摘要
目的研究异鼠李素对肝脏6种CYPs的体外抑制作用,以及对大鼠原代肝细胞的毒性作用。方法采用人肝微粒体(HLMs)体外温孵法研究异鼠李素对6种细胞色素P450酶(CYPs)——CYP2C19、CYP2D6、CYP3A4、CYP2E1、CYP1A2和CYP2C9的体外抑制作用;使用HPLC-MS/MS法检测异鼠李素和HLMs共同孵育后的代谢产物;利用体外培养的低CYPs活性的大鼠原代肝细胞,考察不同剂量异鼠李素对细胞培养液中乳酸脱氢酶(LDH)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)的影响。结果 50μmol/L的异鼠李素对CYP2E1和CYP1A2有一定的抑制作用,抑制率分别为59.48%和39.91%;异鼠李素和HLMs共同孵育后,产生去甲基化代谢产物3,3’,4’,5,7-五羟基黄酮,转化为极性和水溶性较高的代谢物;30、100、300μmol/L的异鼠李素会使大鼠原代肝细胞培养液中的ALT和LDH显著上升(P<0.01),100、300μmol/L异鼠李素使AST显著上升(P<0.05、0.01),呈浓度相关性。结论异鼠李素在体外主要经HLMs代谢,同时对CYP2E1和CYP1A2有一定的抑制作用,可能会使CYP2E1和CYP1A2的底物药物在体内的浓度产生变化,导致一系列药物的相互作用;大量使用异鼠李素可能会造成一定程度的肝细胞损伤,且呈现浓度相关性。临床应用应合理设置剂量,并注意潜在的药物之间的相互作用。
Objective To study the inhibitory effects ofisorhamnetin on six kinds of CYPs of liver in vitro,and the toxic effect on rat hepatocytes Methods This report uses warm incubation of human liver microsomes in vitro to investigate the inhibition of isorhamnetin on 6 kinds of CYPs （CYP2C19,CYP2D6,CYP3A4,CYP2E1,CYP1A2 and CYP2C9）,and using HPLC-MS/MS to detect product of metabolism as well as analysing of the pathways of metabolic.At the same time,using rat primary hepatocytes which has low CYPs activity in vitro to explore whether the use of isorhamnetin will cause effects on the ALT,AST and LDH of hepatocytes.Results Isorhamnetin has inhibition effects on CYP2E1 and CYP1A2,the inhibition rate were 59.48% and 39.91%,respectively.Methylated metabolite is produced after incubating of isorhamnetin and HLMs.The isorhmnetin becomes high polarity and water solubility metabolite 3,3＆#39;,4＆#39;,5,7-hydroxyflavone.Isorhamnetin of 30,100 and 300 μmol/L cause a significant rise of ALT and LDH in primary cultured rat hepatocytes cultured （P 〈 0.01）.isorharnnetin of 100 μmol/L cause a rise of AST in primary cultured rat hepatocytes cultured （P 〈 0.05） and 300 μmol/L cause a significant rise （P 〈 0.01）.It was a dose-dependent manner.Conclusion Isorhamnetin in vitro mainly metabolized by HLMs,and at the same time have a certain inhibitory effect on CYP2E1 and CYP1A2,which may cause the drugs which are metabolized by CYP2E1 and CYP1A2 in vivo accumulation that lead to a series of drug interactions.The results also indicate that heavy use of isorhamnetin cause some adverse effects on hepatocytes,and it was a dose-dependent manner.Individuals need to pay attention to the dose ofisorhamnetin and the potential drug interactions.
出处
《药物评价研究》
CAS
2017年第5期627-632,共6页
Drug Evaluation Research
基金
国家自然科学基金面上项目(81373890)
教育部"创新团队发展计划"(IRT_14R41)
