目的:观察17β-雌二醇(17β-E2)对大鼠骨髓间充质干细胞(MSCs)成骨分化过程中钙离子(Ca^2+)通道的影响,阐明17β-E2对MSCs促成骨分化的作用机制。方法:采用密度梯度离心法和贴壁筛选法分离出MSCs,连续传代3次,进行成骨细胞的诱导分化。...目的:观察17β-雌二醇(17β-E2)对大鼠骨髓间充质干细胞(MSCs)成骨分化过程中钙离子(Ca^2+)通道的影响,阐明17β-E2对MSCs促成骨分化的作用机制。方法:采用密度梯度离心法和贴壁筛选法分离出MSCs,连续传代3次,进行成骨细胞的诱导分化。将MSCs分为对照组[单纯成骨细胞培养基(OBM)培养]和不同剂量17β-E2组(在OBM中分别添加0.1、1.0、10.0和100.0 pmol·L^-1 17β-E2)。成骨诱导14 d ,各组细胞经Fluo-3/AM染色后,利用激光扫描共聚焦显微镜测定平均荧光强度(MFI),以MFI代表Ca^2+水平。应用全细胞膜片钳技术记录不同条件下的全细胞Ca^2+电流。结果:采用密度梯度离心法和贴壁筛选法成功分离MSCs。MSCs细胞呈成纤维细胞样,核呈椭圆形,细胞核内可见核仁。传代培养的MSCs生长旺盛,保持原代细胞的形态特征。随着17β-E2浓度的增加,各组细胞的Ca^2+水平也逐渐增强,以100.0 pmol·L^-1 17β-E2组最为明显。与对照组比较,10.0和100.0 pmol·L^-1 17β-E2组的MSCs成骨分化过程中Ca^2+水平和Ca^2+电流峰值明显升高( P <0.05或 P <0.01),0.1和1.0pmol·L^-1 17β-E2组Ca^2+水平和Ca^2+电流峰值差异无统计学意义( P >0.05)。结论:密度梯度离心法和贴壁筛选法分离所得细胞为大鼠MSCs。17β-E2通过增强MSCs Ca^2+通道的开放,增强Ca^2+内向电流发挥促进成骨形成作用,并呈剂量依赖性。展开更多
Data of daily mean temperature, precipitation, sunshine hours and tea yield in Xuancheng area from 1979 to 2008 were used to analyze the relationship between lower temperature in spring and tea yield. The results show...Data of daily mean temperature, precipitation, sunshine hours and tea yield in Xuancheng area from 1979 to 2008 were used to analyze the relationship between lower temperature in spring and tea yield. The results showed during 1979 -2008, the per unit area yield of tea in Xu- ancheng area fluctuated greatly in various years. The relative meteorological yield of tea changed greatly during 1979 -2008, and the decrease in tea yield occurred frequently over the past 30 years. Over the past 30 years, the probability of lower temperature in spring was 56.7%, and the probability of serious and extremely serious lower temperature in spring was 43.4%. In each off year with decrease in tea yield, medium or above lower temperature in spring occurred, showing that the impact of lower temperature in spring on tea yield was very obvious.展开更多
文摘目的:观察17β-雌二醇(17β-E2)对大鼠骨髓间充质干细胞(MSCs)成骨分化过程中钙离子(Ca^2+)通道的影响,阐明17β-E2对MSCs促成骨分化的作用机制。方法:采用密度梯度离心法和贴壁筛选法分离出MSCs,连续传代3次,进行成骨细胞的诱导分化。将MSCs分为对照组[单纯成骨细胞培养基(OBM)培养]和不同剂量17β-E2组(在OBM中分别添加0.1、1.0、10.0和100.0 pmol·L^-1 17β-E2)。成骨诱导14 d ,各组细胞经Fluo-3/AM染色后,利用激光扫描共聚焦显微镜测定平均荧光强度(MFI),以MFI代表Ca^2+水平。应用全细胞膜片钳技术记录不同条件下的全细胞Ca^2+电流。结果:采用密度梯度离心法和贴壁筛选法成功分离MSCs。MSCs细胞呈成纤维细胞样,核呈椭圆形,细胞核内可见核仁。传代培养的MSCs生长旺盛,保持原代细胞的形态特征。随着17β-E2浓度的增加,各组细胞的Ca^2+水平也逐渐增强,以100.0 pmol·L^-1 17β-E2组最为明显。与对照组比较,10.0和100.0 pmol·L^-1 17β-E2组的MSCs成骨分化过程中Ca^2+水平和Ca^2+电流峰值明显升高( P <0.05或 P <0.01),0.1和1.0pmol·L^-1 17β-E2组Ca^2+水平和Ca^2+电流峰值差异无统计学意义( P >0.05)。结论:密度梯度离心法和贴壁筛选法分离所得细胞为大鼠MSCs。17β-E2通过增强MSCs Ca^2+通道的开放,增强Ca^2+内向电流发挥促进成骨形成作用,并呈剂量依赖性。
基金Supported by the Research Foundation of Xuancheng Meteorological Bureau in 2013(KY201305)
文摘Data of daily mean temperature, precipitation, sunshine hours and tea yield in Xuancheng area from 1979 to 2008 were used to analyze the relationship between lower temperature in spring and tea yield. The results showed during 1979 -2008, the per unit area yield of tea in Xu- ancheng area fluctuated greatly in various years. The relative meteorological yield of tea changed greatly during 1979 -2008, and the decrease in tea yield occurred frequently over the past 30 years. Over the past 30 years, the probability of lower temperature in spring was 56.7%, and the probability of serious and extremely serious lower temperature in spring was 43.4%. In each off year with decrease in tea yield, medium or above lower temperature in spring occurred, showing that the impact of lower temperature in spring on tea yield was very obvious.
