基于“异类相制”理论的雷公藤复方配伍对CYP450酶系的影响
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摘要
目的:以"异类相制"理论为指导,观察雷公藤不同中药复方配伍后对雷公藤肝脏毒性的影响,并探讨其机制。方法:将SD大鼠随机分为8组,分别为:空白组、雷公藤单药组、清络通痹复方组、雷公藤多苷片组、雷公藤+生地黄组、雷公藤+三七组、雷公藤+青风藤组、雷公藤+僵蚕组,灌胃给药30d。以生化和病理检查综合评估肝毒性,并采用"Cocktail"底物探针法评价雷公藤的不同配伍对大鼠肝微粒体体外温孵体系中细胞色素P450酶(CYP450)各亚型的影响。结果:雷公藤单药组、雷公藤多苷片组、雷公藤+青风藤组、雷公藤+僵蚕组与空白组比较可见多项生化指标明显差异,服用清络通痹复方、配伍三七或生地黄可较单药组降低ALT、AST指标(P<0.01,P<0.05)。病理检查回示,雷公藤和不同配方组合应用后,肝细胞损伤程度减轻。各用药组较空白组CYP3A、CYP2C9、CYP2C19的特异性底物终浓度增高,其羟基化代谢产物终浓度下降(P<0.05,P<0.01),提示服用雷公藤可导致肝内CPY3A、CYP2C9,CPY2C19酶活性的下降。和单药组对比,雷公藤经配伍后均可改善单药对CYP3A和CYP2C19的抑制。其中,三七对改善雷公藤单药对CYP3A的抑制作用比较明显(P<0.05),而生地黄对改善雷公藤单药对CYP2C19的抑制作用比较明显(P<0.05)。结论:清络通痹复方配伍可明显减轻雷公藤的肝毒性。经一一拆方与雷公藤配伍后,发现雷公藤与生地黄和三七配伍后可减轻其肝毒性,其减毒机制可能与生地黄和三七缓解了雷公藤对CYP3A和CYP2C19的抑制相关。
Objective: To observe the effects of compound prescription of tripterygium wilfordii on reducing liver toxicity of tripterygium wilfordii based on theory of ‘heterogeneous restriction'. Methods: SD rats were randomly divided into 8 groups as control group, tripterygium wilfordii group, compound prescription of tripterygium wilfordii group, Qingluo Tongbi group, glucosidorum tripterygll totorum tablets group, tripterygium wilfordii+unprocessed rehmannia root group, tripterygium wilfordii+orientvine vine group, and tripterygium wilfordii+stiff silkworm group. After treating for 30 days, the liver toxicity was assessed synthetically by using biochemical and pathological examination. The influence of different concerted applications of tripterygium wilfordii on CYP450 enzyme family of rat liver microsomes in vitro incubation system was assessed by using Cocktail substrate probe method. Results: There were significant differences in a number of biochemical indicators among control group, tripterygium wilfordii group, glucosidorum tripterygll totorum tablets group, tripterygium wilfordii+orientvine vine group, and tripterygium wilfordii+stiff silkworm group. Qingluo Tongbi Formula, and tripterygium wilfordii+unprocessed rehmannia root or sanqi could decrease the level of ALT, AST(P<0.01, P<0.05), and the effects of Qingluo Tongbi Formulais the most significant. Compared with tripterygium wilfordii group, tripterygium wilfordii+sanqi could decrease the level of AST significantly(P<0.05, P<0.01). Histopathological examination revealed that combination use of tripterygium wilfordii and different concerted applications could reduce liver cell damage degree. The specific substrate concentration of CYP3 A, CYP2C9, and CYP2C19 of drug group was higher than that of control group, and the final concentration of hydroxylated metabolites of drug groups decreased(P<0.05, P<0.01), which suggesting that taking tripterygium wilfordii can lead to activity decreasing of CPY3 A, CYP2C9, and CPY2C19 in liver. Compared with single drug group, concerted application of tripterygium wilfordii could improve the inhibiting effect on CYP3 A and CYP2C19 from single drug. Sanqi could significantly improve the inhibiting effect on CYP3 A from tripterygium wilfordii(P<0.05), and unprocessed rehmannia root could significantly improve the inhibiting effect on CYP2C19 from tripterygium wilfordii(P<0.05). Conclusion: Qingluo Tongbi Formula could significantly reduce the liver toxicity of tripterygium wilfordii. Concerted application of tripterygium wilfordii and form drugs of Qingluo Tongbi Formula prompted that concerted application of tripterygium wilfordii and sanqi or unprocessed rehmannia root could reduce the liver toxicity of tripterygium wilfordii, and the mechanism might relate with the relaxation effect of sanqi and unprocessed rehmannia root on inhibiting CYP3 A and CYP2C19 caused by tripterygium wilfordii.
