牛支原体及其脂质相关膜蛋白诱导胎牛肺细胞IL-1β表达的分子机制研究

The expression of IL-1β in embryonic bovine lung cells induced by Mycoplasma bovis and its lipid-associated membrane proteins

为研究牛支原体(M.bovis)及其脂质相关膜蛋白(LAMPs)诱导胎牛肺(EBL)细胞IL-1β表达的分子机制,本研究首先以M.bovis及其LAMPs刺激EBL细胞,利用荧光定量PCR检测细胞因子IL-1β的动态变化;然后将融合表达的细胞p65(Rel A)蛋白和绿色荧光蛋白重组质粒p EGFP-p65转染EBL细胞,以LAMPs刺激EBL细胞,通过激光共聚焦观察p65的亚细胞分布。荧光定量PCR结果显示,M.bovis及其LAMPs能够诱导EBL细胞IL-1β的表达;而且LAMPs作用的最佳剂量为2μg/m L,最佳时间为12 h。同时,激光共聚焦试验检测到LAMPs能够诱导p65进入EBL细胞核中。上述实验结果表明LAMPs能够诱导EBL细胞中p65进入细胞核内,从而激活NF-κB信号通路,诱导IL-1β上调表达。

In order to study the molecular mechanism of the expression of IL-1β in embryonic bovine lung （EBL） cells induced by Mycoplasma boris and its lipid-associated membrane proteins （LAMPs）, the dynamic expression of IL-1β was initially examined by real-time PCR. Then, the recombinant plasmid pEGFP-p65 was transfected into EBL cells stimulated with LAMPs and the subcellular localization of p65 was analyzed using confocal laser microscopy. Results of real-time PCR showed that M.bovis and its LAMPs were able to induce the expression of IL-1β in EBL cells, and the optimum concentration and time of LAMPs were 2 μg/mL and 12 hours, p65 was translocated into the nucleus of EBL cells visualized via confocal laser scanning microscopy. The above results demonstrated that LAMPs were able to induce the translocation of p65 into the nucleus of EBL cell which leads to the activation of NF-κB signaling pathway, upregnlating the expression of IL-1 β.