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目的本文探讨上皮钠离子通道(ENa C)对破骨细胞功能和活性的影响。方法采用大鼠巨噬细胞集落刺激因子和核转录因子-κB受体活化因子配体诱导大鼠骨髓单核细胞使其分化为破骨细胞。以1.5×104密度接种到12孔板,查随机表分3孔为1组,分为4组:Control组和不同浓度阿米洛利(Ami,ENa C的抑制剂)组。抗酒石酸酸性磷酸酶(TRAP)染色进行阳性破骨细胞鉴定;将破骨细胞和骨片共同培养,测定骨吸收陷窝的数目;用RT-PCR技术分析破骨细胞标志酶基因组织蛋白酶K(CK)的表达。结果不同浓度的Ami处理破骨细胞后,TRAP染色阳性破骨细胞减少,抑制破骨细胞的形成和骨吸收,而且降低破骨细胞特异性基因CK的表达。结论本次实验在细胞水平证明ENa C在破骨细胞上的表达并且调控破骨细胞的分化和骨吸收,说明ENa C可能参与破骨细胞的功能调节,提示破骨细胞可能存在一个与ENa C相关调节的新途径,为骨代谢的研究提供了一个新思路。
Objective To investigate the effect of epithelial sodium channel (ENaC) on the function and activity of osteoclasts. Methods Rat bone marrow mononuclear cells were induced to differentiate into osteoclasts by using rat macrophage colony-stimulating factor and nuclear factor-κB receptor activator ligand. The cells were inoculated into 12-well plates at a density of 1.5 × 10 4. Randomly divided into 3 groups: control group and amiloride (Ami, inhibitor of ENa C) group. Tartrate-resistant acid phosphatase (TRAP) staining. The osteoclasts and bone fragments were co-cultured to determine the number of bone resorption lacuna. The expression of osteoclast marker gene cathepsin K ( CK) expression. Results After treatment with different concentrations of Ami, the number of TRAP-positive osteoclasts was decreased, the formation of osteoclasts and bone resorption were inhibited, and the expression of osteoclast-specific gene CK was also decreased. Conclusions This experiment demonstrates that ENa C is expressed on osteoclasts at the cellular level and regulates osteoclast differentiation and bone resorption, indicating that ENa C may be involved in the functional regulation of osteoclasts, suggesting that there may be a relationship between ENa C and ENa C Relevant regulation of new ways for the study of bone metabolism provides a new idea.