负载牛干扰素α/γ聚合物纳米颗粒的制备及其抗病毒活性评价  

Preparation of polymer nanoparticles loaded with bovine interferonα/γand evaluation of their antiviral activity

作　　者：冯晶晶 刘国锰 雷白时 袁万哲[1] 张武超 冯小宇 赵款 FENG Jingjing;LIU Guomeng;LEI Baishi;YUAN Wanzhe;ZHANG Wuchao;FENG Xiaoyu;ZHAO Kuan(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071000,China;Beijing Center for Animal Disease Control and Prevention,Beijing 102600,China)

机构地区：[1]河北农业大学动物医学院,河北保定071000 [2]北京市动物疫病预防控制中心,北京102629

出　　处：《河北农业大学学报》2025年第2期70-79,共10页Journal of Hebei Agricultural University

基　　金：河北省自然科学基金生物农业联合基金重点项目(C2022204249);农业农村部兽用生物制品工程技术重点实验室开放课题(SYKF(23)001);国家兽用生物制品工程技术研究中心开放课题(GTKF(24)006).

摘　　要：由牛病毒性腹泻病毒(BVDV)感染引起的犊牛腹泻对养牛业造成巨大经济损失,为研制有效的防治药物,本研究利用原核表达系统对牛干扰素α(BoIFNα)和牛干扰素γ(BoIFNγ)进行融合表达,成功表达并纯化了重组牛干扰素融合蛋白(rBoIFNα/γ)。体外试验表明,rBoIFNα/γ具有低毒性和高抗病毒作用,抗BVDV活性达9292 U/mL;为进一步提高抗病毒效果和药物作用时间,通过聚乙二醇-聚己内酯(PEG-PCL)将其包裹形成聚合物纳米颗粒(PEG-PCL-rBoIFNα/γNPs),并对其体内外活性进行评估。结果表明,PEG-PCL-rBoIFNα/γNPs为直径约169.09 nm的圆球状颗粒,对细胞无明显毒性作用,在生物液体中持续释放96 h。体内试验表明,注射PEG-PCL-rBoIFNα/γNPs能有效缓解小鼠症状和组织损伤,降低病毒血症水平和组织病毒载量,抑制炎症因子的表达,本研究结果为BVDV的预防和治疗提供了新思路。Calf diarrhea caused by bovine viral diarrhea virus(BVDV)infection causes great economic losses to the cattle industry.In order to develop effective preventive and curative drugs,this study utilized a prokaryotic expression system for the fusion expression of bovine interferonα(BoIFNα)and bovine interferonγ(BoIFNγ),and recombinant bovine interferon fusion protein(rBoIFNα/γ)was successfully expressed and purified.In vitro experiments showed that rBoIFNα/γhad low toxicity and high antiviral effects,with anti-BVDV activity up to 9292 U/mL.To further improve the antiviral effect and drug duration of action,polymer nanoparticles(PEG-PCL-rBoIFNα/γNPs)were formed by encapsulating them with poly(ethylene glycol)-poly(caprolactone)(PEG-PCL)and evaluated for in vivo and in vitro activities.The results showed that the PEG-PCL-rBoIFNα/γNPs were spherical particles with a diameter of around 169.09 nm,which had no significant toxic effects on cells and were released continuously for 96 h in biological fluids.In vivo experiments showed that injection of PEG-PCL-rBoIFNα/γNPs could effectively alleviate the symptoms and tissue damage in mice,reduce the viremia level and tissue viral load,and inhibit the expression of inflammatory factors.The findings provide new ideas for the prevention and treatment of BVDV.

关 键 词：牛干扰素 融合蛋白 聚合物纳米颗粒 抗病毒 牛病毒性腹泻病毒 

分 类 号：S855.3[农业科学—临床兽医学]