Lipopolysaccharides From Non-Helicobacter pylori Gastric Bacteria Potently Stimulate Interleukin-8 Production in Gastric Epithelial Cells
Lipopolysaccharides From Non-Helicobacter pylori Gastric Bacteria Potently Stimulate Interleukin-8 Production in Gastric Epithelial Cells
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DOI：
10.14309/ctg.0000000000000024
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发表时间：
2019-03
影响因子：
3.6
通讯作者：
Natsumi Miyata;Y. Hayashi;S. Hayashi;Kiichi Sato;Y. Hirai;H. Yamamoto;K. Sugano
中科院分区：
文献类型：
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作者：
Natsumi Miyata;Y. Hayashi;S. Hayashi;Kiichi Sato;Y. Hirai;H. Yamamoto;K. Sugano
背景：慢性幽门螺杆菌（h.p ylori）感染导致胃酸分泌受损，导致非幽门螺杆菌过度生长。胃中的幽门螺杆菌（NHGB）。方法：选择培养基从胃黏膜中分离NHGB，采用生化方法和16S rRNA基因测序对其进行鉴定。采用间接免疫荧光法对人胃组织进行了幽门螺杆菌和亚黄奈瑟菌抗体的研究。胃上皮细胞系与细菌共培养或与NHGB分离的脂多糖共培养，采用酶联免疫吸附法测定培养基中白细胞介素8的释放量。采用免疫荧光显微镜和逆转录聚合酶链反应研究toll样受体（TLR）2、TLR4及其辅助受体髓样分化因子2 （MD2）和CD14在胃细胞中的表达。结果：与幽门螺杆菌共感染的革兰氏阴性菌以嗜血杆菌属、奈瑟菌属、梭杆菌属和细孔菌属为主。来自N. subflava的脂多糖能刺激MKN45细胞中白细胞介素-8的分泌，这种分泌被多粘菌素b预孵育所取消。TLR2、TLR4、CD14和髓样分化因子2在MKN45细胞中表达，尽管它们的表达水平较低。N. subflava在体外粘附于MKN45细胞，并在人胃粘膜中与幽门螺杆菌共定位。结论：我们的数据表明，胃黏膜定殖的亚黄芽孢杆菌参与了慢性幽门螺杆菌胃炎期间的胃炎症。转化影响：NHGB可能使胃炎症长期存在，并加速次氯酸胃的肿瘤进展。
BACKGROUND: Gastric acid secretion is compromised in chronic Helicobacter pylori (H. pylori) infection allowing overgrowth of non-H. pylori gastric bacteria (NHGB) in the stomach. METHODS: NHGB were isolated from gastric mucosa in selective media and further characterized with biochemical methods and 16S rRNA gene sequencing. Human gastric tissues were studied with indirect immunofluorescence with antibodies against H. pylori and Neisseria subflava (N. subflava). Gastric epithelial cell lines were cocultured with bacteria or incubated with lipopolysaccharides isolated from NHGB, and interleukin-8 released in the media was measured by enzyme-linked immunosorbent assay. Expression of Toll-like receptor (TLR)2, TLR4, it's coreceptor myeloid differentiation factor 2 (MD2), and CD14 in gastric cells was investigated by immunofluorescence microscopy and reverse transcriptase-polymerase chain reaction. RESULTS: Haemophilus species, Neisseria species, Fusobacterium species, and Veillonella species were predominant Gram-negative bacteria coinfected with H. pylori. Lipopolysaccharides from N. subflava potently stimulated interleukin-8 secretion in MKN45 cells which was cancelled by preincubation with polymyxin B. TLR2, TLR4, CD14, and myeloid differentiation factor 2 were expressed in MKN45 cells, though their levels of expression were low. N. subflava adhered to MKN45 cells in vitro and colocalized with H. pylori in the human gastric mucosa. CONCLUSIONS: Our data suggest that N. subflava colonized in the gastric mucosa contribute to gastric inflammation during chronic H. pylori gastritis. TRANSLATIONAL IMPACT: NHGB may perpetuate gastric inflammation and accelerate neoplastic progression in the hypochlorhydric stomach.