] Ueng YF, Shyu CC, Lin YL, et al. Effects of baicalein and wogonin on drug-metabolizing enzymes in C57BL/6J mice. Life-Sci, 2000; 67(18): 2189-2200
[3] Bohnenstengel F, Friedel G, Ritter CA, et al. Variability of cyclophosphamide uptake into human bronchial carcinoma: consequences for local bioactivation. Cancer Chemother Pharmacol. 2000; 45(1): 63-68
[4] Lieber CS. Alcohol and the liver: metabolism of alcohol and its role in hepatic and extrahepatic diseases. Mt Sinai J Med, 2000; 67(1): 84-94
[5] 周义文, 钱元恕, 盛家琦. 环丙沙星对人肝微粒体药酶活性的影响. 现代医药卫生, 2001;17（9）:668-669
[6] Valera F, De La Torre R, Segura J. Selective in vitro inhibition of hepatic oxidative metabolism by quinolones:7-ethyresorufin and caffeine as model substrates. J pharmacol, 1991;43:17
[7] Davis JD, Houston JB. Metabolism of theophylline and its inhibition by fluoroquinolones in rat hepatic microsomes. Xenobiotica, 1995;25(6):563-573
[8] Mayeaux MH, Winston GW. Amtibiotic effects on cytochromes P-450 content and mixed-function oxygenase. J Vet Pharmacol Ther, 1998;21(4):274-281
[9] 莫岚, 钱元恕, 王其南等. 培氟沙星对大鼠肝药酶的抑制作用. 新药与临床, 1995;14 (1):15-17
[10] 莫岚, 钱元恕, 王其南等. 依诺沙星对大鼠肝药酶的抑制作用. 中国抗生素杂志, 1994;19(6):459-463
[11] 黄仁刚, 钱元恕, 兰雁飞等. 洛美沙星对大鼠肝脏混合功能氧化酶的作用. 中国抗生素杂志, 1999;24(1):53-55
[12] Motomura M, Kataoka K, Takeo G, et al. Hippocampus and fromtal cortex are the potemtial mediatory sites for comvulsions induced by new quinolones and non-steroidal anti-inflammatory drugs. Int J Chin Pharmacol Ther Toxicol,1991'29(6):223-227
[13] Peng RX, Wang H, Wang YS, et al. Glutathione-related enzyme activities in human fatal adrenal, liver, and kidney. Acta Pharmacol Sinica, 1998;19(2):167-171
[14] 徐淑云,卞如濂,陈修. 《药理实验方法学》.第二版，北京:人民卫生出版社,1991;488-95
[15] 方治平,宋晓红,刘小康. 临床分离耐氟喹诺酮类金黄色葡萄球菌的gyrA基因单点突变研究. 华夏医科大学学报，2000;31（1）:24-6
[16] 黄仁刚. 抗菌药物对肝药酶的抑制作用及其机理. 国外医学抗生素分册, 1997;18(6):448-452
[17] Litterst CL, Mimnaugh EG, Reagan RL. Gram TE Comparison of in vitro drug metabolism by lung, liver, and kidney of several common laboratory species[J]. Drug Metab Dispos, 1975, 3(4): 259 -265.
[18] Chan DK, Mellick GD, Buchanan DD, et al. Lack of association between CYP1A1 polymorphism and Parkinson's disease in a Chinese population[J]. Neural Transm, 2002，109(1):35-39.
[19] Paine MF, Schmiedlin-Ren P, Watkins PB. Cytochrome P-450 1A1 expression in human small bowel: interindividual variation and inhibition by ketoconazole[J]. Drug Metab Dispos, 1999, 27(3):360-364.
[20] 莫启忠, Nisar A Pampor, Bernard H Shapiro. 生长激素对大鼠肾脏微粒体细胞色素P-450 4A2的影响[J]. 标记免疫分析与临床, 1996, 3(1):24-28.
[21] Ueng YF, Shyu CC, Lin YL, et al. Effects of baicalein and wogonin on drug -metabolizing enzymes in C57BL/6J mice[J]. Life Sci, 2000, 67(18):2189-2000.
[22] Biro-Sauveur B, Eeckhoutte C, Baeza E, et al. Comparison of hepatic and extrahepatic drug-metabolizing enzyme activities in rats given single or multiple challenge infections with Fasciola hepatica[J]. Int J Parasitol, 1995, 25(10):1193-2000.
[23] Nebert DW. P450 gene: structure, evolution, and regulation. Ann Rev Biochem, 1987;56:945-993
[24] Feuer G, Sosa-Lucero JC, Lumb G, et al. Failure of various drugs to induce drug-metabolizing enzymes in extrahepatic tissures of the rat. Toxicol Appl Pharmacol, 1971;19:579-589
[25] Zhang XJ, Thomas PE. Erythromycin as a specific substrate for cytochrome P450 3A isozymes and identification of a high-affinity erythromycin N-demethylase in adult female rets. Drug Metab Dispos, 1996;24(1):23-27
[26] 张均田. 《现代药理实验方法》. 第二版, 北京: 北京医科大学、北京协和医科大学联合出版社,1998;1656
[27] 徐淑云,卞如濂, 陈修. 《药理实验方法学》.第三版, 北京: 人民卫生出版社,2002;549
[28] 黄仁刚, 钱元恕. 洛美沙星和环丙沙星对小鼠肝肾的毒性作用. 中国新药杂志, 1999;8(1)：18-20