摘要
目的:探讨一种氯膦酸二钠脂质体灌肠剂的制备及其诱导巨噬细胞凋亡的作用。方法:以DPPC和胆固醇为主要组分并添加带负电荷的磷酸二鲸蜡酯,用薄膜水化法制备脂质体;用MTT法测定该氯膦酸二钠脂质体对离体培养的小鼠巨噬细胞的致死活性;以免疫印迹法和流式细胞术对该氯膦酸二钠脂质体诱导的细胞凋亡进行了研究。结果:薄膜水化法制备所得的脂质体粒径适于巨噬细胞吞噬。用离体培养的小鼠巨噬细胞对氯膦酸二钠脂质体灌肠剂的细胞致死活性进行评估,该氯膦酸二钠脂质体制剂可有效地杀伤巨噬细胞,其细胞毒性随浓度而增加;免疫印迹法可从氯膦酸二钠脂质体处理的细胞提取物中检测到显著增加的活性半胱天冬酶-3,表明氯膦酸二钠脂质体可诱导细胞凋亡;流式细胞术检测Annexin V的结果确证氯膦酸二钠脂质体可诱导巨噬细胞凋亡。结论:该氯膦酸二钠脂质体制剂可诱导巨噬细胞凋亡从而清除巨噬细胞,有望用于某些免疫性疾病和炎症性疾病的辅助治疗。
Aim: To investigate apoptotic effect of a novel clodronate liposomal enema. Methods: A novel clodronate liposomal enema was developed by thin-film hydration using DPPC, cholesterol, and negatively charged dicetylphosphate. Lethality of clodronate liposomes was determined by MTT assay using RAW264.7 murine macrophages. Clodronate liposome-induced apoptosis was investigated by Western blotting and fluorescence-activated cell sorting (FACS). Results: A novel clodronate liposomal enema was developed by thin-film hydration. This liposomal enema had a size range that is suitable for phagocytosis. MTT assay showed that this clodronate liposomal enema efficiently caused cytotoxicity of RAW264.7 murine macrophages in a dose-dependent manner. Western blots showed remarkably increased cleaved caspase-3 in clodronate liposome-treated RAW264.7 cells compared to controls, indicating apoptosis induced by clodronate liposomes. In addition, FACS analysis for Annexin V confirmed that clodronate liposomes induced apoptosis in RAW264.7 macrophages. Conclusion: This clodronate liposomal enema is capable of inducing macrophage apoptosis, and may be used for macrophage depletion in certain diseases as an adjuvant therapy.
出处
《世界肿瘤研究》
2017年第4期90-98,共9页
World Journal of Cancer Research
基金
兰州市人才创新创业项目(2014-RC-81)
人社部2015年留学人员回国创业启动支持计划资助。
