人参附子及其有效成分配伍对心脏表氧化酶CYP2J3、羟化酶CYP4A3和CYP4F11 mRNA表达的影响

Effects of ginseng aconite and its effective components combination on the mRNA’s expression of oxidase CYP2J3 hydroxylase,CYP4A3 and CYP4F11

目的:考察人参及其主要成分人参皂苷Rb1,Rb2,Re,Rg1和附子及其主要成分乌头碱配伍后对CYP2J3,CYP4A3和CYP4F11 mRNA表达的影响。方法:连续灌胃(ig)给予人参水提液、附子水提液以及人参附子配伍水提液8 d后取出心脏,用real-time PCR检测各药物处理组心脏CYP2J3,CYP4A3和CYP4F11mRNA表达的改变。用不同浓度的人参皂苷Rb1,Rb2,Re,Rg1、乌头碱及其配伍药物处理H9c2细胞12,24和48 h,MTS法检测细胞的存活率,利用吸光度测定细胞的LDH来评价药物对细胞的损伤程度。人参皂苷Rb1、Re、乌头碱及其配伍药物处理H9c2细胞24和48 h,用real-time PCR检测人参皂苷Rb1、Re、乌头碱及其配伍药物对心肌细胞CYP2J3,CYP4A3和CYP4F11 mRNA表达的影响。结果:在动物实验中与空白组相比附子抑制CYP2J3 mRNA表达(P <0. 001),诱导CYP4A3和CYP4F11表达(P <0. 05);人参诱导CYP2J3mRNA的表达(P <0. 01),抑制CYP4A3和CYP4F11的作用(P <0. 001);人参附子配伍抑制CYP2J3 mRNA的表达,但结果弱于附子组,且也抑制CYP4A3和CYP4F11的表达(P <0. 001)。在细胞实验中与空白组相比乌头碱抑制CYP2J3 mRNA表达,诱导CYP4A3和CYP4F11表达。人参皂苷Rb1,Re和人参皂苷Re乌头碱配伍诱导CYP2J3 mRNA的表达,同时抑制CYP4A3和CYP4F11的表达。结论:人参及其主要单体成分可以通过改变CYP2J3,CYP4A3和CYP4F11转录水平来减少附子及乌头碱对心脏和心肌细胞的毒性。由此可以推测人参及其有效单体成分可以减少由附子及乌头碱引起心脏和心肌细胞损伤。

Objective:To investigate the effects of ginseng and the combination of its main components including ginsenoside Rb1,Rb2,Re,Rg1,and aconite and aconitine on the mRNA expression of CYP2J3,CYP4A3 and CYP4F11.Methods:SD rats were intragastrically administered ginseng aqueous extract,aconiteaqueous extract and ginseng aconite aqueous extract for 8 days and then the hearts were removed.Real-time PCR was used to detect the changes of CYP2J3,CYP4A3 and CYP4F11 mRNA expression in each drug treatment group.In the cell experiment,different concentrations of ginsenoside Rb1,Rb2,Re,Rg1,aconitine and their combinations were used to treat H9c2 cardiomyocytes for 12 h,24 h and 48 h.Cell viability was measured by MTS method,and LDH was determined by absorbance to evaluate the degree of cell damage by the drugs.Ginsenoside Rb1,Re,aconitine and their combinations were used to treat H9c2 cardiomyocytes for 24 and 48 h.Real-time PCR was used to detect ginsenoside Rb1,Re,aconitine and their combination for myocardial cells CYP2J3,CYP4A3 and CYP4F11 and the effect of mRNA expression.Results:In the animal experiment,compared with the blank group,the aconite group had an inhibitory effect on the expression of CYP2J3 mRNA(P<0.001),while CYP4A3 and CYP4F11 were induced(P<0.05).The ginseng group had an induction effect on the expression of CYP2J3 mRNA(P<0.01),and CYP4A3 and CYP4F11 were significantly inhibited(P<0.001).The ginseng aconite combination group inhibited the expression of CYP2J3 mRNA,but the results were weaker than the aconite group,and CYP4A3 and CYP4F11 were significantly inhibited(P<0.001).Compared with the blank group,the aconitine group inhibited the expression of CYP2J3 mRNA and induced the expression of CYP4A3 and CYP4F11.The combination of ginsenoside Rb1,Re and ginsenoside reconitine induced the expression of CYP2J3 mRNA and inhibited the expression of CYP4A3 and CYP4F11.Conclusion:Ginseng and its major monomeric components can reduce the toxicity of aconite and aconitine to heart by altering the transcriptional levels of CYP2J3,CYP4A3 and CYP4F11.It can be speculated that ginseng and its effective monomer components can reduce heart and H9c2 cell damage caused by aconite and aconitine.