中国中药杂志

2009, v.34(03) 307-311

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18β-甘草酸和18α-甘草酸对大鼠原代肝细胞CYP3A酶表达的影响
Effects of 18β-glycyrrhizic acid and 18α-glycyrrhizic acid on mRNA and protein expression of cytochrome P450 3A in cultured rat primary hepatocyte

王宇光;杨明会;马增春;梁乾德;谭洪玲;肖成荣;高月;
WANG Yuguang1,YANG Minghui2,MA Zengchun1,LIANG Qiande1,TAN Hongling1,XIAO Chengrong1,GAO Yue1(1.Beijing Institute of Radiation Medicine,Beijing 100850,China; 2.Institute of Traditional Chinese Medicine,General Hospital of PLA,Beijing 100853,China)

摘要(Abstract):

目的:研究甘草中的2种成分18β-甘草酸和18α-甘草酸对原代培养大鼠肝细胞中细胞色素P450 3A(CYP3A)在mRNA及蛋白水平表达的影响,并探讨其剂量-效应关系。方法:原位两步胶原酶灌流法分离大鼠肝细胞并采用"胶原蛋白凝胶三明治"培养原代肝细胞,无血清条件下培养细胞,并用不同剂量的18β-甘草酸和18α-甘草酸处理细胞,RT-PCR法测定CYP3A mRNA表达水平,Western-blot法测定CYP3A蛋白表达。结果:实验条件下对照组、18β-甘草酸、18α-甘草酸处理组CYP3A基因及蛋白均可被检测,18β-甘草酸浓度依赖性诱导CYP3A mRNA(25100μmol.L-1)及蛋白表达(50100μmol.L-1)及蛋白表达(50400μmol.L-1),而18α-甘草酸浓度依赖性抑制CYP3A mRNA(25400μmol.L-1),而18α-甘草酸浓度依赖性抑制CYP3A mRNA(25100μmol.L-1)及蛋白表达(25100μmol.L-1)及蛋白表达(25100μmol.L-1)表达。结论:18β-甘草酸和18α-甘草酸在转录水平上分别上调和下调大鼠肝细胞CYP3A的表达。
Objective:To study the effects of 18β-glycyrrhizic acid and 18α-glycyrrhizic acid on mRNA and protein expression of cytochrome P450 3A in cultured rat primary hepatocyte.Method:Rat heaptocyte were isolated by two-step in situ collagenase perfusion method;the hepatocytes were seeded in dishes coated with type I rat tail collagen,culture medium was added and changed daily after gelation,the effect of 18β-glycyrrhizic acid and 18α-glycyrrhizic acid on the expression of CYP3A was determined by reverse transcriptase-polymerase chain reaction(RT-PCR) and western blot respectively.Result:Treatment with 18β-glycyrrhizic acid of primary rat hepatocytes resulted in marked up-regulationof CYP3A1expression at both mRNA and protein levels in a concentration dependent manner,while exposure to 18α-glycyrrhizic acid of primary rat hepatocytes resulted in marked down-regulation of CYP3A1expression at both mRNA and protein levels in a concentration dependent manmer.Conclusion:18β-glycyrrhizic acid and 18α-glycyrrhizic acid up-regulate and down-regulate CYP3A1 expression at the transcriptive levels.

关键词(KeyWords): 18β-甘草酸;18α-甘草酸;CYP3A;原代肝细胞
18β-glycyrrhizic acid;18α-glycyrrhizic acid;CYP3A

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(30070936,30701112)

作者(Authors): 王宇光;杨明会;马增春;梁乾德;谭洪玲;肖成荣;高月;
WANG Yuguang1,YANG Minghui2,MA Zengchun1,LIANG Qiande1,TAN Hongling1,XIAO Chengrong1,GAO Yue1(1.Beijing Institute of Radiation Medicine,Beijing 100850,China; 2.Institute of Traditional Chinese Medicine,General Hospital of PLA,Beijing 100853,China)

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