异功散中人参皂苷类成分在脾虚大鼠模型肠吸收动力学变化及其陈皮的影响

Changes in Intestinal Absorption Kinetics of Ginsenosides in Yi Gong San in the Rat Model of Spleen Deficiency and its Effect of Pericarpium Citri Reticulatae

目的基于脾虚大鼠模型探讨异功散中陈皮对人参皂苷类成分肠吸收变化的影响及其影响是否与P-gp蛋白外排作用有关。方法分组设计加入P-gp蛋白激动剂利福平与抑制剂维拉帕米,将36只SD大鼠随机分为空白对照组(6只)与模型组(30只),予以皮下注射利血平建立脾虚模型后将模型组分为异功散组(Y组)、去陈皮异功散组(Y-C组)、异功散利福平组(Y+R组)、去陈皮异功散利福平组(Y-C+R组)、去陈皮异功散维拉帕米组(Y-C+V组),以人参皂苷Rb1、人参皂苷Re及人参皂苷Rg1为指标成分,进行在体单向肠灌流实验,采用HPLC-MS测定指标成分含量并计算肠吸收动力学参数有效渗透系数(P_(eff))与吸收速率常数(K_(a))。结果与Y组相比,Y-C组中人参皂苷Rb1和人参皂苷Re在多个灌流时段的P_(eff)显著降低,Y+R组中人参皂苷Rb1、人参皂苷Re及人参皂苷Rg1的P_(eff)和K_(a)在多个灌流时段降低,表明陈皮促进人参皂苷Rb1和人参皂苷Re肠吸收且该作用与P-gp蛋白活性有关,但未能影响人参皂苷Rg1的肠吸收;与Y-C组相比,Y-C+V组中P-gp蛋白抑制剂维拉帕米使人参皂苷Rb1和人参皂苷Re在多个灌流时段P_(eff)显著升高,而Y-C+R组中P-gp蛋白激动剂利福平使人参皂苷Rb1、人参皂苷Re及人参皂苷Rg1的P_(eff)和K_(a)在多个灌流时段显著降低,说明无陈皮干预下,人参皂苷Rb1、人参皂苷Re及人参皂苷Rg1也可能是P-gp蛋白的底物。结论异功散中陈皮通过发挥抑制P-gp蛋白活性的类维拉帕米作用来促进人参皂苷Rb1和人参皂苷Re肠吸收。

Objective To investigate the effect of Pericarpium Citri Reticulatae on changes of the intestinal absorption of ginsenoside components in Yi Gong San based on the rat model of spleen deficiency and whether its effect is related to the exocytosis of P-gp protein.Methods The group design included the P-gp protein agonist rifampicin and the inhibitor verapamil,and 36 SD rats were randomly divided into a blank control group(6 rats)and a model group(30 rats),and then subcutaneously injected with rifampicin to establish a model of splenic deficiency,and then the model group was divided into the Yigong San fullparty(Y)group,the lack of Pericarpium Citri Reticulatae(Y-C)group,the YiGong San combined with rifampin(Y+R)group,the lack of Pericarpium Citri Reticulatae combined with verapamil(YC+V)group and the lack of Pericarpium Citri Reticulatae combined with rifampin(Y-C+R)group,Ginsenoside Rb1,ginsenoside Re and ginsenoside Rg1 were used as the index components in an in situ unidirectional intestinal perfusion test,and HPLC-MS was used to determine the content of the index components and to calculate the kinetic parameters of intestinal absorption,the effective permeability coefficient(P_(eff))and the absorption rate constant(K_(a)).Results Compared with group Y,P_(eff)of ginsenoside Rb1 and Re were significantly lower in Y-C at multiple perfusion sessions,and P_(eff)and K_(a)of ginsenoside Rb1,ginsenoside Re,and ginsenoside Rg1 were lower in Y+R at multiple perfusion sessions,suggesting that Pericarpium Citri Reticulatae promotes the intestinal absorption of ginsenoside Rb1 and ginsenoside Re and that this effect is related to the activity of the P-gp protein,but failed to change the intestinal absorption of ginsenoside Rg1.The P-gp protein inhibitor verapamil significantly increased the P_(eff)of ginsenoside Rb1 and Re in the Y-C+V compared with the Y-C,while the P-gp protein agonist rifampicin significantly decreased the P_(eff)and K_(a)of ginsenoside Rb1,ginsenoside Re,and ginsenoside Rg1 in the Y-C+R in multiple perfusion sessions.The significant decrease in P_(eff)and K_(a)at multiple perfusion times suggests that ginsenoside Rb1,ginsenoside Re,and ginsenoside Rg1 may be substrates for P-gp proteins in the absence of Pericarpium Citri Reticulatae intervention.Conclusion Pericarpium Citri Reticulatae promotes the intestinal absorption of ginsenoside Rb1 and ginsenoside Re by exerting a verapamil-like effect that inhibits the activity of P-gp proteins in Yi Gong San.