目的:观察金雀异黄酮通过Ca^(2+)-钙调蛋白依赖性蛋白激酶IV(calmodulin-dependent protein kinase IV,CaMKIV)通路对Aβ_(25-35)诱导海马神经元损伤的保护作用。方法:取24 h内新生SD乳鼠的海马组织,进行神经元的分离纯化和培养,并用免...目的:观察金雀异黄酮通过Ca^(2+)-钙调蛋白依赖性蛋白激酶IV(calmodulin-dependent protein kinase IV,CaMKIV)通路对Aβ_(25-35)诱导海马神经元损伤的保护作用。方法:取24 h内新生SD乳鼠的海马组织,进行神经元的分离纯化和培养,并用免疫荧光染色进行鉴定。神经元细胞随机分为空白对照组、模型组、金雀异黄酮组(50μmol/L)和阳性对照戊酸雌二醇组(10μmol/L),金雀异黄酮组和戊酸雌二醇组预处理3 h后,除空白对照组外,其他各组采用Aβ_(25-35)诱导海马神经元构建细胞损伤模型。利用噻唑蓝法检测细胞存活率,荧光探针检测神经元细胞内Ca^(2+)荧光强度,Western blot检测钙调蛋白(calmodulin,CaM)、钙调蛋白依赖性蛋白激酶激酶(calcium/calmodulin dependent protein kinase kinase,CaMKK)、磷酸化钙调蛋白依赖性蛋白激酶IV(p-calmodulin-dependent protein kinase,p-CaMKIV)和p-Tau蛋白的相对表达量。结果:免疫荧光分析结果显示大鼠海马神经元分离成功。与空白对照组比较,模型组海马神经元细胞存活率极显著下降(P<0.01),细胞Ca^(2+)荧光强度极显著升高(P<0.01),CaM、CaMKK、p-CaMKIV和p-Tau蛋白相对表达量极显著提高(P<0.01);与模型组比较,金雀异黄酮极显著提高了Aβ_(25-35)所致海马神经元损伤模型中细胞的存活率(P<0.01),降低了细胞Ca^(2+)荧光强度(P<0.01),下调了CaM、CaMKK、p-CaMKIV和p-Tau蛋白相对表达量(P<0.01)。结论:金雀异黄酮对Aβ25-35诱导的海马神经元损伤具有明显的神经保护作用,其作用可能是通过Ca^(2+)-CaMKIV通路介导的。展开更多
The bark of Pteroce/tis tatarinowii is a raw material for manufacturing XuanPaper. The effects of Ca^(2+) concentrations on the accumulation of mineral elements in the bark,leaf and root of Pteroceltis tatarinowii wer...The bark of Pteroce/tis tatarinowii is a raw material for manufacturing XuanPaper. The effects of Ca^(2+) concentrations on the accumulation of mineral elements in the bark,leaf and root of Pteroceltis tatarinowii were studied under controlled conditions. The types ofHoagland nutrient solution with three Ca^(2+) concentrations levels (200, 400 and 600 μg·g^(-1))and a control (without Ca^(2+)) were designed to culture Pteroceltis tatarinowii. After 6 months,contents of seven mineral elements including Ca, K, Mg, Mn, Zn, Cu and Na in the root, leaf and barkwere analyzed. The results indicated that Ca accumulations content in the root, leaf and bark hadpositively relation with Ca^(2+) concentrations (200, 400, 600 μg · g^(-1)), and the order of theCa content in the three components was root>leaf>bark. Ca content in the root treated with 600 μg·g^(-1) Ca^(2+) concentrations was 5.5 times as high as that of the control, and about 1.4 times ashigh as that of the root treated in 200 and 400 μg/g Ca^(2+) concentrations respectively. On thecontrary, K and Mg contents in the root, leaf and bark were negatively related to Ca^(2+)concentrations, especially in the bark, and their accumulation trend followed the order ofleaf>root>bark. K content in the bark treated with 600 μg ·g^(-1) Ca^(2+) concentrations was 39.3%of that of the control, and was 79.0% and 91.8% of that of the bark treated with 200 μg ·g^(-1)and 400 μg ·g^(-1) Ca^(2+) concentrations respectively; Mg content in the bark treated with 600μg ·g^(-1) Ca^(2+) concentrations was 23.4% of that of the control, and was 27.1% and 35.4% ofthat of the bark treated with 200 and 400 μg·g^(-1) Ca^(2+) concentrations respectively. Comparedwith the control, the general tendency of Mn, Zn and Cu content decreased with increasing of Ca^(2+)concentrations and their contents were in the order: root>leaf>bark. Based on the results of thisstudy, the experiment has been useful for providing academic bases in improving the bark quality ofPteroceltis tatarinowii on non-limestone soil.展开更多
