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目的研究L02细胞对白蛋白纳米粒的胞吞作用。方法采用钙黄绿素作为白蛋白纳米粒的荧光标记探针,用去溶剂化法制备钙黄绿素白蛋白纳米粒(calcein HSA-NPs),并考察粒径、体外释放、细胞摄取、胞内分布和细胞毒性。结果 CalceinHSA-NPs平均粒径为203.3 nm,体外释放96 h不到80%。白蛋白纳米粒的胞吞作用呈现时间、浓度、温度依赖性,可以显著提高水溶性小分子钙黄绿素的入胞能力(P<0.01)。Calcein HSA-NPs主要经clathrin介导的内吞途径和大胞饮入胞的,进入细胞后分布在细胞核外的胞内空间,主要集中在溶酶体内。白蛋白纳米载体对细胞基本无毒。结论所制备的calcein HSA-NPs可作为有效的载体来研究胞吞作用机制。
Objective To study the endocytosis of albumin nanoparticles by L02 cells. Methods Calcein HSA-NPs were prepared by desolvation using calcein as a fluorescent probe for albumin nanoparticles. The effects of calcein HSA-NPs on the particle size, in vitro release, cellular uptake, intracellular distribution, toxicity. Results The average diameter of Calcein HSA-NPs was 203.3 nm, and the release rate was less than 80% in 96 h. Albumin nanoparticles showed time, concentration and temperature dependency of endocytosis, which could significantly increase the ability of water-soluble small molecule calcein (P <0.01). Calcein HSA-NPs mainly by clathrin-mediated endocytosis pathway and macrophage into the cell, enter the cell distribution in the extracellular nucleus of the intracellular space, mainly concentrated in the lysosome. Albumin nanocarriers are essentially non-toxic to cells. Conclusion The prepared calcein HSA-NPs can be used as an effective carrier to study the mechanism of endocytosis.