五味子甲素在小鼠体内组织的分布特点及其靶向疗效研究

Distribution characteristics of Schisandrin A in mice tissues and its targeting therapeutic effect

【目的】建立小鼠组织中五味子甲素的质量浓度测定方法,并研究五味子甲素在小鼠体内的组织分布及其靶向治疗效果。【方法】将五味子甲素按25mg/kg一次灌胃ICR雄性小鼠,于给药后0.5,1,2,3和4h分别取小鼠心、肝、肾、脑,用生理盐水洗净后制成0.25g/mL组织匀浆,以睾丸酮为内标物,采用高效液相色谱法(HPLC)测定五味子甲素在小鼠心、肝、肾、脑中的质量浓度,并对HPLC方法的专属性、回收率、精密度和稳定性进行考察。依据五味子甲素在各组织中的峰值及各时间段的质量浓度,确定其分布最高的组织,对质量浓度分布最高的组织进行相关的药理学评价。【结果】小鼠心、肝、肾、脑组织中五味子甲素质量浓度为0.625~20μg/mL时,其质量浓度与五味子甲素/内标峰面积线性关系良好(r≥0.999 0)。HPLC法的专属性良好;日内精密度为(3.15±0.82)%^(4.77±0.57)%,日间精密度为(5.51±1.82)%^(8.46±2.93)%;稳定性试验的RSD<10%;绝对回收率为(89.04±3.82)%^(91.37±3.77)%,相对回收率为(95.65±5.76)%^(106.07±8.29)%。五味子甲素1次灌胃给药(剂量25mg/kg)后在小鼠体内心、肝、肾、脑组织中均有分布,给药1h后五味子甲素在小鼠心脏、脑组织中质量浓度达到峰值,给药2h后在小鼠肝脏和肾脏中质量浓度达到峰值。整个分布情况显示,给药后1,2,3h五味子甲素在小鼠肝脏组织中的质量浓度均高于其他组织,且达峰值时其质量浓度也显著高于心脏、肾脏、脑(P<0.05);其次是肾脏,五味子甲素质量浓度较低的为心脏和脑。五味子甲素对CCl4所致的小鼠急性肝损伤确有一定的保护作用。【结论】该测定方法简便、快速、稳定,可用于小鼠内脏组织中五味子甲素质量浓度的测定;小鼠灌胃给药后五味子甲素在小鼠体内心、肝、肾、脑组织中均有分布,且达峰值时以肝脏中质量浓度最高,其次是肾脏,心脏与脑中较低;五味子甲素对CCl4所致的小鼠急性肝损伤有一定保护作用。

[Objective] This study established a method to detect the concentration of Schisandrin A in mice,investigated its distribution characteristics and examined its targeting therapeutic effect. [Method] Schisandrin A at concentration of 25 mg/kg was administered orally to ICR mice and the concentrations in heart, brain,kidney and liver were measured using HPLC with testosterone as internal standard 0.5,1,2, 3,and 4 h after administration. The specificity,recovery,accuracy and stability of HPLC method were also studied. The tissue with highest Schisandrin A contribution was determined and the biological activities were evaluated. [Result] The concentrations of Schisandrin A distribution in mice heart, liver, kidney, and brain tissues were 0. 625 to 20 μg/mL,having a good linear relationship with the internal standard peak area ratio （r≥0. 999 0）. HPLC had a good specificity,with within-day precision of （3. 15 ±0. 82）%- （4.77±0.57）% and between-day precision of （5.51±1.82）%- （8.46±2.93）%. The RSD of stability test was 〈10% ,the absolute recovery was （89.04±3.82）%- （91.37±3.77）% ,and the relative recovery was （95.65±5.76）%- （106. 07±8.29）%. Sehisandrin A was distributed in heart,liver,kidney and brain, and the times needed to reach peak concentration were 1 hour in heart and brain,and 2 hours in liver,and 3 hours in kidney. The concentration of Schisandrin A in liver was significantly higher than in kidney,heart and brain （P〈0.05）. Schisandrin A had protective effect on CC14 induced acute liver injury in mice. [Conclusion] The established method was simple, rapid, stable, and can be used for determination of Schisandrin A in internal tissues of mice. After oral administration,peak and every time concentrations of Schisandrin A were highest in liver,and it can protect acute liver injury.