甲氨蝶呤与长春新碱双载红细胞的制备及其生物学特性

Preparation and biological characteristics of MTX and VCR co-encapsulated erythrocytes

目的制备甲氨蝶呤(MTX)与长春新碱(VCR)双载红细胞,探讨红细胞作为双药载体的可行性。方法改良低渗预膨胀法制备MTX+VCR双载红细胞,高效液相色谱法(HPLC)分析MTX、VCR含量,自动细胞分析仪测定红细胞生理指标,考察双载红细胞的载药参数、释放情况。以渗透脆性检测、新鲜血天然免疫功能快速测定等方法,分析其贮存稳定、渗透脆性和载体红细胞天然免疫黏附功能的变化。结果联合载药情况下,不同浓度的MTX包载效果差异显著(P<0·05)。任意增加一药的浓度可增加此药的包载量而不影响另一药的包载量。1hMTX、VCR释放率分别为13·86%±0·48%和8·16%±0·26%,药物释放缓慢。双载红细胞形态和天然免疫黏附功能无明显改变,渗透脆性减低,体积略有增大,可稳定贮存5天。结论制备MTX+VCR双载红细胞基本可行,红细胞能够作为双药甚至多药的联合载体。

Objective To explore the method and feasibility of preparing MTX＋VCR-Ioaded erythrocytes using healthy adult erythro- cytes. Methods We used the modified hypotonic pre-swelling and isotonic resealing technique to prepare MTX＋VCR-loaded erythrocytes. The loading parameters and releasing rate were evaluated with high penformance liquid chromatograph （HPLC） and auto-cytoanalysis. We observed the osmotic fragility of MTX＋VCR-loaded erythrocytes conventionally. The change in innate immune activity of co-encapsulated erythrocytes was measured by counting the rosette rate of MTX＋VCR-loaded erythrocytes adhering to S180. Rmalts The concentration of MTX showed significant influence on the co-encapsulation efficiency （P〈0.05）. When concentration of MTX was fixed, the loading amount of VCR was increased according to the concentration of VCR, and vice versa. The releasing rate of MTX and VCR from MTXA＋VCR-loaded erythrocytes were （13.86±0.48）% and （8.16±0.26）% respectively at lh, and they increased gradually. The morphology and osmotic fragility of MTXA-VCR-loaded erythrocytes showed no significantly change compared with un-loaded erythrocytes. MTXA＋VCR-loaded erythrocytes could be stored stably in PBS for five days. The rosette rate of MTXA-VCR-loaded erythrocytes adhering to S180 was not significantly increased compared with un-loaded erythrocytes （P〉0.05）. Cendusiem The preparation and biological characteristics of MTX＋VCR-loaded erythrocytes were not different from those of mono-loaded erythrocytes, and erythrocytes were practically capable to be used as carriers for co-encapsulation of MIX and VCR.