光敏蛋白脂质体的制备与生物活性

doi:10.3864/j.issn.0578-1752.2013.01.008

摘要/Abstract

摘要： 【目的】获得KillerRed蛋白（KRP）脂质体的制备配方，测定其物理化学性质及生物活性，为优化该蛋白的使用性能提供依据。【方法】用逆相蒸发法制备KRP脂质体，用荧光分光光度法测定脂质体包封率，以包封率为指标通过正交试验优化配方，并考察脂质体的形态、粒径、Zeta电位和泄漏率以及生物活性。【结果】按优化配方制得的脂质体颗粒呈球形，平均粒径为520.9 nm、Zeta电位为-83.78 mV、包封率为77.05%；不同时段脂质体对白纹伊蚊4龄幼虫毒力回归方程分别为y=3.0499x+0.0368（r=0.9771）、y=2.6003x+1.1592（r=0.9600）和y=2.7702x+1.2624（r=0.9765），LC50值分别为42.40、30.00和22.35 μg•mL-1；脂质体透过家蚕和斜纹夜蛾表皮的效果均优于KRP，48 h后通过家蚕和斜纹夜蛾表皮的累积渗透量达24.67和20.83 μg•mm-2，分别是同时间段KRP的3.10和2.35倍。【结论】以该配方制得的KRP脂质体生物活性明显高于KRP，经皮渗透效果也优于KRP。

关键词: KillerRed 蛋白 , 脂质体 , 生物活性 , 经皮渗透

Abstract: 【Objective】The objective of this study is to research the preparation prescription of KillerRed protein liposome (L-KRP) and its physicochemical properties and bioactivity for optimizing the bioefficacy of KRP. 【Method】 L-KRP was prepared by reverse-phase evaporation method, encapsulation efficiency of L-KRP was determined by the fluorescence spectrophotometry, orthogonal experiment was adopted to optimize the prescription using encapsulation efficiency as the evaluation parameter. The appearance, size, Zeta potential, leakage rate and bioactivity of L-KRP were investigated. 【Result】 The dispersed liposome was spherical with the mean diameter of 520.9 nm, the Zeta potential of -83.78 mV and the mean encapsulation efficiency of 77.05%. The toxicity regression equations of L-KRP against the 4th instars of Aedes albopictus were y=3.0499x+0.0368 (r=0.9771), y=2.6003x+1.1592 (r=0.9600) and y=2.7702x+1.2624 (r=0.9765) at different times, with their LC50 were 42.40, 30.00 and 22.35 μg•mL-1, respectively. The permeability of L-KRP was better than KRP through the epidermis of Bombyx mori and Spodoptera litura, the accumulated permeation through B. mori and S. litura were 24.67 and 20.83 μg•mm-2 after 48 hours, with 3.10 and 2.35 times of KRP, respectively.【Conclusion】 All the bioactivity and permeability of L-KRP were higher than KRP.

Key words: KillerRed protein , liposome , bioactivity , transdermal permeability

殷素侠, 廖美德, 徐汉虹. 光敏蛋白脂质体的制备与生物活性[J]. 中国农业科学, 2013, 46(1): 60-68.

YIN Su-Xia, LIAO Mei-De, XU Han-Hong. Preparation of Photosensitive Protein Liposome and Its Bioactivity[J]. Scientia Agricultura Sinica, 2013, 46(1): 60-68.

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