牛支原体脂质相关膜蛋白（LAMPs）诱导胎牛肺细胞IL-1β释放的分子机制研究

Since 2008, the first report about pneumonia caused by Mycoplasma bovis (M.bovis) in China, M.bovis has been an increasingly important factor that threatens the cattle industry in our country. Although great efforts have been made to understand the molecular basis of M.bovis infection, the cellular response to M.bovis infection is still largely unknown. Our previously study indicated that lipid-associated membrane proteins (LAMPs) of M.bovis induce embryonic bovine lung (EBL) cells to produce IL-1β by real-time PCR.. In this study, we further investigated the mechanisms underlying LAMPs inducing EBL cells to produce IL-1β by high throughput cDNA microarray. The differentially expressed (DE) genes were identified in the EBL cells by using the MAS 3.0 software which was based on the Kyoto Encyclopedia of Genes and Genomes database. The interactions between proteins encoded by DE genes were analyzed by STRING. Based on the analysis of the DE genes, the candidate genes which were involved in NF-κB and MAPK signaling pathway were selected for the next functional study. The EGFP, overexpression vector and RNA interference molecules targeting the candidate genes were constructed. The EBL cells were used as cell model to analysis the functions of the DE genes in the IL-1β production induced by M. bovis LAMPs.

自2008年在我国首次报到牛支原体肺炎以来，该病日益成为危害我国养牛业的重要因素。但是关于牛支原体的致病性及其致病机制尚不十分清楚。本实验室前期的研究证实牛支原体脂质相关膜蛋白（LAMPs）能够诱导胎牛肺细胞（EBL）产生IL-1β等细胞因子。. 为阐明牛支原体脂质相关膜蛋白（LAMPs）诱导EBL细胞产生IL-1β的分子机制。本研究拟采用高通量的基因芯片技术并结合MAS 3.0和STRING系统来鉴定牛支原体脂质相关膜蛋白（LAMPs）诱导EBL细胞产生的差异表达基因并对其所处的生物进程、信号通路和相互作用进行分析。同时，挑选出处于NF-κB和MAPK信号通路中差异表达的基因。通过构建差异表达基因的绿色荧光融合质粒，真核表达质粒和RNA干扰分子，研究其在牛支原体脂质相关膜蛋白（LAMPs）诱导EBL细胞IL-1β产生方面的作用。