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目的探讨雌激素(17β-estradiol)对大鼠骨髓基质干细胞分化的成骨细胞上皮钠离子通道(ENaC)表达和功能的影响。方法采用大鼠骨髓基质干细胞来源的成骨细胞,通过CCK-8试剂盒和AKP试剂盒分别测定雌激素对细胞增殖和分化的影响,通过半定量PCR方法测定雌激素对成骨细胞ENaC和成骨相关基因表达的影响。结果雌激素能促进初级成骨细胞增殖,且较高浓度时(1×10-7,1×10-5mol.L-1)促增殖作用显著(与对照组相比,P<0.05);联用阿米洛利处理后各组细胞增殖速度均有所降低,其中高浓度E2组显著降低(与E2单独处理组相比,P<0.05);雌激素能提高成骨细胞上α-ENaC、γ-ENaC基因的表达水平,且增加趋势与成骨相关基因(Coll-Ia、OC、ALP、ON)mRNA表达的增加趋势一致。结论雌激素促进骨形成的同时也增加ENaC mRNA表达,而阿米洛利(ENaC抑制剂)能阻断雌激素的促成骨作用,说明ENaC可能参与成骨细胞成骨,提示成骨细胞可能存在一个与ENaC相关的调节新途径,为骨代谢的研究提供了一个新思路。
Objective To investigate the effect of 17β-estradiol on the expression and function of osteoblast-derived epithelial sodium channel (ENaC) in rat bone marrow stromal cells. Methods Osteoblasts derived from rat bone marrow stromal stem cells were used to determine the effect of estrogen on the proliferation and differentiation of cells by CCK-8 kit and AKP kit. The effect of estrogen on the expression of ENaC and Effect of bone related gene expression. Results Estrogen could promote the proliferation of primary osteoblasts. When the concentration of estrogen was higher (1 × 10-7, 1 × 10-5mol.L-1), the proliferation of primary osteoblasts was significant (P <0.05 compared with the control group) After treatment with amiloride, the proliferation rate of all groups decreased, especially in high concentration E2 group (P <0.05 compared with E2 alone group); estrogen could increase the expression of α-ENaC on osteoblasts, γ-ENaC gene expression level, and the increasing trend consistent with the increasing tendency of osteogenic genes (Coll-Ia, OC, ALP, ON) mRNA expression. Conclusion Estrogen can promote the formation of bone and also increase the expression of ENaC mRNA, while amiloride (ENaC inhibitor) can block the effect of estrogen on bone formation, indicating that ENaC may be involved in osteoblast osteoblastic, suggesting that osteoblasts may exist A new regulatory pathway related to ENaC provides a new idea for the study of bone metabolism.