人参、甘草与附子配伍调控CYP450系统减轻肝毒性研究

Study on regulation of CYP450 enzyme system to reduce liver toxicity through the compatibility of radix aconiti lateralis praeparata with Panax ginseng C. A. Mey and Glycyrrhiza uralensis Fisch

目的:考察人参、甘草与附子配伍后对正常大鼠肝脏细胞色素P450 (cytochrome P450,CYP450) mRNA转录水平及蛋白翻译水平的影响。方法:雄性SD大鼠,随机分为空白对照组、诱导剂组、抑制剂组、人参组、甘草组、附子组、人参+附子组、甘草+附子组和人参+甘草+附子组,每组6只大鼠。检测大鼠血清总胆汁酸(TBA),碱性磷酸酶(ALP),谷丙转氨酶(ALT)和谷草转氨酶(AST)含量变化,观察大鼠肝脏组织的病理学改变,并观察各组CYP1A2和CYP3A1的mRNA和蛋白表达水平的变化。结果:与空白对照组相比,附子组中大鼠血清TBA,ALP,ALT和AST的含量升高,配伍后TBA,ALP,ALT和AST的含量较附子组降低;同时配伍后可减轻附子产生的肝脏损伤作用;附子组对CYP1A2和CYP3A1mRNA的表达具有抑制作用,与人参、甘草配伍后可上调该亚型的表达;翻译水平与转录水平一致。结论:人参、甘草与附子配伍后可上调CYP450酶的表达,减少乌头碱在体内蓄积时间,起到减毒的作用,CYP450上调增量大小顺序为人参+甘草+附子组>甘草+附子组>人参+附子组。

Objective:To explore the effect of Panax ginseng C.A.Mey(RS)and Glycyrrhiza uralensis Fisch.(GC)combined with radix aconiti lateralis praeparata(FZ)on mRNA and protein expression of cytochrome P450(CYP450)enzymes in rat liver.Methods:Male Sprague-Dawley rats were randomly divided into negative control group,inducer group,inhibitor group,FZ group,RS group,GC group,RS+FZ group,GC+FZ group and RS+GC+FZ group w让h 6 rats in each group.In each group,the serum concentrations of TBA,ALP,ALT and AST were respectively measured and the pathologic changes was detected using HE staining in the liver tissue of the rats.The expression levels of mRNA and total protein were observed to evaluate the regulating effect on mRNA and protein expression of CYP.Results:After compatibility,the contents of TBA,ALP,ALT and AST decreased compared with FZ group.Meanwhile,the pathological damage induced by FZ in liver reduced in the compatibility group.The expressions of CYP1A2 and CYP3Al mRNA were inhibited in FZ group,and could be up-regulated after the combination with RS and GC.The protein levels are consistent with the the mRNA level.Conclusions:After the compatibility of RS and GC,the expression of CYP450 was up-regulated,and the accumulation time of aconite alkaloids was reduced,thus toxicity was decreased.The order of CYP450 upregulation is RS+GC+FZ>GC+FZ>RS+FZ.