Objective: To observe the effects of compound prescription of tripterygium wilfordii on reducing liver toxicity of tripterygium wilfordii based on theory of ‘heterogeneous restriction'. Methods: SD rats were randomly divided into 8 groups as control group, tripterygium wilfordii group, compound prescription of tripterygium wilfordii group, Qingluo Tongbi group, glucosidorum tripterygll totorum tablets group, tripterygium wilfordii+unprocessed rehmannia root group, tripterygium wilfordii+orientvine vine group, and tripterygium wilfordii+stiff silkworm group. After treating for 30 days, the liver toxicity was assessed synthetically by using biochemical and pathological examination. The influence of different concerted applications of tripterygium wilfordii on CYP450 enzyme family of rat liver microsomes in vitro incubation system was assessed by using Cocktail substrate probe method. Results: There were significant differences in a number of biochemical indicators among control group, tripterygium wilfordii group, glucosidorum tripterygll totorum tablets group, tripterygium wilfordii+orientvine vine group, and tripterygium wilfordii+stiff silkworm group. Qingluo Tongbi Formula, and tripterygium wilfordii+unprocessed rehmannia root or sanqi could decrease the level of ALT, AST(P<0.01, P<0.05), and the effects of Qingluo Tongbi Formulais the most significant. Compared with tripterygium wilfordii group, tripterygium wilfordii+sanqi could decrease the level of AST significantly(P<0.05, P<0.01). Histopathological examination revealed that combination use of tripterygium wilfordii and different concerted applications could reduce liver cell damage degree. The specific substrate concentration of CYP3 A, CYP2C9, and CYP2C19 of drug group was higher than that of control group, and the final concentration of hydroxylated metabolites of drug groups decreased(P<0.05, P<0.01), which suggesting that taking tripterygium wilfordii can lead to activity decreasing of CPY3 A, CYP2C9, and CPY2C19 in liver. Compared with single drug group, concerted application of tripterygium wilfordii could improve the inhibiting effect on CYP3 A and CYP2C19 from single drug. Sanqi could significantly improve the inhibiting effect on CYP3 A from tripterygium wilfordii(P<0.05), and unprocessed rehmannia root could significantly improve the inhibiting effect on CYP2C19 from tripterygium wilfordii(P<0.05). Conclusion: Qingluo Tongbi Formula could significantly reduce the liver toxicity of tripterygium wilfordii. Concerted application of tripterygium wilfordii and form drugs of Qingluo Tongbi Formula prompted that concerted application of tripterygium wilfordii and sanqi or unprocessed rehmannia root could reduce the liver toxicity of tripterygium wilfordii, and the mechanism might relate with the relaxation effect of sanqi and unprocessed rehmannia root on inhibiting CYP3 A and CYP2C19 caused by tripterygium wilfordii.
引文
[1]周学平,周玲玲,王旭.基于“异类相制”理论探讨中药复方的配伍减毒作用.中医杂志,2013,54(4):271-273
[2]张静,江莹,王芳,等.基于“异类相制”理论探讨雷公藤肝毒性配伍减毒的作用.中草药,2014,45(18):2711-2715
[3]刘为萍,刘素香,唐慧珠,等.雷公藤研究新进展.中草药,2010,41(7):1215-1218
[4]李桓,周学平,陆艳.雷公藤“异类相制”配伍减毒作用探讨.中医杂志,2015,56(01):10-13
[5]周玲玲,周学平,梁卫,等.清络通痹颗粒对胶原关节炎大鼠的保护作用及机制研究.中药药理与临床,2011,27(2):109-112
[6]周玲玲,柳璋璞,梁晓雯,等.清络通痹颗粒对滑膜成纤维细胞与单核细胞共培养诱生的破骨样细胞的影响.中国免疫学杂志,2011,27(11):988-992
[7]周聪,周玲玲,周学平.中药配伍雷公藤减肝毒性机制研究.中华中医药杂志,2014,29(4):1167-1169
[8]赵莉,周学平.中药配伍减轻雷公藤生殖毒性的研究进展.中华中医药杂志,2015,30(2):482-484
