目的 研究谷红注射液中羟基红花黄色素A(HSYA)在脑缺血再灌注大鼠体内的药动学及其与抗氧化作用的关联性。方法 采用平衡透析法测定HSYA及谷红注射液中HSYA的血浆蛋白结合率;SD大鼠制备大脑右侧中动脉局灶性栓塞(MCAO)模型,分为HSYA组(4 mg/kg)及谷红注射液组(10 mL/kg),尾iv给药,采用高效液相(HPLC)法测定不同时间点HSYA的血药浓度,绘制药时曲线;同时采用试剂盒法测定不同时间点血浆中谷胱甘肽过氧化物酶(GSH-Px)、乳酸脱氢酶(LDH)活性,绘制效时曲线;进一步进行药动-药效联动分析。结果 HSYA和谷红注射液中HSYA在质量浓度为2.5、10.0、25.0 mg/L时血浆蛋白结合率分别是77.96%、73.54%、76.13%和68.21%、58.22%、63.17%;HSYA的血浆药物质量浓度在0.01～50 mg/L线性关系良好,低、中、高血浆药物浓度平均回收率分别为(99.94±2.82)%、(104.16±1.41)%、(99.74±1.06)%;谷红注射液组与HSYA组比较,曲线下面积(AUC)显著增加,平均驻留时间(MRT_0-t)显著减少,表观容积(V_z)显著增加;血浆中GSH-Px活性升高,与血药浓度呈正相关,LDH活性降低,与血药浓度呈负相关。结论 HSYA均具有中等强度的血浆蛋白结合率,且谷红注射液中HSYA血浆蛋白结合率有所降低;谷红注射液在MCAO大鼠中可以增加HSYA的生物利用度,提高药物的效果,增加药物在体内的分布;谷红注射液组与HSYA组比较具有更好的抗氧化作用,对大鼠脑缺血再灌注损伤有明显的保护作用。

Objective To investigate the in vivo pharmacokinetic progress of hydroxysafflor yellow A (HSYA) from Guhong Injection in cerebal ischemia reperfusion (I/R) injury of rats and the correlation with its anti-oxidation effect. Methods The equilibrium dialysis method was carried out to determine the plasma protein binding rates of HYSA and HSYA in Guhong Injection. Middle cerebral artery occlusion (MCAO) model rats were iv injected HYSA (4 mg/kg) or Guhong Injection (10 mL/kg). The HPLC method was adopted to determine the plasma concentration of HYSA at different time points to draw the drug-time curve. Meanwhile, glutathione peroxidase (GSH-Px) and lactate dehydrogenase (LDH) activities were determined to draw the time-effect curve. Furthermore, the relationship between pharmacokinetics and pharmacodynamics was analyzed. Results At the concentration of 2.5, 10, and 25 mg/L, the p plasma rotein binding rates of HYSA were 77.96%, 73.54%, and 76.13%, whereas the plasma protein binding rates of HYSA from Guhong Injection were 68.21%, 58.22%, and 63.17%, respectively. A good linear relationship of HYSA was obtained in the range of 0.01—50 mg/L, the mean recoveries were (99.94 ± 2.82)%, (104.16 ± 1.41)%, and (99.74 ± 1.06)% for low, middle, and high concentration of the samples, respectively. Compared with HYSA group, Guhong Injection significantly increased the AUC of HYSA and decreased the MRT and V_z of HYSA. Furthermore, Guhong Injection increased the content of GSH-Px and decreased the content of LDH. The plasma concentration of HYSA is positively related to the GSH-Px activity and negatively related to the LDH activity. Conclusion The results indicate that HYSA has the moderate plasma protein binding rate. Compared with HYSA group, the plasma protein binding rate in Guhong Injection group is reduced. Guhong Injection could increase the bioavailability of HYSA to enhance therapeutic efficacy and increase the distribution of HYSA in ischemia rats. Guhong Injection has better anti-oxidant effect, as well as more significant protective effect against cerebral I/R injury than HYSA.

国家自然科学基金资助项目(81373898,81173647);浙江省卫生高层次创新人才培养工程项目