基于“病证-效应-生物样本分析”方法的酸枣仁“安神”药性物质及归经的研究

Investigation on “An-shen” Drug Substances and Meridian Tropism of Semen Zizyphi Spinosae Based on “Syndrome-effect-analysis of Biological Samples”

目的:研究酸枣仁"安神"药性物质及指标性成分体内分布与归经的相关性。方法:灌胃甲状腺片混悬液160 mg·kg-1连续13天制备大鼠阴虚模型,结合高架十字迷宫装置和明暗箱装置评价酸枣仁对阴虚大鼠焦虑行为的影响;将阴虚焦虑大鼠于灌胃酸枣仁水煎液后10、20、30、40、60、90、120、240 min时摘眼球取血,并立即处死,采集心、肝、脾、肺、肾、胃、脑、大肠、小肠等组织,联合应用HPLC-PDA-ELSD检测斯皮诺素、酸枣仁皂苷A和酸枣仁皂苷B在大鼠各组织中的浓度,检测数据用DAS 2.0软件处理得到相关药动学参数。结果:与阴虚模型组比较,酸枣仁显著降低阴虚大鼠24 h内异常增高的进食量(P<0.01)和饮水量(P<0.01),显著增加实验前后体质量差值(P<0.01),显著降低肾脏系数(P<0.01)和肾上腺系数(P<0.01),显著降低T3(P<0.01)、T4(P<0.05)测定值,显著升高TSH测定值(P<0.01),表明酸枣仁可以明显改善阴虚症状;显著增加开臂进入次数百分比(P<0.01)、开臂滞留时间百分比(P<0.01)和明暗箱穿箱次数(P<0.01),表现出明显的抗焦虑作用。指标成分体内分布结果显示,斯皮诺素和酸枣仁皂苷A经血液循环广泛地分布于心、肝、脾、肺、肾、胃、脑、大肠、小肠等组织中,酸枣仁皂苷B经血液循环分布于血、胃、大肠和小肠中,其中,斯皮诺素在组织中的平均分布浓度依次为小肠>胃>肝>脑>大肠>脾>肺>心>肾,在组织中的AUC0-t依次为小肠>胃>肝>大肠>脾>脑>心>肾>肺;酸枣仁皂苷A在组织中的平均分布浓度依次为肺>大肠>心>脾>肝>肾>小肠>胃>脑,在组织中的AUC0-t依次为肺>脾>肝>心>大肠>脑>胃>肾>小肠;酸枣仁皂苷B在组织中的平均分布浓度依次为大肠>小肠>胃,在组织中的AUC0-t依次为大肠>小肠>胃。结论:酸枣仁能有效改善阴虚症状,能很好地对抗阴虚大鼠的焦虑行为,具有良好的抗焦虑作用。酸枣仁中斯皮诺素和酸枣仁皂苷A为"安神"功效物质,也是酸枣仁甘、酸味的物质基础。斯皮诺素和酸枣仁皂苷A成分在阴虚焦虑大鼠体内的分布情况与酸枣仁的归经情况比较接近。

This article was aimed to study ＂An-shen＂ drug substances of semen ziziphi spinosae and the relationship between index component distribution in vivo and meridian tropism. Intragastric administration of thyroid tablet suspension at the dose of 160 mg＇kg-1 was given for 13 days for the establishment of yin deficiency rat model. Elevated plus maze test （EPM） was combined with light/dark box test to evaluate the effect of semen zizipki spino^ae on anxiety behavior among yin deficiency rats. The yin deficiency anxiety model rats＇ eyeballs were picked for blood at 10, 20, 30, 40, 60, 90, 120, 240 rain after intragastric administration of semen ziziphi spinosae decoction. The rats were sacrificed for the collecting of heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine and small intestine and other tissues. HPLC-PDA-ELSD was combined to detect concentrations of spinosin, jujuboside A and jujuboside B in different tissues of rats. Related pharmacokinetic parameters were achieved after processing detected data with DAS 2.0 software. The results showed that compared with the yin deficiency group, semen ziziphi spinosae significantly reduced the rats＇ abnormal increased food-intake （P 〈 0.01） and water intake （P 〈 0.01） within 24 hours; significantly increased the body weight difference before and after treatment （P 〈 0.01）; significantly reduced the kidney coefficient （P 〈 0.01） and adrenal gland coefficient （P 〈 0.01）; significantly reduced the value of T3 （P 〈 0.01） and T4 （P 〈 0.05）; significantly increased the value of TSH （P 〈 0.01）. It showed that semen ziziphi spinosae can obviously improve yin deficiency symptoms. It significantly increased the number of open arms entering percentage （P 〈 0.01）, the open arms holding percentage （P 〈 0.01）, and the box through times in light/dark box test （P 〈 0.01）. It showed obvious anti-anxiety effects. The index component distribution in vivo results showed that spinosin and jujuboside A were widely distributed in the heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine, small intestine and other tissues through blood circulation. Jujuboside B was distributed in the blood, stomach, large intestine and small intestine through blood circulation. The order of average concentration of spinosin among tissues was small intestine 〉 stomach 〉 liver 〉 brain 〉 large intestine 〉 spleen 〉 lungs 〉 heart 〉 kidneys. The order of AUC0-t in tissues was small intestine 〉 stomach 〉 liver 〉 large intestine 〉 spleen 〉 brain 〉 heart 〉 kidneys 〉 lungs. The order of average concentration of jujuboside A among tissues was lungs 〉 large intestine 〉 heart 〉 spleen 〉 liver 〉 kidneys 〉 small intestine 〉 stomach 〉 brain. The order of AUC0-t in tissues was lungs 〉 spleen 〉 liver 〉 heart 〉 large intestine 〉 brain 〉 stomach 〉 kidneys 〉 small intestine. The order of average concentration of jujuhoside B among tissues was large intestine 〉 small intestine 〉 stomach. The order of AUC0-t in tissues was large intestine 〉 small intestine 〉 stomach. It was concluded that semen ziziphi spinosae can obviously improve yin deficiency symptoms with good anti-anxiety effects. Spinosin and jujuboside A in semen ziziphi spinosae were the drug substances of ＂An-shen＂ effect. They were also the material basis of sweet and sour taste. The spinosin and jujuboside A distribution in vivo of yin deficiency anxiety model rats were close to the meridian tropism of semen ziziphi spinosae.