菌群失调大鼠体内PEG400对黄芩苷和黄芩素药代动力学的影响
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摘要
建立UPLC-MS/MS研究在正常大鼠和肠道菌群失调大鼠体内,聚乙二醇400(PEG400)对黄芩苷、黄芩素在正常大鼠和肠道菌群失调大鼠体内的药代动力学的影响。以染料木素为内标,血浆样品经乙酸乙酯沉淀蛋白,UPLC-MS/MS测定,方法经专属性、线性范围、准确度、精密度、稳定性等确证适合用于血浆中黄芩苷的测定。采用盐酸林可霉素(5 g·kg-1·d-1)造模1周,诱导菌群失调大鼠模型。正常大鼠和肠道菌群失调大鼠分别灌胃黄芩苷、黄芩苷+PEG400、黄芩素和黄芩素+PEG400后不同时间点取血浆,UPLC-MS/MS测定给药后血浆中黄芩苷的浓度,采用DAS 3. 2. 2软件计算药动学参数,绘制血药浓度-时间曲线图。结果显示盐酸林可霉素诱导的菌群失调大鼠粪便中β-葡萄糖醛酸苷酶活性和菌落数显著降低。肠道菌群失调大鼠黄芩苷+PEG400组的Cmax和AUC0-t显著低于正常大鼠黄芩苷+PEG400组。菌群失调大鼠黄芩苷组和黄芩苷+PEG400组的Cmax和AUC0-t无显著性差异。正常大鼠黄芩苷组的Cmax和AUC0-t显著高于肠道菌群失调大鼠黄芩苷组和黄芩苷+PEG400组。正常大鼠黄芩素+PEG400组的Cmax和AUC0-t与黄芩素组无显著性差异。菌群失调大鼠黄芩素组的Cmax和AUC0-t低于正常组但显著高于黄芩素+PEG400组。结果表明,PEG400能够增加正常大鼠体内黄芩苷的吸收,菌群失调大鼠体内则无效,不影响黄芩素在大鼠体内的吸收。
The study aimed to establish an UPLC-MS/MS method for the determination of baicalin in rat plasma,in order to study the effect of PEG400 on pharmacokinetics of baicalin and baicalein in normal and gut microbiotadysbiosis rats. Plasma was precipitated with ethyl acetate and determined by UPLC-MS/MS method,with genistein as an internal standard. In terms of specificity,linearity,range,accuracy,precision and stability,the method was suitable for the determination of baicalin in plasma. The gut microbiotadysbiosis rat model was induced through the oral administration with lincomycin hydrochloride(5 g·kg-1·d-1) for one week. Samples of plasma of rats were obtained at different time points,after the rats were administrated with baicalin,baicalin and PEG400. Baicalin in rats were detected by UPLC-MS/MS method,and pharmacokinetic parameters were calculated by DAS 3. 2. 2 software. The results showed that the β-glucosidase activity and the number of colonies in the feces of gut microbiotadysbiosis rats induced by lincomycin hydrochloride were significantly reduced. The Cmaxand AUC0-tof the baicalinand PEG400 group in the intestinal flora were significantly lower than those in the normal rat baicalin and PEG400 group. There was no significant difference in Cmaxand AUC0-tbetween the baicalin group and the baicalin+PEG400 group of gut microbiotadysbiosis rats. The Cmaxand AUC0-tof the normal rats baicalin group were significantly higher than those of the gut microbiotadysbiosis rats baicalin group and the baicalin + PEG400 group. There was no significant difference in Cmaxand AUC0-tbetween the normal rat baicalein and PEG400 group and the baicalein group. The Cmaxand AUC0-tof the baicalein group in the gut microbiotadysbiosis rats were lower than those in the normal baicalein group,but significantly higher than those in the baicalein and PEG400 group. PEG400 could increase the absorption of baicalin in normal rats,but is ineffective in gut microbiotadysbiosis rats,with no impact on the absorption of baicalein in rats.
The study aimed to establish an UPLC-MS/MS method for the determination of baicalin in rat plasma,in order to study the effect of PEG400 on pharmacokinetics of baicalin and baicalein in normal and gut microbiotadysbiosis rats. Plasma was precipitated with ethyl acetate and determined by UPLC-MS/MS method,with genistein as an internal standard. In terms of specificity,linearity,range,accuracy,precision and stability,the method was suitable for the determination of baicalin in plasma. The gut microbiotadysbiosis rat model was induced through the oral administration with lincomycin hydrochloride(5 g·kg-1·d-1) for one week. Samples of plasma of rats were obtained at different time points,after the rats were administrated with baicalin,baicalin and PEG400. Baicalin in rats were detected by UPLC-MS/MS method,and pharmacokinetic parameters were calculated by DAS 3. 2. 2 software. The results showed that the β-glucosidase activity and the number of colonies in the feces of gut microbiotadysbiosis rats induced by lincomycin hydrochloride were significantly reduced. The Cmaxand AUC0-tof the baicalinand PEG400 group in the intestinal flora were significantly lower than those in the normal rat baicalin and PEG400 group. There was no significant difference in Cmaxand AUC0-tbetween the baicalin group and the baicalin+PEG400 group of gut microbiotadysbiosis rats. The Cmaxand AUC0-tof the normal rats baicalin group were significantly higher than those of the gut microbiotadysbiosis rats baicalin group and the baicalin + PEG400 group. There was no significant difference in Cmaxand AUC0-tbetween the normal rat baicalein and PEG400 group and the baicalein group. The Cmaxand AUC0-tof the baicalein group in the gut microbiotadysbiosis rats were lower than those in the normal baicalein group,but significantly higher than those in the baicalein and PEG400 group. PEG400 could increase the absorption of baicalin in normal rats,but is ineffective in gut microbiotadysbiosis rats,with no impact on the absorption of baicalein in rats.
