目的 本研究旨在探讨细胞外基质刚度变化对神经干细胞(neural stem cells,NSCs)分化的影响及其作用机制。方法 本研究基于成功构建脊髓损伤大鼠模型,并制备不同刚度(0.7 k Pa、40 k Pa)的聚丙烯酰胺凝胶基底,将大鼠原代NSCs于不同刚度...目的 本研究旨在探讨细胞外基质刚度变化对神经干细胞(neural stem cells,NSCs)分化的影响及其作用机制。方法 本研究基于成功构建脊髓损伤大鼠模型,并制备不同刚度(0.7 k Pa、40 k Pa)的聚丙烯酰胺凝胶基底,将大鼠原代NSCs于不同刚度基底上培养。压电型机械敏感离子通道组件1 (piezo type mechanosensitive ion channel component 1,Piezo1) sh RNA质粒转染NSCs细胞。免疫荧光染色检测神经元标志物双皮质醇(doublecortion,DCX)和星形胶质细胞标志物胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)阳性细胞百分比。免疫组织化学及蛋白质免疫印迹(Western blot)法检测损伤组织及NSCs细胞中Piezo1蛋白的表达水平。结果 与0.7 k Pa基质刚度组相比,40 k Pa基质刚度组中DCX阳性细胞数增加,而GFAP阳性细胞数减少,Piezo1蛋白表达量上升。脊髓损伤大鼠损伤组织Piezo1蛋白表达显著高于空白对照(sham)组。40 k Pa基质刚度条件下沉默Piezo1后,DCX阳性细胞数减少,而GFAP阳性细胞数增加,差异具有统计学意义(P<0.05)。机制研究发现,沉默Piezo1导致IV型胶原及纤连蛋白表达下降。重组纤连蛋白逆转了Piezo1 sh RNA对NSCs分化的影响,即DCX阳性细胞数增加,而GFAP阳性细胞数减少。结论 综上可见,硬基底刚度通过促进Piezo1蛋白表达,上调IV型胶原及纤连蛋白表达,从而调控NSCs细胞分化。本研究为基于生物材料治疗脊髓损伤提供了新的视角。展开更多
The lsolobal displacement reaction between the dianion {M(CO)3[(h5-C5H4) C(O)C6H4C(O)(h5-C5H4)](CO)3M}2- and two molecules of [(m3-C)CO2Et]Co3(CO)9 gave rise to the formation of terephthaloyl(biscyclopentadienyl)-brid...The lsolobal displacement reaction between the dianion {M(CO)3[(h5-C5H4) C(O)C6H4C(O)(h5-C5H4)](CO)3M}2- and two molecules of [(m3-C)CO2Et]Co3(CO)9 gave rise to the formation of terephthaloyl(biscyclopentadienyl)-bridged double cluster complexes [(h5-C5H4)C(O)C6H4C(O)(h5-C5H4)]{[(m3-C)CO2Et]Co2M(CO)8}2 (M = Mo 1, M = W, 2). In the reaction, single cluster complexes[(m3-C)CO2Et]-Co2M(CO)8[h5-C5H4C(O)C6H4CO2CH3] ( M = Mo 3, M = W, 4) were yielded as by-products. All the cluster complexes were characterized by C/H analysis, IR and 1H NMR spectroscopy, and the structure of 3 has been determined by single-crystal X-ray diffraction method. Crystal data: Mr = 750.19, monoclinic, space group P21/c, a = 22.652(5), b = 8.954 (2), c = 14.530(3) ? b = 104.00(3), V = 2859.6(10) 懦, Z = 4, Dc = 1.743 mg/m? F(000) = 1488, m = 1.644 mm-1, the final R = 0.0555 and wR = 0.1353 for 5519 observations with I > 2s(I).展开更多
文摘目的 本研究旨在探讨细胞外基质刚度变化对神经干细胞(neural stem cells,NSCs)分化的影响及其作用机制。方法 本研究基于成功构建脊髓损伤大鼠模型,并制备不同刚度(0.7 k Pa、40 k Pa)的聚丙烯酰胺凝胶基底,将大鼠原代NSCs于不同刚度基底上培养。压电型机械敏感离子通道组件1 (piezo type mechanosensitive ion channel component 1,Piezo1) sh RNA质粒转染NSCs细胞。免疫荧光染色检测神经元标志物双皮质醇(doublecortion,DCX)和星形胶质细胞标志物胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)阳性细胞百分比。免疫组织化学及蛋白质免疫印迹(Western blot)法检测损伤组织及NSCs细胞中Piezo1蛋白的表达水平。结果 与0.7 k Pa基质刚度组相比,40 k Pa基质刚度组中DCX阳性细胞数增加,而GFAP阳性细胞数减少,Piezo1蛋白表达量上升。脊髓损伤大鼠损伤组织Piezo1蛋白表达显著高于空白对照(sham)组。40 k Pa基质刚度条件下沉默Piezo1后,DCX阳性细胞数减少,而GFAP阳性细胞数增加,差异具有统计学意义(P<0.05)。机制研究发现,沉默Piezo1导致IV型胶原及纤连蛋白表达下降。重组纤连蛋白逆转了Piezo1 sh RNA对NSCs分化的影响,即DCX阳性细胞数增加,而GFAP阳性细胞数减少。结论 综上可见,硬基底刚度通过促进Piezo1蛋白表达,上调IV型胶原及纤连蛋白表达,从而调控NSCs细胞分化。本研究为基于生物材料治疗脊髓损伤提供了新的视角。
基金This work is supported by Natural Science Foundation of China
文摘The lsolobal displacement reaction between the dianion {M(CO)3[(h5-C5H4) C(O)C6H4C(O)(h5-C5H4)](CO)3M}2- and two molecules of [(m3-C)CO2Et]Co3(CO)9 gave rise to the formation of terephthaloyl(biscyclopentadienyl)-bridged double cluster complexes [(h5-C5H4)C(O)C6H4C(O)(h5-C5H4)]{[(m3-C)CO2Et]Co2M(CO)8}2 (M = Mo 1, M = W, 2). In the reaction, single cluster complexes[(m3-C)CO2Et]-Co2M(CO)8[h5-C5H4C(O)C6H4CO2CH3] ( M = Mo 3, M = W, 4) were yielded as by-products. All the cluster complexes were characterized by C/H analysis, IR and 1H NMR spectroscopy, and the structure of 3 has been determined by single-crystal X-ray diffraction method. Crystal data: Mr = 750.19, monoclinic, space group P21/c, a = 22.652(5), b = 8.954 (2), c = 14.530(3) ? b = 104.00(3), V = 2859.6(10) 懦, Z = 4, Dc = 1.743 mg/m? F(000) = 1488, m = 1.644 mm-1, the final R = 0.0555 and wR = 0.1353 for 5519 observations with I > 2s(I).