[9]候丛颂,杨志宏,孙晓波.“cocktail”探针药物法及其在研究中药对细胞色素P450影响中的应用进展.中国药理学与毒理学杂志,2013,27(3):445-450
[10]Dany Spaggiari,Laurent Geiser,Youssef Daali.A cocktail approach for assessing the in vitro activity of human cytochrome P450s:An overview of current methodologies.Journal of Pharmaceutical and Biomedical Analysis,2014,101(3):221-237
[11]Youdim K A,Saunders K C.A review of LC-MS techniques and high-throughput approaches used to investigate drug metabolism by cytochrome P450s.Journal of Chromatography B,2010,878(5):1326-1336
[12]魏伟,吴希美,李元建.药理实验方法学.北京:人民卫生出版社,2010:188-189
[13]陈五海,周学平.自拟清络通痹方治疗强直性脊柱炎湿热痹阻证的临床观察.云南中医中药杂志,2011,32(11):40-42
[14]Evans W E,Relling M V.Pharmacogenomics:translating functional genomics into rational therapeutics,Science,1999,286(10):487-491
[15]Gu J,Weng Y,Zhang Q Y.Liver-specific deletion of the NADPH-cytochrome P450 reductase gene:impact on plasma cholesterol homeostasis and the function and regulation of microsomal cytochrome P450 and heme oxygenase.Biol Chem,2003,278(7):25895-25901
[16]Xiang X,Likun G,Xinming Q,et al.Knockout of hepatic P450reductase aggravates triptolide-induced toxicity.Toxicology Letters,2011,205(8):47-54
[17]Li L,Zhen Z J,Jing L,et al.Sex differences in subacute toxicity and hepatic microsomal metabolism of triptolide in rats.Toxicology2010,271(4):57-63
[18]叶永山,汤明杰,张旗,等.生地黄药效物质基础研究进展.中华中医药学会中药化学分会第八届学术年会论文集,2013:8
[19]高贝贝,折改梅.三七化学成分研究进展.中国商品学会.第二届全国中药商品学术大会论文集,2010:8
[20]陆艳,谢彤,李桓,等.从中药复方配伍减毒的物质基础探讨“异类相制”理论的研究思路.南京中医药大学学报,2016,32(1):97-100
[2]张静,江莹,王芳,等.基于“异类相制”理论探讨雷公藤肝毒性配伍减毒的作用.中草药,2014,45(18):2711-2715
[3]刘为萍,刘素香,唐慧珠,等.雷公藤研究新进展.中草药,2010,41(7):1215-1218
[4]李桓,周学平,陆艳.雷公藤“异类相制”配伍减毒作用探讨.中医杂志,2015,56(01):10-13
[5]周玲玲,周学平,梁卫,等.清络通痹颗粒对胶原关节炎大鼠的保护作用及机制研究.中药药理与临床,2011,27(2):109-112
[6]周玲玲,柳璋璞,梁晓雯,等.清络通痹颗粒对滑膜成纤维细胞与单核细胞共培养诱生的破骨样细胞的影响.中国免疫学杂志,2011,27(11):988-992
[7]周聪,周玲玲,周学平.中药配伍雷公藤减肝毒性机制研究.中华中医药杂志,2014,29(4):1167-1169
[8]赵莉,周学平.中药配伍减轻雷公藤生殖毒性的研究进展.中华中医药杂志,2015,30(2):482-484
[9]候丛颂,杨志宏,孙晓波.“cocktail”探针药物法及其在研究中药对细胞色素P450影响中的应用进展.中国药理学与毒理学杂志,2013,27(3):445-450
[10]Dany Spaggiari,Laurent Geiser,Youssef Daali.A cocktail approach for assessing the in vitro activity of human cytochrome P450s:An overview of current methodologies.Journal of Pharmaceutical and Biomedical Analysis,2014,101(3):221-237
[11]Youdim K A,Saunders K C.A review of LC-MS techniques and high-throughput approaches used to investigate drug metabolism by cytochrome P450s.Journal of Chromatography B,2010,878(5):1326-1336
[12]魏伟,吴希美,李元建.药理实验方法学.北京:人民卫生出版社,2010:188-189
[13]陈五海,周学平.自拟清络通痹方治疗强直性脊柱炎湿热痹阻证的临床观察.云南中医中药杂志,2011,32(11):40-42
[14]Evans W E,Relling M V.Pharmacogenomics:translating functional genomics into rational therapeutics,Science,1999,286(10):487-491
[15]Gu J,Weng Y,Zhang Q Y.Liver-specific deletion of the NADPH-cytochrome P450 reductase gene:impact on plasma cholesterol homeostasis and the function and regulation of microsomal cytochrome P450 and heme oxygenase.Biol Chem,2003,278(7):25895-25901
[16]Xiang X,Likun G,Xinming Q,et al.Knockout of hepatic P450reductase aggravates triptolide-induced toxicity.Toxicology Letters,2011,205(8):47-54
[17]Li L,Zhen Z J,Jing L,et al.Sex differences in subacute toxicity and hepatic microsomal metabolism of triptolide in rats.Toxicology2010,271(4):57-63
[18]叶永山,汤明杰,张旗,等.生地黄药效物质基础研究进展.中华中医药学会中药化学分会第八届学术年会论文集,2013:8
[19]高贝贝,折改梅.三七化学成分研究进展.中国商品学会.第二届全国中药商品学术大会论文集,2010:8
[20]陆艳,谢彤,李桓,等.从中药复方配伍减毒的物质基础探讨“异类相制”理论的研究思路.南京中医药大学学报,2016,32(1):97-100