Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts.
Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts.
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岩藻糖基转移酶8的过表达逆转了高剂量地塞米松对MC3T3-E1前成骨细胞成骨反应的抑制作用
作者：
Wu Z;Lin T;Kang P;Zhuang Z;Wang H;He W;Wei Q;Li Z
FUT8催化的核心聚焦是TGF-β与TGF-β受体结合的必要条件。采用TGF-β1间接结合法评价TGF-β1与TGFBRs的结合能力，采用茜素红和碱性磷酸酶染色检测成骨分化和矿化能力，采用western blot和定量RT-PCR检测成骨相关蛋白和基因的差异表达。质粒介导的功能获得研究。采用凝集素-印迹法和LCA激光共聚焦法检测核心聚焦修饰程度。我们的研究结果显示，与载体处理相比，高剂量（10−6和10−5 M）地塞米松显著抑制MC3T3-E1细胞的增殖、成骨分化和FUT8 mRNA表达，同时促进脂肪生成相关基因mRNA表达，提示高剂量地塞米松抑制成骨作用可能与下调FUT8有关。高剂量地塞米松作用下，FUT8过表达显著促进MC3T3-E1细胞成骨分化，激活TGF-β/Smad信号，提示FUT8逆转了高剂量地塞米松对成骨的抑制作用。此外，凝集素荧光染色和印迹显示，过表达FUT8显著逆转了高剂量地塞米松对TGFBR1和TGFBR2核心聚焦的抑制作用。此外，间接TGF-β1结合实验显示，在高剂量地塞米松存在下，过表达FUT8显著促进TGF-β1与MC3T3-E1细胞TGFBRs的结合。综上所述，这些结果表明FUT8的过表达有助于抵消地塞米松对TGF-β信号传导和成骨的抑制作用。
Core fucosylation catalyzed by FUT8 is essential for TGF-β binding to TGF-β receptors. Indirect TGF-β1 binding assay was used to evaluate the ability of TGF-β1 to bind to TGFBRs, Alizarin red and alkaline phosphatase staining were used to detect osteogenic differentiation and mineralization ability , western blot and quantitative RT-PCR were used to measure the differential expression of osteogenesis-related proteins and genes. Plasmid-mediated gain-of-function study. The scale of core fucosylation modification was detected by Lectin-blot and LCA laser confocal. Our results showed that compared with vehicle treatment, high-dose (10−6 and 10−5 M) dexamethasone significantly inhibited cell proliferation, osteogenic differentiation, and FUT8 mRNA expression while promoting mRNA expression of adipogenesis-related genes in MC3T3-E1 cells, suggesting that downregulation of FUT8 is involved in the inhibitory effect of high-dose dexamethasone on osteogenesis. Overexpression of FUT8 significantly promoted osteogenic differentiation and activated TGF-β/Smad signaling in MC3T3-E1 cells in the presence of high-dose dexamethasone, suggesting that FUT8 reverses the inhibitory effect of high-dose dexamethasone on osteogenesis. In addition, lectin fluorescent staining and blotting showed that overexpression of FUT8 significantly reversed the inhibitory effects of high-dose dexamethasone on core fucosylation of TGFBR1 and TGFBR2. Furthermore, indirect TGF-β1 binding assay showed that overexpression of FUT8 remarkably promoted TGF-β1 binding to TGFBRs in MC3T3-E1 cells in the presence of high-dose dexamethasone. Taken together, these results suggest that overexpression of FUT8 facilitates counteracting the inhibitory effect of dexamethasone on TGF-β signaling and osteogenesis.
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DOI：
10.1097/med.0000000000000368
发表时间：
2017-12
期刊：
Current opinion in endocrinology, diabetes, and obesity
影响因子：
--
作者：
Hsu E;Nanes M
通讯作者：
Nanes M
通过糖皮质激素受体和AP-1串扰来调节人骨髓基质细胞增殖和分化能力。
影响因子：
6.2
作者：
Carcamo-Orive, Ivan;Gaztelumendi, Ainhoa;Delgado, Jesus;Tejados, Naiara;Dorronsoro, Akaitz;Fernandez-Rueda, Jon;Pennington, Daniel J.;Trigueros, Cesar
通讯作者：
Trigueros, Cesar
影响因子：
3.5
作者：
Imai-Nishiya H;Mori K;Inoue M;Wakitani M;Iida S;Shitara K;Satoh M
通讯作者：
Satoh M
转化生长因子 Beta 受去神经小鼠骨中糖皮质激素依赖性机制的调节。
影响因子：
4.6
作者：
Li Y;Jie L;Tian AY;Zhong S;Tian MY;Zhong Y;Wang Y;Li H;Li J;Sun X;Du H
通讯作者：
Du H
肾间质纤维化中周细胞-肌成纤维细胞转变的新机制：核心岩藻糖基化调控
影响因子：
4.6
作者：
Wang N;Deng Y;Liu A;Shen N;Wang W;Du X;Tang Q;Li S;Odeh Z;Wu T;Lin H
通讯作者：
Lin H