Osorio-Caballero M, Perdigón-Palacio C, García-López G, Flores-Herrera O, Olvera-Sánchez S, Morales-Méndez I, Sosa-González I, Acevedo J F, Guzmán-Grenfell A M, Molina-Hernández A, Díaz N F, Flores-Herrera H
Department of Obstetrics and Gynecology, National Institute of Perinatology "Isidro Espinosa de los Reyes", Montes Urales #800, Col. Lomas de Virreyes cp, 11000 Mexico City, Mexico.
Department of Biochemistry and Molecular Biology, National Institute of Perinatology "Isidro Espinosa de los Reyes", Mexico City, Mexico.
Placenta. 2015 Mar;36(3):262-9. doi: 10.1016/j.placenta.2014.12.011. Epub 2014 Dec 24.
Escherichia coli is recognized as an etiological bacteria associated with chorioamnionitis and the preterm premature rupture of fetal membranes. This pathological condition induces pro-inflammatory cytokines and degradative metalloproteinases, which are considered biological markers secreted in an acute stage of infection. Heat-shock proteins (HSPs) are an important component of the innate immunity response and are found in different pathological conditions. They have not been previously measured in human fetal membranes in response to infectious conditions. We hypothesized that the choriodecidual tissue and amniotic epithelium secreted temporal and differential Hsp-60, Hsp-70, and interleukin (IL)-1β mediated by E. coli infection.
Fetal membranes were mounted in a two-compartment culture system and infected with two passes of live E. coli at different doses (10², 10⁴, 10⁵, and 10⁶ colony-forming units (CFU)/mL) and intervals of incubation (3, 6, and 24 h). The culture medium was collected, and Hsp-60, Hsp-70, and IL-1β were assessed using the enzyme-linked immunosorbent assay (ELISA) method.
After 3 and 6 h of infection, E. coli induced an increase in Hsp-70 secretion in the choriodecidual tissue. However, after 24 h of incubation, Hsp-70 was downregulated and we observed an increase in IL-1β secretion. By contrast, E. coli induced a lower Hsp-60 secretion in the amnion compared to Hsp-70.
Human fetal membranes responded actively to E. coli infection, with an increase in Hsp-70 during the first hours of infection. After 24 h, there was an increase in the liberation of IL-1β.
大肠杆菌被认为是与绒毛膜羊膜炎和胎膜早破相关的病原菌。这种病理状况会诱导促炎细胞因子和降解性金属蛋白酶，它们被视为感染急性期分泌的生物标志物。热休克蛋白（HSPs）是先天免疫反应的重要组成部分，在不同病理状况下均可发现。此前尚未在人类胎膜中检测过其对感染状况的反应。我们推测，大肠杆菌感染可介导绒毛蜕膜组织和羊膜上皮分泌不同时期的热休克蛋白60（Hsp-60）、热休克蛋白70（Hsp-70）和白细胞介素（IL）-1β。
将胎膜置于双室培养系统中，用不同剂量（10²、10⁴、10⁵和10⁶菌落形成单位（CFU）/mL）的活大肠杆菌进行两轮感染，并在不同孵育时间间隔（3、6和24小时）进行培养。收集培养基，采用酶联免疫吸附测定（ELISA）法评估Hsp-60、Hsp-70和IL-1β。
感染3小时和6小时后，大肠杆菌诱导绒毛蜕膜组织中Hsp-70分泌增加。然而，孵育24小时后，Hsp-70表达下调，且我们观察到IL-1β分泌增加。相比之下，与Hsp-70相比，大肠杆菌诱导羊膜中Hsp-60分泌较低。
人类胎膜对大肠杆菌感染有积极反应，在感染的最初几个小时内Hsp-70增加。24小时后，IL-1β释放增加。
