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芹菜素纳米混悬剂在小鼠体内的组织分布研究 Δ

徐世一 ，吕 畅 ，霍元子 ，郝若祎 ，阎雪莹 （1.黑龙江中医药大学实验实训中心，哈尔滨 150040；2.齐齐哈
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尔医学院附属第三医院药剂科，黑龙江 齐齐哈尔 161099；3.黑龙江中医药大学药学院，哈尔滨 150040）
中图分类号 R969.1 文献标志码 A 文章编号 1001-0408（2020）04-0457-07
DOI 10.6039/j.issn.1001-0408.2020.04.13
摘 要 目的：研究芹菜素纳米混悬剂（AP-NPs）在小鼠体内的分布特点及靶向特性。方法：采用超声微量沉淀法制备AP-NPs。
将昆明种小鼠随机分为芹菜素（AP）溶液组和AP-NPs混悬液组，各45只。各组小鼠单次灌胃相应药物溶液（80 mg/kg），分别于给
药前（0 h）以及给药后0.25、0.5、1、2、4、6、8、10 h摘取眼球取血500 μL，并于末次取血后处死，取其心、肝、脾、肺、肾、脑组织。血浆
和各组织样品分别经甲醇沉淀蛋白后，采用高效液相色谱法测定。色谱柱为Shimadzu ODS-SP，流动相为甲醇-水（70 ∶ 30，V/V），
检测波长为340 nm，柱温为35 ℃，流速为1.0 mL/min，进样量为20 μL。根据标准曲线法计算AP在不同样品中的质量浓度，采用
DAS 2.0软件和Excel 2010软件计算AP的主要药动学参数（AUC、cmax等）和峰浓度比值（ce ）、相对摄取率（RUE）、摄取占比及其改
变值，以分析其组织分布及靶向特性。结果：AP 在血浆和各组织中检测质量浓度的线性范围均为 0.1～25.0 μg/mL（r 均大于
0.99），定量下限均为0.1 μg/mL；日内、日间RSD均小于15％，准确度为94.37％～117.48％，提取回收率均大于80％。与AP溶液
组比较，AP-NPs混悬液组小鼠血浆（0.5～6 h各时间点）以及肝（0.25～8 h各时间点）、脾（0.25～8 h各时间点）、脑（0.25～4 h各时
间点）组织样品中 AP 的质量浓度均显著升高（P＜0.05 或 P＜0.01），且以肝中含量最高；而心脏（6 h）、肝（10 h）、肺（0.5 h）、肾
（0.25～10 h各时间点）组织样品中AP的质量浓度则显著降低（P＜0.05或P＜0.01）。两组小鼠血浆/组织样品中AP的AUC、cmax比
较，差异均有统计学意义（P＜0.05），且肝组织的 ce、RUE、摄取占比及其改变值均最大，分别为 1.34±0.40、1.99±0.29、48.49％、
15.71％。结论：将AP制成纳米混悬剂后，药物的组织分布有所改变，尤其对肝组织的靶向作用有所提高。
关键词 芹菜素；纳米混悬剂；组织分布；靶向；小鼠；高效液相色谱法

Study on Tissue Distribution of Apigenin Nanosuspension in Mice
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XU Shiyi ，LYU Chang ，HUO Yuanzi ，HAO Ruoyi ，YAN Xueying（1. Laboratory Training Center，Heilongjiang
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University of Chinese Medicine，Harbin 150040，China；2. Dept. of Pharmacy，the Third Affiliated Hospital of
Qiqihar Medical College，Heilongjiang Qiqihar 161099，China；3. College of Pharmacy，Heilongjiang University
of Chinese Medicine，Harbin 150040，China）
ABSTRACT OBJECTIVE：To study the distribution and targeting characteristics of Apigenin nanosuspension（AP-NPs）in mice.
METHODS：AP-NPs was prepared with ultrasound microprecipitation. Kunming mice were randomly divided into apigenin（AP）
solution group and AP-NPs suspension group，with 45 mice in each group. The mice were given relevant medicine intragastrically
（80 mg/kg）；blood sample of eyeball 500 μL were collected before medication（0 h）and 0.25，0.5，1，2，4，6，8，10 h after
medication. After the last blood collection，the mice were sacrificed and their heart，liver，spleen，lung，kidney and brain tissues
were taken. After protein precipitation with methanol，HPLC method was adopted for determining plasma and tissues. The
determination was performed on Shimadzu ODS-SP column with mobile phase consisted of methanol-water（70 ∶ 30，V/V）at the
flow rate of 1.0 mL/min. The detection wavelength was set at 340 nm，and column temperature was 35 ℃. The sample size was 20
μL. The concentration of AP in different samples was calculated according to standard curve，main pharmacokinetic parameters
（AUC，cmax ）of AP and the ratio of peak concentration（ce ），relative uptake rate（RUE），uptake ratio and its change value were
calculated with DAS 2.0 software and Excel 2010 software；the tissue distribution and targeting characteristics of AP were analyzed.
RESULTS：The linear range of AP in plasma and tissues were 0.1-25.0 μg/mL（all r＞0.99）；the lower limits of quantification
were 0.1 μ g/mL. RSDs of intra-day and inter-day were all lower than 15％ ，and the accuracy were 94.37％ -117.48％ . The
extraction recovery rates were all more than 80％. Compared with AP solution group，the concentrations of AP in plasma sample
（during 0.5-6 h），liver tissue （during 0.25-8 h），spleen tissue （during 0.25-8 h） and cerebral tissue （during 0.25-4 h） were
increased significantly in AP-NPs suspension group（P＜0.05 or P＜0.01），and the highest in liver tissue. The concentrations of AP
in heart tiusse（6 h），liver tissue（10 h），lung tissue（0.5 h），spleen tissue（during 0.25-10 h）were decreased significantly（P＜
0.05 or P＜0.01）. There was statistical significance in AUC and cmax of AP in plasma and tissue samples between 2 groups（P＜
0.05）. The ce，RUE，uptake ratio and its change value of liver tissue were the highest，being 1.34±0.40，1.99±0.29，48.49％ and
15.71％ . CONCLUSIONS：After AP is made into nanosus-
Δ 基金项目：黑龙江省自然科学基金（重点项目）（No.ZD2016019）
pension，the distribution of drug tissue is changed，especially
＊助理实验师。研究方向：缓控释、靶向制剂及抗肿瘤制剂。电
targeting effect on liver tissue is improved.
话：0451-82196224。E-mail：994010246@qq.com
KEYWORDS Apigenin； Nanosuspension；Tissue distribu-
# 通信作者：教授，博士。研究方向：靶向、缓控释制剂的研究及
tion；Targeting；Mice；HPLC
药物的体内代谢。电话：0451-82196181。E-mail：15159267@qq.com
中国药房 2020年第31卷第4期 China Pharmacy 2020 Vol. 31 No. 4 ·457 ·