[关键词]
[摘要]
目的 观察不同剂量的龟板提取物对骨髓间充质干细胞中分化抑制蛋白1(inhibitor of differentiation 1,Id1)的作用。方法 将构建成功的PGL3-Id1启动子用磷酸钙共沉淀法转染大鼠骨髓间充质干细胞,龟板提取物分别作用于转染后的骨髓间充质干细胞12、24、36 h,选取作用最为明显的36 h,每组分别加入0、1、3、30、100 μg/mL龟板提取物,药物作用36 h后收集细胞分别应用萤光素酶报告基因系统、RT-PCR和Western blotting检测Id1的表达。结果 早期、小剂量龟板提取物对Id1的影响不大;随着时间和剂量的增加,Id1的表达水平也逐渐升高。结论 龟板提取物促进了骨髓间充质干细胞中Id1的表达,剂量越大,作用时间越长,促进作用越明显。
[Key word]
[Abstract]
Objective To observe the effect of different dosages of Plastrum Testudinis extracts (PTE) on the expression of inhibitor of differentiation 1 (Id1) in bone marrow derived mesenchymal stem cells (MSCs). Methods The PGL3-Id1 promoter was established and the MSCs were transfected with PGL3-Id1 promoter by calcium phosphate co-precipitation method. The transfected cells were treated with PTE for 12, 24, and 36 h, respectively. Different dosages (0, 1, 3, 30, and 100 μg/mL) of PTE were added into samples of 36 h group. After 36 h treatment, the cells were collected and luciferase activity measurement, RT-PCR, and Western blotting methods were used to detect the expression of Id1. Results The activity of short-term and low dose PTE was not substantial, along with the doses or time increasing, the expression of Id1 in bone marrow derived MSCs increased. Conclusion PTE increases the expression of Id1 in MSCs, and the activity is more significant with the dose and time increases.
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[基金项目]
国家自然科学基金资助项目(30772861);肇庆市科技创新计划项目(2013E181)