目的探讨α-1,6岩藻糖基转移酶(FUT8)对人胚肺成纤维细胞(MRC-5)增殖、迁移和纤维化的作用及可能机制。方法将24只C57/BL6小鼠随机分为对照组、博莱霉素组、sh-NC组和sh-FUT8组,应用Masson染色观察肺纤维化情况。细胞实验分别设置对照组...目的探讨α-1,6岩藻糖基转移酶(FUT8)对人胚肺成纤维细胞(MRC-5)增殖、迁移和纤维化的作用及可能机制。方法将24只C57/BL6小鼠随机分为对照组、博莱霉素组、sh-NC组和sh-FUT8组,应用Masson染色观察肺纤维化情况。细胞实验分别设置对照组:MRC-5正常培养;TGF-β1组:TGF-β1处理MRC-5;si-NC组:si-NC转染MRC-5后TGF-β1处理;si-FUT8组:siFUT8转染MRC-5后TGF-β1处理;Gal-3组:si-FUT8和pcDNA3.1-Gal转染MRC-5后TGF-β1处理。CCK-8和BrdU方法检测细胞增殖;划痕实验检测细胞迁移;RT-qPCR和Western blot检测α-平滑肌肌动蛋白(α-SM A)、I型胶原蛋白(COLIA1)和细胞外基质纤维连接蛋白(FN)的表达水平。同时,免疫共沉淀检测了FUT8与半乳糖凝集素-3(Gal-3)的作用及沉默FUT8对FAK/Akt信号通路的调节。结果沉默FUT8显著降低博莱霉素诱导的小鼠肺组织细胞外胶原沉积。沉默FUT8抑制TGF-β1介导的细胞增殖(186.81±6.29 vs 118.09±9.48,P<0.05)和迁移。FUT8缺失下调α-SMA、COLIA1和FN的mRNA和蛋白表达水平(P<0.05)。此外,FUT8可直接与Gal-3作用。沉默FUT8下调Gal-3的表达并抑制FAK/Akt信号通路,过表达Gal-3逆转FUT8对细胞增殖、迁移和纤维化的作用(P<0.05)。结论FUT8通过调控Gal-3调控TGF-β1介导的MRC-5细胞增殖、迁移和纤维化,FAK/Akt信号通路可能参与了这个过程。展开更多
目的本研究通过研究维生素A缺乏(vitamin A deficiency,VAD)对初生鼠FUT8(α-1,6fucosyltransferase,α-1,6FUCT)的表达,探讨VAD对初生鼠心脏发育的关系及其在糖分子水平上可能的机制。方法通过美国营养学会的改良配方建立VAD大鼠模型。...目的本研究通过研究维生素A缺乏(vitamin A deficiency,VAD)对初生鼠FUT8(α-1,6fucosyltransferase,α-1,6FUCT)的表达,探讨VAD对初生鼠心脏发育的关系及其在糖分子水平上可能的机制。方法通过美国营养学会的改良配方建立VAD大鼠模型。VAD饲料喂养母鼠及所产仔鼠为实验组(VAD组),正常饲料喂养母鼠及所产仔鼠为对照组,在初生鼠中进行比较两组FUT8表达与心脏发育。通过高效液相色谱检测维生素A含量并比较两组产仔率,利用HE染色观察两组心脏发育,采用反转录聚合酶链反应和小扁豆凝集素点印迹的方法检测两组初生鼠FUT8的表达。结果对照组母鼠产仔率高于VAD组(P<0.001);RT-PCR及LCA凝集素点印迹示VAD组子鼠FUT8的表达量在转录水平和蛋白水平均高于对照组;HE染色发现VAD组子鼠心血管内皮细胞形态变得不规则、分散,而且VAD组心肌细胞胞质变得疏松。结论维生素A缺乏组会引起FUT8表达的升高,影响心肌细胞形态的改变,这说明VAD与心脏发育有关,维生素A可能促进心脏发育。展开更多
Fucosyltransferase 8(Fut8)and core fucosylation play critical roles in regulating various biological processes,including immune response,signal transduction,proteasomal degradation,and energy metabolism.However,the fu...Fucosyltransferase 8(Fut8)and core fucosylation play critical roles in regulating various biological processes,including immune response,signal transduction,proteasomal degradation,and energy metabolism.However,the function and underlying mechanism of Fut8 and core fucosylation in regulating adult neurogenesis remains unknown.We have shown that Fut8 and core fucosylation display dynamic features during the differentiation of adult neural stem/progenitor cells(aNSPCs)and postnatal brain development.Fut8 depletion reduces the proliferation of a NSPCs and inhibits neuronal differentiation of aNSPCs in vitro and in vivo,respectively.Additionally,Fut8 deficiency impairs learning and memory in mice.Mechanistically,Fut8 directly interacts with integrinα6(Itga6),an upstream regulator of the PI3kAkt signaling pathway,and catalyzes core fucosylation of Itga6.Deletion of Fut8 enhances the ubiquitination of Itga6 by promoting the binding of ubiquitin ligase Trim21 to Itga6.Low levels of Itga6 inhibit the activity of the PI3K/Akt signaling pathway.Moreover,the Akt agonist SC79 can rescue neurogenic and behavioral deficits caused by Fut8 deficiency.In summary,our study uncovers an essential function of Fut8 and core fucosylation in regulating adult neurogenesis and sheds light on the underlying mechanisms.展开更多
