Remote loading of doxorubicin into liposomes by transmembrane pH gradient to reduce toxicity toward H9c2 cells.
Remote loading of doxorubicin into liposomes by transmembrane pH gradient to reduce toxicity toward H9c2 cells.
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DOI：
10.1016/j.jsps.2015.02.014
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发表时间：
2016-03
期刊：
影响因子：
--
通讯作者：
Lahouel M
中科院分区：
文献类型：
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作者：
Alyane M;Barratt G;Lahouel M
阿霉素（DOX）的使用受到其剂量依赖性心脏毒性的限制。脂质体中包封的DOX已显示出降低心脏毒性。结果表明，脂质体包封率约占总药物的92%。释放实验表明，在培养基和PBS中的弱DOX泄漏，超过98%和90%的包封的DOX仍然分别保留在脂质体中孵育24小时后。在pH5.3的磷酸盐缓冲液中进行释放实验时，孵育24 h后DOX从脂质体中的泄漏达到37%。对脂质体DOX的细胞摄取的评估表明脂质体的可能的内吞作用，因为DOX的大部分可见荧光主要在细胞质中，而核隔室显示弱强度。当使用未负载的荧光脂质体时，荧光在核中不存在，表明脂质体不能穿过核膜。MTT法和LDH释放量测定结果表明，高剂量DOX作用下H9c2细胞以坏死为主，而低剂量DOX作用下细胞以凋亡为主。包封的DOX在孵育3和6 h后明显降低DOX的毒性，但在孵育20 h后，包封的DOX比游离的DOX的毒性更大。
The use of doxorubicin (DOX) is limited by its dose-dependent cardiotoxicity. Entrapped DOX in liposome has been shown to reduce cardiotoxicity. Results showed that about 92% of the total drug was encapsulated in liposome. The release experiments showed a weak DOX leakage in both culture medium and in PBS, more than 98% and 90% of the encapsulated DOX respectively was still retained in liposomes after 24 h of incubation. When the release experiments were carried out in phosphate buffer pH5.3, the leakage of DOX from liposomes reached 37% after 24 h of incubation. Evaluation of cellular uptake of the liposomal DOX indicated the possible endocytosis of liposomes because the majority of visible fluorescence of DOX was mainly in the cytoplasm, whereas the nuclear compartment showed a weak intensity. When using unloaded fluorescent-liposomes, the fluorescence was absent in nuclei suggests that liposomes cannot cross the nuclear membrane. MTT assay and measurement of LDH release suggest that necrosis is the form of cellular death predominates in H9c2 cells exposed to high doses of DOX, while for weak doses apoptosis could be the predominate form. Entrapped DOX reduced significantly DOX toxicity after 3 and 6 h of incubation, but after 20 h entrapped DOX is more toxic than free one.