左归丸含药血清对大鼠骨髓间充质干细胞骨向分化中碱性磷酸酶含量的影响

Effect of Zuogui Pill Medicated Serum on Level of Alkaline Phosphatase During Differentiation from Rat Marrow Mesenchymal Stem Cells to Osteoblasts

摘要:

目的: 研究左归丸含药血清对间充质干细胞骨向分化过程中碱性磷酸酶含量变化的影响。 方法: 采用全骨髓贴壁法培养间充质干细胞,连续进行3次差速贴壁法纯化间充质干细胞。常规诱导组以β-甘油磷酸钠、地塞米松、维生素C诱导间充质干细胞骨向分化,实验组在常规诱导组基础上加以左归丸含药血清。使用茜素红染色钙结节、碱性磷酸酶染色鉴定诱导成功。分别在0,7,14,21 d用比色法检测细胞中碱性磷酸酶含量。 结果: 全骨髓贴壁法和差速贴壁法联合使用可得到较为均一间充质干细胞。用比色法测得碱性磷酸酶含量随着诱导时间延长而增加。 结论: 大鼠间充质干细胞经药物诱导后可以分化为成骨细胞。左归丸含药血清能够促使诱导过程中碱性磷酸酶含量表达增加。

Abstract:

Objective: To investigate the change of ALPase during the differentiation from rat marrow mesenchymal stem cells to osteoblasts induced by Zuogui Pill medicated serum. Method: The method of adherent culture was used to culture marrow mesenchymal stem cells and then consecutively to purify mesenchymal stem cells. In conventional induction group, β-glycerophosphate, dexamethasone and vitamin C was used to induce mesenchymal stem cell to differentiate into osteoblasts. In the experimental group, on the basis of conventional induction Zuogui Pill was applied to get the medicated serum. Alizarin red staining was used for showing calcium nodules and alkaline phosphatase staining was used to identify the successful induction. On the 0, 7^th, 14^th and 21^st day a colorimetric assay of alkaline phosphatase was carried out respectively. Result: Whole bone marrow adherent method combined with differential adhesion method could get the purified mesenchymal stem cells. Alkaline phosphatase measured by colorimetry increased with the prolongation of induction, and the induction by the medicated serum was more significant. Conclousin:Rat mesenchymal stem cells could differentiate into osteoblasts when induced by drugs. Zuogui Pill medicated serum could promote the express of alkaline phosphatase.