甲状旁腺激素对大鼠骨髓间充质干细胞钙离子的影响

Effects of parathyroid hormone on calcium ions in rat bone marrow mesenchymal stem cells

背景:甲状旁腺激素可通过靶器官表面的特异性受体激活一系列生理生化反应,调节钙的动态平衡。目的:观察甲状旁腺激素对骨髓间充质干细胞内钙离子的影响,探讨其维持骨髓间充质干细胞体外增殖的机制。方法:将第4代SD大鼠骨髓间充质干细胞分组培养,实验组加入含甲状旁腺激素的培养基,对照组加入常规培养基,培养10 d内,MTT法检测细胞增殖;培养7,14 d,通过激光共聚焦显微镜观察实验组及对照组细胞内钙离子荧光峰值。结果与结论:①细胞增殖:培养第一二天,2组细胞生长活性均滞缓,3 d后进入快速增长期,在第七八天进入平台期,第10天细胞增殖能力有减弱趋势,其中对照组细胞增殖能力低于实验组(P<0.05);②激光共聚焦显微镜观察结果:两组于培养第7天可见胞内呈现较强的钙荧光峰值,随着诱导时间延长,14 d的细胞内钙荧光峰值出现下降,实验组培养7,14 d的细胞内的钙荧光峰值强于对照组(P<0.001),且实验组14 d的细胞内钙荧光峰值与对照组7 d的细胞内钙荧光峰值无差异;③结果表明:甲状旁腺激素可增加骨髓间充质干细胞内钙离子荧光峰值,维持骨髓间充质干细胞增殖功能。

BACKGROUND:Parathyroid hormone can activate a series of physiological and biochemical reactions throughspecific receptors on the surface of target organs,to adjust the dynamic calcium balance.OBJECTIVE:To explore the mechanisms whereby parathyroid hormone maintains in vitro proliferation of bone marrowmesenchymal stem cells(BMSCs),by observing the effects of parathyroid hormone on calcium ions in BMSCs.METHODS:BMSCs at passage4were randomly divided into two groups.Experimental group was treated withparathyroid hormone,and control group was treated with routine culture medium.During10days of culture,MTT assaywas used to detect BMSCs proliferation.At7and14days after treatment,laser scanning confocal microscopy was usedto detect fluorescence intensity of calcium ions in BMSCs in the two groups.RESULTS AND CONCLUSION:(1)At1and2days after culture,the cell viability in the two groups was sluggish.At3days after culture,the cells turned into the rapid growth.At7and8days after culture,the cell growth turned into theplateau.At10days after culture,the cell proliferation ability had a weakening trend,which was lower in the control groupthan in the experimental group(P<0.05).(2)At7days after culture,the fluorescence intensity of intracellular calciumions was strong.As the induction time extended,the fluorescence intensity of intracellular calcium ions was declined at14days after-culture.At7and14days after culture,the fluorescence intensity of intracellular calcium ions in theexperimental group was higher than that in the control group(P<0.001).There was no difference in the fluorescenceintensity of intracellular calcium ions between experimental group at14days and control group at7days.To conclude,parathyroid hormone could increase the fluorescence intensity of calcium ions in BMSCs,and function to maintain theBMSCs proliferation.