引文
[1]冯友根,邹晓华.国内黄芩苷药理研究和临床应用概述[J].中国医药情报,2004,10(2):35.
[2] Kyun Ha Kim,Young-Don Park,Heejin Park,et al. Synthesis and biological evaluation of a novel baicalein glycoside as an anti-inflammatory agent[J]. Eur J Pharm Sci,2014,744(11):147.
[3] Paul Wai Kei,Tsang,Chau Kwok-Yan,et al. Baicalein exhibits inhibitory effect on the energy-dependent efflux pump activity in nonalbicans Candida fungi[J]. Chemotherapy, 2015, 27(161):221.
[4] Gao L,Fang J S,Bai X Y,et al. In silico target fishing for the potential targets and molecular mechanisms of baicalein as an antiparkinsonian agent:discovery of the protective effects on NMDA receptor-mediated neurotoxicity[J]. Chem Biol Drug Des,2013,81(6):675.
[5] Middleton E J,Kandaswami C,Theoharides T C. The effects of plant flavonoids onmammalian cells:implications for inflammation,heart disease and cancer[J]. Pharmacol Rev,2000,52(4):673.
[6] Ricc E C. Flavonoids antioxidants[J]. Curr Med Chem,2001,8(7):797.
[7] Kubo M,Kimura Y,Odani T,et al. Studies on Scutellaria Radix part 2:the antibacterial substance[J]. Planta Med,1981,43(2):194.
[8]车庆明,杨琳,陈颖,等.不同剂量黄芩素在大鼠体内的药动学差异[J].中国新药杂志,2007,16(8):604.
[9]何秀琼,裴利霞,王一涛,等.高效液相色谱法研究大鼠灌胃黄芩素的药动学[J].中国医院药学杂志,2010,30(22):1901.
[10]周红潮,杜锐.黄芩苷药代动力学研究进展[J].中国中药杂志,2018,43(4):684.
[11]郭晓宇,杨琳,陈颖,等.黄芩素与黄芩苷大鼠体内药动学比较研究[J].中国药学杂志,2008,43(7):524.
[12]周乐,赵晓莉,狄留庆,等.黄酮类化合物口服吸收与代谢特征及其规律分析[J].中草药,2013,44(16):2313.
[13] Graf B,Aeho C,Dolnikowkig G,et al. Rat gastrointinal tissues metabolize quercetin[J]. J Nutr,2006,136(1):39.
[14] Van D H,Boersmam G,Vervoort J,et al. Identification of 14quercetin phaseⅡmono-and mixed conjugates and their formation by ratand human phaseⅡin vitro model systems[J]. Chem Res Toxicol,2004,17(11):1520.
[15] Gradolatto A,Canyvenglavier M,Basly J P,et al. Metabolism of apigenin by liver phaseⅠand phaseⅡenzymes and by isolated perfused rat liver[J]. J Drug Metab Dispos,2004,32(1):58.
[16]毛凤斐,屠锡德,朱家璧,等.黄芩甙在人体内吸收的研究[J].南京药学院学报,2012,5(1):70.
[17] Zhang L,Lin G,Zuo Z. Involvement of UDP-glucuronosyltransferases in the extensive liver and intestinal first-pass metabolism of flavonoid baicalein[J]. Pharm Res,2012,24(1):81.
[18] Zhang L,Lin G,Kovács B,et al. Mechanistic study on the intestinal absorption and disposition of baicalein[J]. Eur J Pharm Sci,2013,31(3/4):221.
[19]黄建耿.药用辅料抑制肠道P-糖蛋白药泵作用研究[D].武汉:华中科技大学,2008.
[20] Jianmei D,Hai M,Christopher R,et al. Understanding the degradation pathway of a poorly water-soluble drug formulated in PEG-400[J].Pharmaceut Biomed,2007,88(7):1638.
[21] Teofilo V,Bruno S,Paulo C,et al. Solid dispersions as strategy to improve oral bioavailability of poor water soluble drugs[J].Drug Discov Today,2007,23(12):1068.
[22]孟小夏,舒阳,张敏,等.聚乙二醇400对黄芩苷在大鼠体内药动学的影响[J].华西药学杂志,2017,32(2):132.
[23]许海舰,刘一鑫,刘喜纲,等.黄芩苷镁盐和黄芩苷的肠吸收动力学和药代动力学比较[J].中国实验方剂学杂志,2018,24(4):78.
