心肌缺血再灌注损伤大鼠体内荭草花活性成分的PK-PD研究

Study on PK-PD of active components of Polygonum orientale L.flower in rats with myocardial ischemia-reperfusion injury

目的建立药代动力学-药效动力学(PK-PD)结合模型,探讨荭草花提取物在心肌缺血再灌注损伤(MIRI)模型大鼠体内活性成分的动态变化与其药效消长之间的关系。方法荭草花药材经水煮醇沉、正丁醇萃取得荭草花提取物;取SD大鼠6只,采用冠脉结扎法制备MIRI模型,术前大鼠灌胃荭草花提取物86 g/kg,3次/d、连续5 d,于末次给药后5 min、10 min、15 min、30 min、1.0 h、1.5 h、2.0 h、4.0 h、6.0 h、8.0 h、10.0 h、12.0 h、24.0 h、36.0 h及48.0 h经尾静脉取血,采用高效液相色谱-串联质谱(HPLC-MS/MS)检测血浆样本中6种活性成分(原儿茶酸、槲皮苷、花旗松素、N-p香豆酰酪胺、山奈素-3-O-β-D-葡萄糖苷及山奈素-3-O-α-L-鼠李糖苷)的质量浓度,获取各成分血药浓度-时间曲线;采用试剂盒测定血浆样本中超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)及心肌钙蛋白I(cTn-I)的浓度,获取效应-时间曲线;通过Winnonlin 6.4软件分别对药物浓度-时间与效应-时间进行拟合,建立PK-PD模型。结果荭草花提取物中各有效成分均能够较好地与各药效指标拟合,以SOD为药效指标时,各成分的PK-PD模型以Sigmoid E_(max)拟合较优;以LDH、CK-MB及cTn-I为药效指标时,各成分的PK-PD模型以Inhibitory Effect拟合较优。结论荭草花提取物中的原儿茶酸、槲皮苷、山奈素-3-O-β-D-葡萄糖苷及山奈素-3-O-α-L-鼠李糖苷、花旗松素、N-p香豆酰酪胺具有心肌缺血保护作用,其浓度与药效指标SOD、LDH、CK-MB及cTn-I的水平有相关性。

Objective To establish a pharmacokinetic-pharmacodynamic(PK-PD)combined model and explore the correlation between dynamic changes of the active components of Polygonum orientale L.flower extract in rats with myocardial ischemia-reperfusion injury(MIRI)model and the decline of its efficacy.Methods Polygonum orientale L.Flower extract was extracted by n-butanol after water extracting and alcohol precipitation.Six SD rats were used to establish MIRI model by coronary artery ligation.Before operation,the rats were given Polygonum orientale flower extract(86 g/kg),3 times a day for continuous5 days.Blood samples were collected from tail vein at 5 min,10 min,15 min,30 min,1.0 h,1.5 h,2.0 h,4.0 h,6.0 h,8.0 h,10.0 h,12.0 h,24.0 h,36.0 h,48.0 h after the last administration.High performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was used to measure the mass concentrations of 6 active components(protocatechuate,quercitrin,taxifolin,paprazine,kaempferol-3-O-β-D-glucopyranoside and kaempferol 3-O-α-L-rhamnoside)in plasma samples.The blood drug concentration-time curves of each component were obtained.Kits were used to measure the concentrations of superoxide dismutase(SOD),lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),and cardiac troponin I(cTn-I)in plasma samples to plot the effect-time curves.The drug concentration-time and effect-time were fitted by Winnonlin 6.4 software to establish PK-PD model.Results The active components in the Polygonum orientale L.Flower were well-fitted with each pharmacodynamic index.When SOD was used as a pharmacodynamic index,PK-PD binding model was well-fitted with Sigmoid E_(max).When the pharmacodynamic indexes were LDH,CK-MB,and cTn-I,PK-PD binding model was well-fitted by inhibitory effect.Conclusion Protocatechuate,quercitrin,taxifolin,paprazine,kaempferol-3-O-β-D-glucopyranoside,and kaempferol 3-O-α-L-rhamnoside in Polygonum orientale L.Flower extract exert protective effects on MIRI.The concentrations of 6 active components are correlated with the levels of SOD,LDH,CK-MB,and cTn-I.