卵黄抗体对感染败血症小鼠的保护作用

发布时间：2019-06-07 17:21

【摘要】：败血症是指病原菌通过各种途径侵入血液,并在血液内生长繁殖产生大量毒素,引起严重的全身性感染。目前败血症主要通过抗生素进行治疗,但由于抗生素滥用和抗生素残留带来的耐药性问题已越来越严重。卵黄抗体(Egg Yolk Immunoglobulin, IgY)有高效、安全、不产生耐药性、无毒副作用等特点广泛应用于各种疾病的防控。卵黄抗体被视为一种最具应用前景的抗生素替代品。但卵黄抗体对于非肠道疾病的研究较少,本研究建立小鼠败血症模型,探讨特异性卵黄抗体对小鼠败血症的防治效果。为卵黄抗体对全身性感染疾病的应用研究提供了实验基础,为卵黄抗体作用机制的研究提供了新的动物模型。 本研究以灭活的金黄色葡萄球菌免疫蛋鸡制备特异性卵黄抗体。采用酶联免疫吸附法测定卵黄抗体效价,结果显示抗体最高效价达到1：18000,抗体高效价(1：14000)可持续5周。采用水稀释-两次盐析-超滤-冷冻干燥技术分离纯化卵黄抗体,最终获得抗体纯度达83.4%。体外抑菌实验表明,特异性卵黄抗体能有效抑制金黄色葡萄球菌的生长,在卵黄抗体浓度为5mg/ml时就能起到显著抑菌效果。免疫荧光结果表明特异性卵黄抗体能与金黄色葡萄球菌特异性结合。 用金黄色葡萄球菌通过尾静脉注射建立小鼠败血症模型,通过口服或腹腔注射特异性卵黄抗体对败血症小鼠进行防治,观察小鼠健康状体及死亡率。实验结果显示,通过尾静脉注射4×107CFU的金黄色葡萄球菌可致小鼠患败血症且死亡率为100%,该剂量为小鼠败血症最小致死量。对小鼠攻毒前采用口服灌胃特异性卵黄抗体进行保护,攻毒后持续3天。小鼠出现寒颤、聚堆、竖毛、行动迟缓、眼角有脓状物,不进食、不饮水等不良现象,且与对照组相比小鼠死亡率无显著性变化。而对小鼠攻毒前通过腹腔注射特异性卵黄抗体进行保护,攻毒后持续3天,实验组小鼠死亡率为0%,与对照组死亡率100%相比防治效果极显著(P0.01),采用腹腔注射对小鼠进行保护后小鼠血液中菌数有显著降低,小鼠体重减少量与对照组相比显著降低(P0.01),实验结果说明腹腔注射特异性卵黄抗体可对败血症小鼠起到显著的防治效果,但是口服特异性卵黄抗体对败血症小鼠无明显保护作用。本研究为卵黄抗体应用于全身性感染的防治提供了实验依据。
[Abstract]:Septicemia refers to pathogenic bacteria invading the blood through various ways, and growing and propagating in the blood to produce a large number of toxins, resulting in serious systemic infection. At present, septicemia is mainly treated by antibiotics, but the problem of drug resistance caused by antibiotic abuse and antibiotic residue has become more and more serious. Yolk antibody (Egg Yolk Immunoglobulin, IgY) has the characteristics of high efficiency, safety, no drug resistance, no toxic and side effects and other characteristics widely used in the prevention and control of various diseases. Yolk antibody is regarded as one of the most promising alternatives to antibiotics. However, there are few studies on yolk antibody for non-intestinal diseases. In this study, a mouse septicemia model was established to explore the preventive and therapeutic effect of specific yolk antibody on septicemia in mice. It provides an experimental basis for the application of yolk antibody to systemic infection diseases and a new animal model for the study of the mechanism of yolk antibody. In this study, inactivated Staphylococcus aureus immunized laying hens were used to prepare specific yolk antibodies. The titer of yolk antibody was determined by enzyme-linked immunosorbent assay (Elisa). The results showed that the highest titer of the antibody was 1 鈮，

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