文摘目的:观察金雀异黄酮通过Ca^(2+)-钙调蛋白依赖性蛋白激酶IV(calmodulin-dependent protein kinase IV,CaMKIV)通路对Aβ_(25-35)诱导海马神经元损伤的保护作用。方法:取24 h内新生SD乳鼠的海马组织,进行神经元的分离纯化和培养,并用免疫荧光染色进行鉴定。神经元细胞随机分为空白对照组、模型组、金雀异黄酮组(50μmol/L)和阳性对照戊酸雌二醇组(10μmol/L),金雀异黄酮组和戊酸雌二醇组预处理3 h后,除空白对照组外,其他各组采用Aβ_(25-35)诱导海马神经元构建细胞损伤模型。利用噻唑蓝法检测细胞存活率,荧光探针检测神经元细胞内Ca^(2+)荧光强度,Western blot检测钙调蛋白(calmodulin,CaM)、钙调蛋白依赖性蛋白激酶激酶(calcium/calmodulin dependent protein kinase kinase,CaMKK)、磷酸化钙调蛋白依赖性蛋白激酶IV(p-calmodulin-dependent protein kinase,p-CaMKIV)和p-Tau蛋白的相对表达量。结果:免疫荧光分析结果显示大鼠海马神经元分离成功。与空白对照组比较,模型组海马神经元细胞存活率极显著下降(P<0.01),细胞Ca^(2+)荧光强度极显著升高(P<0.01),CaM、CaMKK、p-CaMKIV和p-Tau蛋白相对表达量极显著提高(P<0.01);与模型组比较,金雀异黄酮极显著提高了Aβ_(25-35)所致海马神经元损伤模型中细胞的存活率(P<0.01),降低了细胞Ca^(2+)荧光强度(P<0.01),下调了CaM、CaMKK、p-CaMKIV和p-Tau蛋白相对表达量(P<0.01)。结论:金雀异黄酮对Aβ25-35诱导的海马神经元损伤具有明显的神经保护作用,其作用可能是通过Ca^(2+)-CaMKIV通路介导的。
基金This paper is supported by National Natural Science Foundation of China (No. 39970608).
文摘The bark of Pteroce/tis tatarinowii is a raw material for manufacturing XuanPaper. The effects of Ca^(2+) concentrations on the accumulation of mineral elements in the bark,leaf and root of Pteroceltis tatarinowii were studied under controlled conditions. The types ofHoagland nutrient solution with three Ca^(2+) concentrations levels (200, 400 and 600 μg·g^(-1))and a control (without Ca^(2+)) were designed to culture Pteroceltis tatarinowii. After 6 months,contents of seven mineral elements including Ca, K, Mg, Mn, Zn, Cu and Na in the root, leaf and barkwere analyzed. The results indicated that Ca accumulations content in the root, leaf and bark hadpositively relation with Ca^(2+) concentrations (200, 400, 600 μg · g^(-1)), and the order of theCa content in the three components was root>leaf>bark. Ca content in the root treated with 600 μg·g^(-1) Ca^(2+) concentrations was 5.5 times as high as that of the control, and about 1.4 times ashigh as that of the root treated in 200 and 400 μg/g Ca^(2+) concentrations respectively. On thecontrary, K and Mg contents in the root, leaf and bark were negatively related to Ca^(2+)concentrations, especially in the bark, and their accumulation trend followed the order ofleaf>root>bark. K content in the bark treated with 600 μg ·g^(-1) Ca^(2+) concentrations was 39.3%of that of the control, and was 79.0% and 91.8% of that of the bark treated with 200 μg ·g^(-1)and 400 μg ·g^(-1) Ca^(2+) concentrations respectively; Mg content in the bark treated with 600μg ·g^(-1) Ca^(2+) concentrations was 23.4% of that of the control, and was 27.1% and 35.4% ofthat of the bark treated with 200 and 400 μg·g^(-1) Ca^(2+) concentrations respectively. Comparedwith the control, the general tendency of Mn, Zn and Cu content decreased with increasing of Ca^(2+)concentrations and their contents were in the order: root>leaf>bark. Based on the results of thisstudy, the experiment has been useful for providing academic bases in improving the bark quality ofPteroceltis tatarinowii on non-limestone soil.
