Desulfovibrio fairfieldensis外膜囊泡的表征及促炎功能研究
首发时间:2021-03-18
摘要:目的:Desulfovibrio fairfieldensis(D. fairfieldensis)普遍存在于人群肠道微生物中,是一种潜在的病原微生物。通过研究其分泌的外膜囊泡(OMVs)揭示其可能的致病作用及机制。方法:超速离心分离浓缩D. fairfieldensis分泌的OMVs,通过测定粒径、透射电镜观察、label-free非靶定量蛋白组组学对OMVs进行表征。利用人结肠上皮细胞(Caco-2)研究OMVs对上皮屏障功能的作用,利用单核巨噬细胞(THP-1-Mφ)研究其诱导炎症因子分泌的作用。结论:比较D. fairfieldensis及其分泌的OMVs蛋白组学检测结果发现,OMVs结构中的蛋白超过99%均在细菌中有表达,但表达量存在差异。将细菌及OMVs共同表达的蛋白进行功能预测,结果显示这些蛋白均能参与生命活动。从D. fairfieldensis分离得到的OMVs会破坏肠上皮细胞紧密连接屏障,并能显著诱导巨噬细胞分泌大量炎症因子并造成其死亡。
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Characterization and Pro-Inflammation function of Desulfovibrio fairfieldensis-derived outer membrane vesicles
Abstract:Object: Desulfovibrio fairfieldensis(D. fairfieldensis) is a kind of pathogenic microbe, which is widely existing in the intestinal of human. The possible pathogenic effects and mechanisms are revealed by investigating the outer membrane vesicles (OMVs) secreted by D. fairfieldensis. Methods: Ultracentrifugation was used to separate and concentrate OMVs. OMVs were characterized by particle size measurement, transmission electron microscopy, and label-free non-target quantitative proteomics. Vesicles were added to cell culture medium for in-vitro studies. Human colonic epithelial cells (Caco-2) were used to study the effect of OMVs on the epithelial barrier function, and monocyte macrophages (THP-1-Mφ) were used to study their role in inducing the secretion of inflammatory factors. Conclusion: Comparing the proteomics of D. fairfieldensis and its OMVs, it is found that more than 99% of the proteins in the OMVs also expressed in bacteria, but the expressions are different. The predictive-function results of the proteins co-expressed showed that these proteins can participate in live events.Co-incubation of the OMVs withTHP-1-Mφ can significantly induce THP-1-Mφ to secrete a large number of inflammatory factors and cause apoptosis, while theintestinal barrier tight junction functioncould be damaged on Caco-2 cells.
Keywords: pathobionts inflammatory outer membrane vesicles Desulfovibrio fairfieldensis
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