| 荧光分析法测定聚乙二醇化脂质体的包封率及其药动学研究 |
| 投稿时间：2025-02-12 修订日期：2025-02-26 点此下载全文 |
| 引用本文：潘建全,张雪,张成,邓慧雅,王凤玲,陈卫东.荧光分析法测定聚乙二醇化脂质体的包封率及其药动学研究[J].医学研究杂志,2025,54(7):47-53, 5 |
| DOI：
10.11969/j.issn.1673-548X.2025.07.010 |
| 摘要点击次数: 4 |
| 全文下载次数: 5 |
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| 基金项目:国家自然科学基金资助项目(面上项目)(82073923) |
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| 中文摘要:目的 探究荧光分析法测定聚乙二醇化脂质体(PEGylated liposome,PEG-L)的包封率及其用于加速血液清除(accelerated blood clearance,ABC)现象研究的可行性。方法 采用乙醇注入法制备出荧光染料DiR标记的PEG-L(PEGylated liposomal DiR,PEG-DiR-L),对PEG-DiR-L分别通过超滤离心法和超声破乳法获得游离DiR样品和总DiR样品；使用多功能酶标仪测定两种样品中DiR的含量,进而计算出PEG-DiR-L的包封率。将雄性Sprague-Dawley(SD)大鼠分为单次注射组和重复注射组,单次注射组首次注射磷酸盐缓冲液(phosphate buffer solution,PBS),间隔7天后再次注射PEG-DiR-L,重复给药组首次注射PEG-L,间隔7天后再次注射PEG-DiR-L。对大鼠进行眼眦取血,利用多功能酶标仪测定PEG-DiR-L的相对血药浓度,绘制药动学曲线,计算主要药动学参数,比较两组大鼠血药浓度的变化。采集大鼠的心脏、肝脏、脾脏、肺和肾脏组织,采用小动物活体成像仪进行离体成像,比较PEG-DiR-L在两组大鼠不同组织中的生物分布情况。结果超滤离心法测得PEG-DiR-L的包封率均值为97.68%±0.23%；与单次注射组比较,重复注射组中的血药浓度降低速率增加,AUC(0→t)、AUC(0→∞)、MRT(0→t)、MRT(0→∞)和t1/2z降低,CLz和Vz增加(P < 0.05),PEG-DiR-L的肝组织蓄积增加。结论血浆药动学和组织分布结果表明,重复注射组发生了ABC现象,说明荧光分析法能够测定PEG-L的包封率并用于研究ABC现象。 |
| 中文关键词:加速血液清除现象 聚乙二醇化脂质体 荧光分析法 |
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| Fluorescence Analysis Method for Determining the Encapsulation Rate of PEGylated Liposomes and Their Pharmacokinetics. |
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| Abstract:Objective To investigate the feasibility of fluorescence analysis method for determining the encapsulation efficiency (EE) of PEGylated liposomes (PEG-L) and their application in studying the accelerated blood clearance (ABC) phenomenon. Methods DiR-labeled PEGylated liposomes (PEG-DiR-L) were prepared via ethanol injection. Free DiR and total DiR samples were obtained using ultrafiltration centrifugation and ultrasonic demulsification, respectively. The EE of PEG-DiR-L was calculated by quantifying DiR levels in both samples using a multifunctional microplate reader. Male Sprague-Dawley (SD) rats were divided into two groups:the single injection group [first injected with phosphate-buffered saline (PBS), followed by PEG-DiR-L 7days later] and the repeated injection group (first injected with empty PEG-L, followed by PEG-DiR-L 7days later). Blood samples were collected from the eye canthus of rats, and the plasma concentrations of PEG-DiR-L were measured to construct pharmacokinetic profiles and calculate key pharmacokinetic parameters. Tissue distribution was assessed by ex vivo imaging of heart, liver, spleen, lung, and kidney samples using the IVIS spectroscopy. Results The EE of PEG-DiR-L determined by ultrafiltration centrifugation was 97.68%±0.23%. Compared to the single-dose group, the repeated-dose group exhibited significantly accelerated plasma clearance, with reduced AUC(0→t)、AUC(0→∞)、MRT(0→t)、MRT(0→∞) and t1/2z, alongside increased CLz and Vz (P < 0.05). Hepatic accumulation of PEG-DiR-L was markedly enhanced in the repeated-dose group. Conclusion The results of plasma pharmacokinetics and tissue distribution indicated that the ABC phenomenon occurred in the repeated injection group, suggesting that the fluorescence analysis method is able to determine the encapsulation rate of PEG-L and be used to study the ABC phenomenon. |
| keywords:Accelerated blood clearance phenomenon PEGylated liposomes Fluorescence method |
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