| 细胞内胆固醇含量对血管平滑肌细胞钙化的影响 |
| 投稿时间：2017-09-17 修订日期：2017-10-08 点此下载全文 |
| 引用本文：刘莎,韩雪,郭维康,刁宗礼,刘文虎.细胞内胆固醇含量对血管平滑肌细胞钙化的影响[J].医学研究杂志,2018,47(10):23-27 |
| DOI：
10.11969/j.issn.1673-548X.2018.10.008 |
| 摘要点击次数: 1048 |
| 全文下载次数: 673 |
|
| 基金项目:国家自然科学基金资助项目（81300607，81570660）；北京市科技计划项目（D131100004713003）；北京市医管局青苗计划（QML20160101） |
|
| 中文摘要:目的 探讨细胞内胆固醇含量对血管平滑肌细胞钙化的影响。方法 分别采用生长培养基（磷1.4mmol/L）和钙化培养基（磷3.0mmol/L），培养人脐静脉血管平滑肌细胞，使用甲基酚酞络合铜法检测细胞基质钙、比色法检测碱性磷酸酶活性、荧光酶标法检测细胞内胆固醇含量，茜素红S染色观察细胞基质钙盐沉积。在钙化培养基下，细胞给予不同浓度的胆固醇刺激，观察上述指标的变化。结果 与生长培养基比较，钙化培养基培养细胞3天、7天时均能增加细胞基质钙、碱性磷酸酶活性和细胞内胆固醇含量，组间和组内差异有统计学意义（P<0.05），培养14天时茜素红染色显示大量染色阳性的钙质沉积。在钙化培养基下，分别添加10、15、20、25μmol/L胆固醇刺激细胞72h，呈剂量依赖式地增加细胞基质钙、碱性磷酸酶活性和细胞内胆固醇含量，组间比较差异有统计学意义（P<0.05）。细胞内胆固醇含量与基质钙、ALP活性呈正相关（P<0.05）。结论 细胞内的高胆固醇含量，能加重高磷诱导的血管平滑肌细胞钙化。 |
| 中文关键词:细胞内胆固醇 血管平滑肌细胞 血管钙化 |
| |
| Effect of Intracellular Cholesterol Content on Vascular Smooth Muscle Cells Calcification |
|
|
| Abstract:Objective To investigate the effect of intracellular cholesterol content on vascular smooth muscle cells calcification. Methods With either growth medium (1.4mmol/L Pi)or calcification medium (3.0mmol/L Pi), primary human umbilical vein smooth muscle cells were incubated. Matrix calcium quantitation was analyzed using the o-cresolphthalein complex one method. Alkaline phosphatase activity was assessed colorimetrical. Total cellular cholesterol content was measured using Fluorence microplate reader. Alizarin Red S staining was used to show the matrix calcification morphologically. Under treatment with calcification medium, vascular cells were further incubated with exogenous cholesterol in different concentrations, to observe the changes of above indexes. Results Compared with treatment using growth medium, treatment using calcification medium for 3 days, 7 days increased matrix calcium, alkaline phosphatase activity and intracellular cholesterol level, the difference of intergroup or between-group was statistically significant(P<0.05). Upon treatment of vascular cells with CM for 14 days, abundant mineral deposits could be detected in the extracellular matrix. Addition of 10、15、20、25μmol/L cholesterol contributed to increased further matrix calcification and alkaline phosphatase activity in a dose-dependent manner, and the difference between the groups was statistically significant(P<0.05).Intracellular cholesterol level was positively correlated with levels of matrix calcification and alkaline phosphatase activity(P<0.05). Conclusion High intracellular cholesterol content contributes to phosphate-induced vascular smooth cells calcification. |
| keywords:Intracellular cholesterol Vascular smooth muscle cell Vascular calcification |
| 查看全文 查看/发表评论 下载PDF阅读器 |
