猪防御素-1的原核表达及抑菌效应分析

doi:10.11843/j.issn.0366-6964.2025.06.027

摘要/Abstract

摘要：

旨在探究原核表达的猪防御素-1(porcine β-defensin-1)的体内外抑菌作用，为临床乳房炎的治疗提供材料和理论依据。本试验通过构建PBD-1的原核表达载体，优化诱导表达条件，过量表达纯化后测定最小抑菌浓度并绘制杀菌动力曲线。分别用E.coli 13-1和S.aureus 4-1构建小鼠乳房炎模型，并用不同浓度的PBD-1进行乳房内灌注治疗试验，通过乳腺组织病理变化及其载菌量观察治疗效果。结果显示，成功构建PBD-1的原核表达载体，重组PBD-1为可溶性表达，表达量为328.6 mg·mL^-1。体外抑菌试验显示，PBD-1对E.coli 13-1、S.aureus 4-1和S.epidermidis 25-1均有抑制作用，且MIC为82.15 μg·mL^-1。乳房内灌注试验结果显示，1×MIC和2×MIC的PBD-1对S.aureus 4-1和E.coli 13-1小鼠乳房炎模型有一定的治疗作用。重组PBD-1对3种细菌均具有体外抑制作用，且对S.aureus 4-1和E.coli 13-1的乳房炎模型具有治疗作用。

关键词: 原核表达, 乳房炎, 猪防御素-1, 体外抑菌, 动物模型

Abstract:

The aim of this study was to investigate the antibacterial activity in vivo and in vitro of PBD-1 over-expressed by prokaryotes, and to provide material and theoretical basis for the treatment of clinical mastitis. In this study, prokaryotic expression vectors of PBD-1 were constructed, the induced expression conditions were optimized, the minimum inhibitory concentration (MIC) was determined after over-expression and purification, and the bactericidal dynamic curve was drawn. The mouse model of mastitis was established with E. coli13-1 and S. aureus 4-1, and the intramammary perfusion therapy was performed with different concentrations of PBD-1, and the therapeutic effect was observed through the pathological changes of breast tissue and its bacterial load. The results showed that the prokaryotic expression vector of PBD-1 was successfully constructed, the recombinant PBD-1 was soluble and yielded of 328.6 mg·mL^-1. The recombinant PBD-1 had antibacterial effect on E. coli 13-1, S. aureus 4-1 and S. epidermidis 25-1, and the MIC was 82.15 μg·mL^-1 in vitro. The results of intramammary perfusion test showed that PBD-1 with 1×MIC and 2×MIC had a certain therapeutic effect on S. aureus 4-1 andE. coli 13-1 mouse mastitis models. Prokaryotic expression of recombinant PBD-1 has inhibitory effects on all three detected bacteriain vitro, and has therapeutic effects on S. aureus4-1 and E. coli 13-1 mastitis models.

Key words: prokaryotic expression, mastitis, PBD-1, antibacterial activity in vitro, animal model

中图分类号:

S859.797

国德洋, 胡慧, 郑雪莉, 姜艳芬. 猪防御素-1的原核表达及抑菌效应分析[J]. 畜牧兽医学报, 2025, 56(6): 2836-2846.

GUO Deyang, HU Hui, ZHENG Xueli, JIANG Yanfen. Prokaryotic Expression and Analysis of Bacteriostatic Effects of Porcine β-defensin-1[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2836-2846.

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组别 Group		攻毒量/(CFU/50 μL) Infection dosage	细菌计数/(CFU·g^-1) Bacteria count
大肠埃希菌 E.coli	低剂量 Low-dose	6.3×10²	1.6×10⁶
	中剂量 Medium-dose	6.3×10³	8.0×10⁸
	高剂量 High-dose	6.3×10⁴	1.6×10⁹
金黄色葡萄球菌 S. aureus	低剂量 Low-dose	1.4×10³	4.7×10⁵
	中剂量 Medium-dose	1.4×10⁴	4.0×10⁶
	高剂量 High-dose	1.4×10⁵	1.5×10⁷
生理盐水 Normal saline		/	＜10
空白对照 Blank control		/	＜10

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