BMSCs条件培养基改善地塞米松对成骨细胞成骨功能抑制效应的研究

发布时间：2018-07-03 07:53

  本文选题：成骨细胞 + BMSCs　； 参考：《中国修复重建外科杂志》2016年06期

【摘要】：目的探讨BMSCs的旁分泌作用对地塞米松所致成骨细胞成骨功能抑制作用的影响。方法首先通过培养小鼠BMSCs 24 h制备无血清条件培养基备用。将小鼠MC3T3-E1细胞株复苏并传至第3代用于实验,分为4组:A组为对照组;B组于细胞中添加1μmol/L地塞米松;C组于细胞中按1∶1比例添加1μmol/L地塞米松和BMSCs条件培养基;D组仅添加BMSCs条件培养基。培养24 h后收集细胞测定ALP含量,Western blot测定细胞内RUNX2、骨钙素(Osteocalcin)蛋白表达,实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-q PCR)测定细胞内α1-Ⅰ型胶原(collagen typeⅠ-α1,COL1A1)、RUNX2、ALP、Osteocalcin基因表达;21 d后行茜素红染色观察钙结节形成情况。结果培养24 h后,B、C、D组ALP含量均显著低于A组,B组低于C、D组(P0.05),C、D组间差异无统计学意义(P0.05)。Western blot检测示,B组RUNX2蛋白相对表达量显著低于A、C、D组(P0.05),A、C、D组间比较差异无统计学意义(P0.05);B组Osteocalcin蛋白相对表达量均显著低于A、C、D组,A、C组低于D组(P0.05),A、C组间差异无统计学意义(P0.05)。RT-q PCR检测示,B、C、D组RUNX2、Osteocalcin、COL1A1、ALP基因相对表达量均显著低于A组(P0.05);D组RUNX2、Osteocalcin、ALP基因相对表达量均显著高于B、C组,C组显著高于B组(P0.05);D组COL1A1基因相对表达量显著高于B组(P0.05),B、C组间及C、D组间比较差异均无统计学意义(P0.05)。培养6 d时A组细胞死亡;21 d时B、C、D组均可见钙结节阳性染色,且逐渐增强。结论小鼠BMSCs条件培养基能改善地塞米松对成骨细胞成骨功能的抑制效应。
[Abstract]:Objective to investigate the effect of paracrine of BMSCs on osteogenic inhibition of osteoblasts induced by dexamethasone. Methods BMSCs were cultured for 24 h to prepare serum-free conditioned medium. The mouse MC3T3-E1 cell line was resuscitated and transferred to the third passage. Group B added 1 渭 mol / L dexamethasone to the cells of group B and group D added 1 渭 mol / L dexamethasone and BMSCs to the conditioned medium at 1:1. After 24 hours of culture, the content of ALP and the expression of osteocalcin protein were determined by Western blot. The intracellular 伪 1- 鈪，

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