姜炭在虚寒性出血证大鼠体内的整合药代动力学与温经止血药效学的相关性研究

Correlation between integrated pharmacokinetics and pharmacodynamics of warming channel for arresting bleeding in ginger charcoal treated hemorrhagic rats with deficiency-cold syndrome

目的:探讨姜炭在虚寒性出血证大鼠体内的整合药代动力学与温经止血药效学之间的相关性。方法:18只雄性SD大鼠随机分为空白组、模型组和姜炭组,每组6只。模型组和姜炭组大鼠诱导虚寒性出血证模型。造模同时,姜炭组大鼠灌胃给予0.007 g/kg姜炭乙酸乙酯提取液,空白组和模型组大鼠灌胃给予等体积蒸馏水,连续5 d。末次给药后分别于0.083、0.25、0.5、0.75、1、2、3、4、6、8、10、12 h采集血样。高效液相色谱(HPLC)法测定各组大鼠不同时间点血清中6-姜烯酮、6-姜烯酚、姜酮、二乙酰基-6-姜酚、6-姜酚、10-姜酚的含量。基于自定义权重系数法建立整合药代动力学模型,并计算整合药动学参数。酶联免疫吸附法(ELISA)检测血清血栓素B2(TXB2)和6-酮-前列腺素F1α(6-keto-PGFlα)的含量,并计算药效强度。对整合血药浓度与药效强度进行相关性分析。结果:①建立整合药代动力学模型,计算药动学参数半衰期(t1/2)为(3.59±0.20)h,达峰时间(Tmax)为(3.00±0.00)h,最大浓度(Cmax)为(6.34±0.24)μg/ml,曲线下面积(AUC0-∞)为(44.44±1.41)μg·h/ml,平均滞留时间(MRT)为(5.85±0.18)h,表观清除率(CL/F)为(47.29±1.49)ml/(mg·h)。②与空白组相比,模型组大鼠各时间点血清TXB2含量及TXB2/6-keto-PGFlα比值均显著降低(P<0.05)。与模型组相比,姜炭组大鼠各时间点血清TXB2含量及TXB2/6-keto-PGFlα比值均显著升高(P<0.05),且药效强度较高。③多成分整合血药浓度与血清TXB2和TXB2/6-keto-PGFlα的药效强度变化趋势基本一致,给药4 h后整合血药浓度与药效强度呈正相关。结论:姜炭药效组分在虚寒性出血证大鼠体内的整合药代动力学与温经止血的药效学具有一定的相关性。

Objective: To investigate the correlation between integrated pharmacokinetics and pharmacodynamics of warming channel for arresting bleeding in ginger charcoal treated hemorrhagic rats with deficiency-cold syndrome. Methods: Eighteen male SD rats were randomly divided into the blank group,model group and ginger charcoal group,6 rats in each group. The hemorrhage model with deficiency-cold syndrome was induced in the model group and ginger charcoal group. Meanwhile,the rats in the ginger charcoal group were treated with ethyl acetate extract solution of ginger charcoal by intragastric administration at dose of 0.007 g/kg,the rats in the blank group and model group were treated with distilled water at equal volume,with a course of 5 days. After the last administration,the blood samples were collected at 0.083,0.25,0.5,0.75,1,2,3,4,6,8,10 and 12 h,respectively. The serum contents of 6-gingerone,6-shogaol,gingerone,diacetyl-6-gingerol,6-gingerol and 10-gingerol in rats of each group were determined at different time points by high performance liquid chromatography(HPLC). The integrated pharmacokinetic model was established based on custom weight coefficient,and the integrated pharmacokinetic parameters were calculated. The serum contents of thromboxane B2(TXB2) and 6-ketoprostaglandin F1α(6-keto-PGFlα) were detected by enzyme-linked immunosorbent assay(ELISA),and the pharmacodynamic intensity was calculated. The correlation between the integrated concentrations and the pharmacodynamic intensity was analyzed. Results:(1)The integrated pharmacokinetic model was established. The half-life(t1/2) was(3.59±0.20) h,the peak time(Tm ax) was(3.00±0.00) h,the maximum concentration(Cm ax) was(6.34±0.24) μg/ml,the area under the curve(AUC0-∞) was(44.44±1.41) μg·h/ml,the mean retention time(MRT) was(5. 85 ± 0. 18) h,and the apparent clearance rate(CL/F) was(47. 29 ± 1. 49) ml/(mg ·h).(2)Compared with the blank group,the serum content of TXB2 and the ratio of TXB2 to 6-keto-PGFlαwere significantly reduced in the model group at all time points(P<0.05). Compared with the model group,the serum content of TXB2 and the ratio of TXB2 to 6-ketoPGFlαwere significantly increased in the ginger charcoal group at all time points(P<0.05),with higher pharmacodynamic intensity.(3)The change of integrated concentration of multi-components was consistent with the change of pharmacodynamic intensity in serum TXB2 and TXB2/6-keto-PGF lαratio,and there was a positive correlation between the integrated blood concentration and the pharmacodynamic intensity after administration for 4 hours. Conclusion: There is a certain correlation between integrated pharmacokinetics and pharmacodynamics of warming channel for arresting bleeding in ginger charcoal treated hemorrhagic rats with deficiency-cold syndrome.