[24] Fishman W H,Kato K,Anstiss C L,et al. Human serumβ-glucuronidase:its measurement and some of its properties[J]. Clin Chim Acta,1967,15:435.
[25] Sullivan A,Barkholt L,Nord C E. Lactobacillusacidophilus,Bifidobacteriumlactis and Lactobacillus F19 prevent antibiotic-associated econological disturbance of bacteroides fragilis in the intestine[J]. Antimicrob Chemother,2003,52(2):308.
[2] Kyun Ha Kim,Young-Don Park,Heejin Park,et al. Synthesis and biological evaluation of a novel baicalein glycoside as an anti-inflammatory agent[J]. Eur J Pharm Sci,2014,744(11):147.
[3] Paul Wai Kei,Tsang,Chau Kwok-Yan,et al. Baicalein exhibits inhibitory effect on the energy-dependent efflux pump activity in nonalbicans Candida fungi[J]. Chemotherapy, 2015, 27(161):221.
[4] Gao L,Fang J S,Bai X Y,et al. In silico target fishing for the potential targets and molecular mechanisms of baicalein as an antiparkinsonian agent:discovery of the protective effects on NMDA receptor-mediated neurotoxicity[J]. Chem Biol Drug Des,2013,81(6):675.
[5] Middleton E J,Kandaswami C,Theoharides T C. The effects of plant flavonoids onmammalian cells:implications for inflammation,heart disease and cancer[J]. Pharmacol Rev,2000,52(4):673.
[6] Ricc E C. Flavonoids antioxidants[J]. Curr Med Chem,2001,8(7):797.
[7] Kubo M,Kimura Y,Odani T,et al. Studies on Scutellaria Radix part 2:the antibacterial substance[J]. Planta Med,1981,43(2):194.
[8]车庆明,杨琳,陈颖,等.不同剂量黄芩素在大鼠体内的药动学差异[J].中国新药杂志,2007,16(8):604.
[9]何秀琼,裴利霞,王一涛,等.高效液相色谱法研究大鼠灌胃黄芩素的药动学[J].中国医院药学杂志,2010,30(22):1901.
[10]周红潮,杜锐.黄芩苷药代动力学研究进展[J].中国中药杂志,2018,43(4):684.
[11]郭晓宇,杨琳,陈颖,等.黄芩素与黄芩苷大鼠体内药动学比较研究[J].中国药学杂志,2008,43(7):524.
[12]周乐,赵晓莉,狄留庆,等.黄酮类化合物口服吸收与代谢特征及其规律分析[J].中草药,2013,44(16):2313.
[13] Graf B,Aeho C,Dolnikowkig G,et al. Rat gastrointinal tissues metabolize quercetin[J]. J Nutr,2006,136(1):39.
[14] Van D H,Boersmam G,Vervoort J,et al. Identification of 14quercetin phaseⅡmono-and mixed conjugates and their formation by ratand human phaseⅡin vitro model systems[J]. Chem Res Toxicol,2004,17(11):1520.
[15] Gradolatto A,Canyvenglavier M,Basly J P,et al. Metabolism of apigenin by liver phaseⅠand phaseⅡenzymes and by isolated perfused rat liver[J]. J Drug Metab Dispos,2004,32(1):58.
[16]毛凤斐,屠锡德,朱家璧,等.黄芩甙在人体内吸收的研究[J].南京药学院学报,2012,5(1):70.
[17] Zhang L,Lin G,Zuo Z. Involvement of UDP-glucuronosyltransferases in the extensive liver and intestinal first-pass metabolism of flavonoid baicalein[J]. Pharm Res,2012,24(1):81.
[18] Zhang L,Lin G,Kovács B,et al. Mechanistic study on the intestinal absorption and disposition of baicalein[J]. Eur J Pharm Sci,2013,31(3/4):221.
[19]黄建耿.药用辅料抑制肠道P-糖蛋白药泵作用研究[D].武汉:华中科技大学,2008.
[20] Jianmei D,Hai M,Christopher R,et al. Understanding the degradation pathway of a poorly water-soluble drug formulated in PEG-400[J].Pharmaceut Biomed,2007,88(7):1638.
[21] Teofilo V,Bruno S,Paulo C,et al. Solid dispersions as strategy to improve oral bioavailability of poor water soluble drugs[J].Drug Discov Today,2007,23(12):1068.
[22]孟小夏,舒阳,张敏,等.聚乙二醇400对黄芩苷在大鼠体内药动学的影响[J].华西药学杂志,2017,32(2):132.
[23]许海舰,刘一鑫,刘喜纲,等.黄芩苷镁盐和黄芩苷的肠吸收动力学和药代动力学比较[J].中国实验方剂学杂志,2018,24(4):78.
[24] Fishman W H,Kato K,Anstiss C L,et al. Human serumβ-glucuronidase:its measurement and some of its properties[J]. Clin Chim Acta,1967,15:435.
[25] Sullivan A,Barkholt L,Nord C E. Lactobacillusacidophilus,Bifidobacteriumlactis and Lactobacillus F19 prevent antibiotic-associated econological disturbance of bacteroides fragilis in the intestine[J]. Antimicrob Chemother,2003,52(2):308.