文摘目的探讨α-1,6岩藻糖基转移酶(FUT8)对人胚肺成纤维细胞(MRC-5)增殖、迁移和纤维化的作用及可能机制。方法将24只C57/BL6小鼠随机分为对照组、博莱霉素组、sh-NC组和sh-FUT8组,应用Masson染色观察肺纤维化情况。细胞实验分别设置对照组:MRC-5正常培养;TGF-β1组:TGF-β1处理MRC-5;si-NC组:si-NC转染MRC-5后TGF-β1处理;si-FUT8组:siFUT8转染MRC-5后TGF-β1处理;Gal-3组:si-FUT8和pcDNA3.1-Gal转染MRC-5后TGF-β1处理。CCK-8和BrdU方法检测细胞增殖;划痕实验检测细胞迁移;RT-qPCR和Western blot检测α-平滑肌肌动蛋白(α-SM A)、I型胶原蛋白(COLIA1)和细胞外基质纤维连接蛋白(FN)的表达水平。同时,免疫共沉淀检测了FUT8与半乳糖凝集素-3(Gal-3)的作用及沉默FUT8对FAK/Akt信号通路的调节。结果沉默FUT8显著降低博莱霉素诱导的小鼠肺组织细胞外胶原沉积。沉默FUT8抑制TGF-β1介导的细胞增殖(186.81±6.29 vs 118.09±9.48,P<0.05)和迁移。FUT8缺失下调α-SMA、COLIA1和FN的mRNA和蛋白表达水平(P<0.05)。此外,FUT8可直接与Gal-3作用。沉默FUT8下调Gal-3的表达并抑制FAK/Akt信号通路,过表达Gal-3逆转FUT8对细胞增殖、迁移和纤维化的作用(P<0.05)。结论FUT8通过调控Gal-3调控TGF-β1介导的MRC-5细胞增殖、迁移和纤维化,FAK/Akt信号通路可能参与了这个过程。
文摘目的本研究通过研究维生素A缺乏(vitamin A deficiency,VAD)对初生鼠FUT8(α-1,6fucosyltransferase,α-1,6FUCT)的表达,探讨VAD对初生鼠心脏发育的关系及其在糖分子水平上可能的机制。方法通过美国营养学会的改良配方建立VAD大鼠模型。VAD饲料喂养母鼠及所产仔鼠为实验组(VAD组),正常饲料喂养母鼠及所产仔鼠为对照组,在初生鼠中进行比较两组FUT8表达与心脏发育。通过高效液相色谱检测维生素A含量并比较两组产仔率,利用HE染色观察两组心脏发育,采用反转录聚合酶链反应和小扁豆凝集素点印迹的方法检测两组初生鼠FUT8的表达。结果对照组母鼠产仔率高于VAD组(P<0.001);RT-PCR及LCA凝集素点印迹示VAD组子鼠FUT8的表达量在转录水平和蛋白水平均高于对照组;HE染色发现VAD组子鼠心血管内皮细胞形态变得不规则、分散,而且VAD组心肌细胞胞质变得疏松。结论维生素A缺乏组会引起FUT8表达的升高,影响心肌细胞形态的改变,这说明VAD与心脏发育有关,维生素A可能促进心脏发育。
基金supported in part by the National Natural Science Foundation of China(92049108,82371182)Central Guiding Fund for Local Science and Technology Development Projects(2023ZY1058)the National Key Research and Development Program of China(2017YFE0196600)。
文摘Fucosyltransferase 8(Fut8)and core fucosylation play critical roles in regulating various biological processes,including immune response,signal transduction,proteasomal degradation,and energy metabolism.However,the function and underlying mechanism of Fut8 and core fucosylation in regulating adult neurogenesis remains unknown.We have shown that Fut8 and core fucosylation display dynamic features during the differentiation of adult neural stem/progenitor cells(aNSPCs)and postnatal brain development.Fut8 depletion reduces the proliferation of a NSPCs and inhibits neuronal differentiation of aNSPCs in vitro and in vivo,respectively.Additionally,Fut8 deficiency impairs learning and memory in mice.Mechanistically,Fut8 directly interacts with integrinα6(Itga6),an upstream regulator of the PI3kAkt signaling pathway,and catalyzes core fucosylation of Itga6.Deletion of Fut8 enhances the ubiquitination of Itga6 by promoting the binding of ubiquitin ligase Trim21 to Itga6.Low levels of Itga6 inhibit the activity of the PI3K/Akt signaling pathway.Moreover,the Akt agonist SC79 can rescue neurogenic and behavioral deficits caused by Fut8 deficiency.In summary,our study uncovers an essential function of Fut8 and core fucosylation in regulating adult neurogenesis and sheds light on the underlying mechanisms.
