PBD-1修饰的BMSCs移植治疗猪支原体肺炎的实验性研究

To make research and develop a new approach of treating Swine Mycoplasma hyopneumoniae without drug residue and resistance exerts an important influence as well as utility value on the healthy development of swine husbandry. The project is implemented on the basis of the newest theory and experimental study of genetic-modified Bone Mesenchymal Stem Cell, combing with the phenomenon of comparatively positive effect the culture supernatant exerted on Swine Mycoplasma hyopneumoniae in the previous vaccine development,with the innovative application of implanting Bone Mesenchymal Stem Cells genetic-modified with PBD-1 into the experimental swine infected by Swine Mycoplasma hyopneumoniae，to explore the effective of purifying Swine Mycoplasma hyopneumoniae infection, and study the immune effect of PBD-1. The result of the study will have not only provided a brand new effective approach to curbing Swine Mycoplasma hyopneumoniae,but also the scientific reference for the treatment of antibacterial and drug-resistant animal disease and even some human disease.

研究和开发一种新的无药物残留、无耐药性的净化猪支原体肺炎的有效途径对于养猪业的健康发展具有重要的科学意义和实用价值。本项目是基于目前骨髓间充质干细胞体内移植的最新理论和临床研究基础，结合前期疫苗研发过程中发现转PBD-1细胞培养上清液对肺炎支原体有较好抑制效果的现象，创新性地运用转染PBD-1的猪自体骨髓间充质干细胞移植到感染猪肺炎支原体的试验猪体内，探讨PBD-1净化支原体感染的效果并分析相关免疫学效应。本项目研究获得的成果将不仅为猪支原体肺炎的治疗提供一条崭新的有效途径，也为其他抗生素耐药性动物疾病甚至是某些人类疾病的治疗提供相关科学依据。

猪支原体肺炎已经成为当今规模化养猪场的四大疾病杀手之一，给生猪产业带来十分恶劣影响。本项目通过差速贴壁法从细胞群中分离出猪骨髓间充质干细胞的亚型细胞BMSC-S建立细胞系，并进一步通过基因工程技术，建立了稳定转染PBD-1基因的BMSC-S/PBD-1细胞系。细胞生物学特性试验研究表明，猪BMSC-S/PBD-1细胞与BMSC细胞具有类似的干细胞生物学特性。经单个细胞克隆方法筛选，获得9个稳定表达绿色荧光蛋白和PBD-1的BMSC-S/PBD-1细胞克隆株。对转PBD-1猪BMSCs细胞培养上清抑菌效果分析，发现了转PBD-1猪BMSCs培养上清（冻干粉）对肺炎支原体（Mhp）体外生长具有较好抑制效果。将稳定转染PBD-1基因的BMSC-S/PBD-1细胞通过耳缘静脉输注到支原体肺炎的猪模型体内，可以有效治愈猪支原体肺炎疾病。本项目研究获得的成果将不仅为猪支原体肺炎的治疗提供一条崭新的有效途径，也为其他抗生素耐药性动物疾病甚至是某些人类疾病的治疗提供相关科学依据。