金黄色葡萄球菌噬菌体脂质体的制备及胞内抗菌活性

doi:10.11843/j.issn.0366-6964.2025.09.041

摘要/Abstract

摘要： 本试验旨在探讨脂质体作为载体，包封递送噬菌体入胞及噬菌体脂质体对牛乳腺上皮细胞内金黄色葡萄球菌的抗菌作用。本试验采用薄膜水合法制备脂质体，用于包封噬菌体SDQ制备噬菌体脂质体，并测定其酸碱稳定性和热稳定性。使用激光粒度仪测定噬菌体脂质体粒径大小、多分散系数和Zeta电位，结合透射电镜观察噬菌体脂质体结构特征。采用CCK-8法测定噬菌体脂质体对MAC-T细胞活力的影响。通过平板计数法测定噬菌体脂质体入胞效率和胞内杀菌活性。最终，选择大豆卵磷脂、胆固醇、吐温80、十八胺以8∶2∶1∶0.5比例制备脂质体，包封噬菌体SDQ后制备噬菌体脂质体(Lip-p-7)。Lip-p-7包封率大于50%，在温度(4~60 ℃)和pH值(2~11)范围内活性稳定。Lip-p-7平均粒径为1 262.5 nm，Zeta电位从中性转向带负电荷，表明脂质体成功包封噬菌体SDQ；多分散系数(polydispersity index，PDI)小于0.7，表明其均匀分散。制备后的Lip-p-7储存于4 ℃条件下，可在90 d内保持完整活性，具有良好的稳定性。脂质体可有效提升噬菌体SDQ入胞数量，降低MAC-T细胞内金黄色葡萄球菌数量。综上，本试验为脂质体作为载体递送噬菌体入胞，为治疗胞内细菌感染提供参考。

关键词: 脂质体, 噬菌体, 金黄色葡萄球菌, 牛乳腺上皮细胞, 胞内抗菌活性

Abstract: The aim of this study was to investigate the use of liposomes as a carrier to deliver phage into the cell and the antibacterial effect of phage liposomes on Staphylococcus aureus in bovine mammary epithelial cells. In this study, liposome-encapsulated phage SDQ was prepared using the thin-film hydration method. The acid-base stability and thermal stability of the phage liposomes were determined using the double-layer plate method. The particle size, polydispersity coefficient, and Zeta potential of phage liposomes were measured using a laser particle size analyzer, while the structural characteristics of the phage liposomes were observed through transmission electron microscopy. The effect of phage liposomes on the viability of MAC-T cells was determined using the CCK-8 assay. The efficiency of cell entry and the intracellular and bactericidal activity of phage liposomes were measured using the plate counting method. The results showed that liposomes made from soybean lecithin, cholesterol, Tween 80, and octadecylamine were prepared in a ratio of 8:2:1:0.5, and phage liposomes (Lip-p-7) were created by encapsulating phage SDQ. The encapsulation rate of Lip-p-7 was greater than 50%, and its activity is stable in the range of temperature (4-60 ℃) and pH value (2-11). The average particle size of Lip-p-7 was 1 262.5 nm, and the shift of Zeta potential from neutral to negatively charged signified the successful encapsulation of phage SDQ by the liposome. The polydispersity index (PDI) was less than 0.7, suggesting a uniform dispersion. When stored at 4 ℃, the prepared Lip-p-7 could maintain its intact activity for up to 90 days, demonstrating good stability. Liposomes can effectively increase the number of phage SDQ entering cells and decrease the number of Staphylococcus aureus in MAC-T cells. In conclusion, this study provides a reference for the treatment of intracellular bacterial infections by using liposomes as a carrier to deliver bacteriophages into the cells.

Key words: liposomes, bacteriophage, Staphylococcus aureus, MAC-T cells, intracellular antibacterial activity

中图分类号:

Q939.48

杨书博, 苑庆欣, 陈麒百, 王培, 高东阳, 李鹤, 宋军. 金黄色葡萄球菌噬菌体脂质体的制备及胞内抗菌活性[J]. 畜牧兽医学报, 2025, 56(9): 4638-4645.

YANG Shubo, YUAN Qingxin, CHEN Qibai, WANG Pei, GAO Dongyang, LI He, SONG Jun. Preparation and Intracellular Antibacterial Activity of Liposomes Encapsulating Staphylococcus aureus Bacteriophage[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4638-4645.

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