氯膦酸二钠脂质体不同给药剂量和方式对骨髓巨噬细胞清除效果的研究

Research of effect of different doses and modes of liposome administration on the clearance of bone marrow macrophages

摘要:

目的 研究氯膦酸二钠脂质体不同给药剂量和方式对骨髓巨噬细胞的清除效果。方法 用雄性BALB/c和C57BL/6小鼠复制异基因造血干细胞移植（HSCT）模型，随机分为HSCT组、HSCT + ip100组（HSCT+氯膦酸二钠脂质体腹腔注射100 μL）、HSCT + iv100组（HSCT+氯膦酸二钠脂质体静脉注射100 μL）、HSCT + iv150组（HSCT+氯膦酸二钠脂质体静脉注射150 μL）、HSCT + iv200组（HSCT+氯膦酸二钠脂质体静脉注射200 μL）、HSCT + iv250组（HSCT+氯膦酸二钠脂质体静脉注射250 μL）。HSCT后第1天注射氯膦酸二钠脂质体，之后每5天注射1次，直至相应时间点获取样本。采用流式细胞术和Western blotting检测骨髓巨噬细胞数量的变化，依据全血细胞计数确定外周血的变化，HE染色观察骨髓腔损伤。结果 与HSCT组比较，其余各组小鼠的外周血白细胞计数明显增加（P <0.05）。与氯膦酸二钠脂质体腹腔注射相比，静脉注射骨髓巨噬细胞清除效果明显（P <0.05）。流式细胞术结果显示，氯膦酸二钠脂质体静脉注射清除效果随着脂质体剂量的增加而更加明显。HE染色结果显示，随着氯膦酸二钠脂质体剂量的增加，骨髓腔中的脂肪空泡越来越多。结论 氯膦酸二钠脂质体静脉注射清除骨髓巨噬细胞优于腹腔注射，在一定剂量内，骨髓巨噬细胞随着氯膦酸二钠脂质体注射剂量的增加，清除效果越明显。

Abstract:

Objective To study the effects of different dosages and methods of Clodronate liposomes administration on the clearance of bone marrow macrophages.Methods Male BALB/c and females C57BL/6 mice were firstly used to establish an allogeneic hematopoietic stem cell transplantation (HSCT) model, and were randomly divided into HSCT group, HSCT+ Clodronate liposome 100 μL group (ip), HSCT + Clodronate Liposome 100 μL group (iv), HSCT + Clodronate Liposome 150 μL group (iv), HSCT + Clodronate Liposome 200 μL (iv), HSCT + Clodronate Liposome 250 μLgroup (iv). Mice were injected with different dose of Clodronate Liposome day 1 post HSCT and then every 5 days until corresponding time point. The changes in the number of macrophages in bone marrow were detected by flow cytometry and Western blot. The changes in peripheral blood were determined by complete blood count, and the injuries in bone marrow cavity were observed by H&E staining.Results Compared with the HSCT group, the peripheral blood leukocyte counts of the other groups were significantly increased (P < 0.05); Compared with the intraperitoneal injection group, the intravenous injection group showed a significant enhancement on the clearance of bone marrow macrophages (P < 0.05); Flow cytometry results showed that the clearance effect on bone marrow macrophages became stronger as the dose of Clodronate liposome increased. Bone marrow pathology staining showed that the fat vacuoles in the bone marrow lumen increased more with the increase of dose of Clodronate liposome.Conclusion Compared with intraperitoneal injection, intravenous injection of Clodronate liposome showed a more significant bone marrow macrophage clearance effect, and the clearance effect of clodronate liposome on bone marrow macrophages increasing with a higher dosage in a certain dose range.