Department of Oral Microbiology and Immunology, College of Dentistry, Dankook University, Cheonan, Republic of Korea.
Department of Oral Microbiology and Immunology, College of Dentistry, Dankook University, Cheonan, Republic of Korea.
Arch Oral Biol. 2021 Apr;124:105062. doi: 10.1016/j.archoralbio.2021.105062. Epub 2021 Jan 23.
The study aimed to examine the diverse bioactivity of lipooligosaccharide extracted from T. denticola cultured in the presence of hemin and quorum-sensing inhibitor.
T. denticola was cultured in the presence or absence hemin or 2(5 H)-furanone, and lipooligosaccharide from T. denticola cultured in various conditions was extracted using an extraction kit. To investigate bioactivity of the lipooligosaccharide, human gingival fibroblasts (HGFs) were treated with the extracted lipooligosaccharide in the presence or absence of Tannerella forsythia lipopolysaccharide. The induction of cytokine expressions was investigated by real-time RT-PCR and ELISA, and the signaling pathway was examined by immunoblotting. To investigate antagonistic mechanisms of the lipooligosaccharide, HGFs were cotreated with fluorescence-labeled T. forsythia lipopolysaccharide and the extracted lipooligosaccharide. Binding of T. forsythia lipopolysaccharide to the cell was analyzed by a flow cytometer.
Lipooligosaccharide induced a low level of cytokine expression at high concentration of hemin or 2(5 H)-furanone. Lipooligosaccharide extracted from T. denticola cultured in higher concentration of hemin and 2(5 H)-furanone had a greater inhibitory effect on induction of cytokine expression by T. forsythia lipopolysaccharide. Further, lipooligosaccharide inhibited the activation of NF-κB and mitogen-activated protein kinase signaling pathways by T. forsythia lipopolysaccharide. Lipooligosaccharide inhibited the binding of T. forsythia lipopolysaccharide to HGFs in the presence of CD14 and LBP.
The characteristics of T. denticola lipooligosaccharide may be altered by bacterial communication and host factors.
本研究旨在研究从培养时有血红素和群体感应抑制剂存在的牙龈卟啉单胞菌中提取的脂寡糖的多种生物活性。
在有或没有血红素或 2(5H)-呋喃酮的情况下培养牙龈卟啉单胞菌，并使用提取试剂盒从不同条件下培养的牙龈卟啉单胞菌中提取脂寡糖。为了研究脂寡糖的生物活性，用人牙龈成纤维细胞（HGFs）在有无坦纳拉福赛思菌脂多糖的情况下用提取的脂寡糖处理。通过实时 RT-PCR 和 ELISA 研究细胞因子表达的诱导情况，并通过免疫印迹法研究信号通路。为了研究脂寡糖的拮抗机制，将荧光标记的坦纳拉福赛思菌脂多糖和提取的脂寡糖共同处理 HGFs。通过流式细胞仪分析 T. forsythia 脂多糖与细胞的结合。
在血红素或 2(5H)-呋喃酮浓度较高时，脂寡糖诱导细胞因子表达水平较低。从培养在较高血红素和 2(5H)-呋喃酮浓度下的牙龈卟啉单胞菌中提取的脂寡糖对坦纳拉福赛思菌脂多糖诱导细胞因子表达的抑制作用更大。此外，脂寡糖抑制了 T. forsythia 脂多糖激活 NF-κB 和丝裂原活化蛋白激酶信号通路。在存在 CD14 和 LBP 的情况下，脂寡糖抑制 T. forsythia 脂多糖与 HGFs 的结合。
T. denticola 脂寡糖的特性可能会因细菌通讯和宿主因素而改变。
