动物实验,上皮钠离子通道 z-bo 2020-08-26 486

　　Objective: To investigate the effect of the epithelial sodium ion channel (ENaC) on the function and activity of osteoclasts.

　　Method: Using rat macrophage colony stimulating factor and nuclear transcription factor-κB receptor activator ligand to differentiate rat bone marrow mononuclear cells into osteoclasts. Inoculate 12-well plates at a density of 1.5 x 104. Check the random table and divide the 3 holes into 1 group and 4 groups. The concentration of amiloride (Ami, ENaC inhibitor) group is different from the control group. Tartrate-resistant acid phosphatase (TRAP) staining was used to identify osteoclasts positive; osteoclasts and bone fragments were co-cultured to determine the number of bone resorption vesicles; RT-PCR osteoclast marker enzyme gene cathepsin K (CK) was used expression.

　　Result: After treating osteoclasts with different concentrations of Ami, TRAP staining-positive osteoclasts decreased, osteoclast formation and bone resorption were inhibited, and the expression of osteoclast-specific gene CK decreased.

　　Conclusion: This experiment proves that ENaC is expressed at the cellular level in osteoclasts and regulates osteoclast differentiation and bone resorption. This indicates that ENaC may be involved in the functional regulation of osteoclasts, and that osteoclasts are related to ENaC.