MiR-200b attenuates IL-6 production through IKKβ and ZEB1 in human gingival fibroblasts.
MiR-200b attenuates IL-6 production through IKKβ and ZEB1 in human gingival fibroblasts.
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DOI：
10.1007/s00011-018-1192-1
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发表时间：
2018-12
期刊：
影响因子：
--
通讯作者：
Ogata Y
中科院分区：
文献类型：
--
作者：
Matsui S;Zhou L;Nakayama Y;Mezawa M;Kato A;Suzuki N;Tanabe N;Nakayama T;Suzuki Y;Kamio N;Takai H;Ogata Y
微小RNA（miRNAs）在细胞分化、发育、感染、免疫应答、炎症和肿瘤发生等生物学过程中发挥着重要作用。我们先前报道，与未发炎的牙龈相比，发炎牙龈中miR - 200b的表达显著增加。为了阐明miR - 200b在发炎牙龈中的作用，我们分析了miR - 200b对人牙龈成纤维细胞（HGF）中白细胞介素 - 6（IL - 6）表达的影响。
在用白细胞介素 - 1β（IL - 1β；1 ng/ml）或肿瘤坏死因子 - α（TNF - α；10 ng/ml）刺激后，从HGF中提取总RNA和蛋白质，并用miR - 200b表达质粒或miR - 200b抑制剂进行转染。通过实时定量PCR和蛋白质印迹法分析IL - 6、IL - 1β、核因子κB激酶β抑制剂（IKKβ）、锌指E盒结合同源框1（ZEB1）和E - 钙黏蛋白的mRNA和蛋白质水平。
IL - 1β和TNF - α增加了IL - 6的mRNA和蛋白质水平，并且它们在HGF中被miR - 200b过表达显著抑制，而被miR - 200b抑制剂进一步增加。作为miR - 200b靶基因的IKKβ和ZEB1对E - 钙黏蛋白起负调控作用。miR - 200b抑制了HGF中IKKβ和ZEB1的表达，并增加了E - 钙黏蛋白的mRNA和蛋白质水平。
这些结果表明，miR - 200b通过IKKβ和ZEB1减轻牙周组织中的炎症反应。
MicroRNAs (miRNAs) play important roles in biological processes such as cell differentiation, development, infection, immune response, inflammation and tumorigenesis. We previously reported that the expression of miR-200b was significantly increased in inflamed gingiva compared with non-inflamed gingiva. To elucidate the roles of miR-200b in the inflamed gingiva, we have analyzed the effects of miR-200b on the expression of IL-6 in human gingival fibroblasts (HGF). Total RNA and protein were extracted from HGF after stimulation by interleukin-1β (IL-1β; 1 ng/ml) or tumor necrosis factor-α (TNF-α; 10 ng/ml) and transfected with miR-200b expression plasmid or miR-200b inhibitor. IL-6, IL-1β, inhibitor of nuclear factor kappa-B kinaseβ (IKKβ), Zinc-finger E-box-binding homeobox 1 (ZEB1) and E-cadherin mRNA and protein levels were analyzed by real-time PCR and Western blot. IL-1β and TNF-α increased IL-6 mRNA and protein levels, and they were significantly suppressed by miR-200b overexpression, whereas they were further increased by miR-200b inhibitor in HGF. IKKβ and ZEB1 which are target genes of miR-200b negatively regulate E-cadherin. MiR-200b suppressed the expression of IKKβ and ZEB1 and increased E-cadherin mRNA and protein levels in HGF. These results suggest that miR-200b attenuates inflammatory response via IKKβ and ZEB1 in periodontal tissue.
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影响因子：
6.6
作者：
Katsura A;Tamura Y;Hokari S;Harada M;Morikawa M;Sakurai T;Takahashi K;Mizutani A;Nishida J;Yokoyama Y;Morishita Y;Murakami T;Ehata S;Miyazono K;Koinuma D
通讯作者：
Koinuma D
影响因子：
14.9
作者：
Kozomara A;Griffiths-Jones S
通讯作者：
Griffiths-Jones S
影响因子：
3.7
作者：
Chuang TD;Khorram O
通讯作者：
Khorram O
影响因子：
5
作者：
Cao, Yongmei;Liu, Yujing;Li, Yingchuan
通讯作者：
Li, Yingchuan
影响因子：
14.9
作者：
Betel D;Wilson M;Gabow A;Marks DS;Sander C
通讯作者：
Sander C