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目的:用碱性磷酸酶染色及碱性磷酸酶活性分析的方法,研究孕酮对人牙周膜成纤维细胞(hPDLCs)中碱性磷酸酶(Alkaline phosphatase,ALP)表达的影响。方法:原代培养hPDLCs,取第4～6代细胞用于实验。分为空白对照组,孕酮组,抑制剂组。检测普通培养液和成骨诱导培养液中各组细胞在孕酮作用后ALP染色的变化。用碱性磷酸酶活性分析法检测不同浓度的孕酮对hPDLCs中ALP表达的影响。结果:孕酮能够促进hPDLCs中ALP染色阳性面积的表达,且能够提高hPDLCs中ALP的活性,具有时间及剂量依赖性,抑制剂组各指标减小。结论:孕酮能够上调hPDLCs中碱性磷酸酶的表达。
OBJECTIVE: To study the effects of progesterone on the expression of alkaline phosphatase (ALP) in human periodontal ligament fibroblasts (hPDLCs) by alkaline phosphatase staining and alkaline phosphatase activity assay. Methods: Primary culture hPDLCs, take the 4th to 6th generation cells for experiment. Divided into blank control group, progesterone group, inhibitor group. The changes of ALP staining in progesterone treated cells in normal and osteogenic medium were detected. Alkaline phosphatase activity assay was used to detect the effect of different concentrations of progesterone on the expression of ALP in hPDLCs. Results: Progesterone can promote the expression of ALP-positive area in hPDLCs and increase the activity of ALP in hPDLCs in a time-and dose-dependent manner. Conclusion: Progesterone can up-regulate the expression of alkaline phosphatase in hPDLCs.