新橙皮苷对破骨细胞分化影响的实验研究

Experimental study on effect of neohesperidin on osteoclasts differentiation

目的探讨新橙皮苷对破骨细胞凋亡率、细胞活力以及分化的影响。方法分别在RAW264. 7细胞及小鼠骨髓巨噬细胞中加入核因子-κB受体活化因子配体(RANKL)以及新橙皮苷进行培养分化,对RAW264. 7细胞进行细胞凋亡实验,对小鼠骨髓巨噬细胞进行细胞活力实验以及破骨细胞分化的时间依赖性实验。结果随着新橙皮苷浓度的增加,各组健康细胞率均保持于90%以上的高水平,而坏死率和凋亡率均维持在低水平。以0μM组为对照,各浓度组药物对细胞活力均无影响(P> 0. 05)。第1、3、5、7天加入10μM的新橙皮苷均对破骨细胞形成有抑制作用(P <0. 01),其中第5天加入的新橙皮苷对破骨细胞形成的抑制作用最为显著(P <0. 001)。结论新橙皮苷对破骨细胞的凋亡率以及活力无影响,对其分化具有时间依赖性抑制作用。

Objective To investigate the effect of neohesperidin on apoptosis rate,cell viability and differentiation of osteoclasts. Methods RAW264. 7 cells and mouse bone marrow macrophages were added with receptor activator of nuclear factorκB ligand(RANKL) and neohesperidin respectively,then culture and differentiation were conducted. Apoptosis assay was performed on RAW264. 7 cells. Cell viability assay and time-dependent assay of osteoclasts differentiation were performed on mouse bone marrow macrophages.Results With the increase of neohesperidin concentration,the rate of healthy cells remained above 90% but the necrosis rate and apoptosis rate remained at a low level in each group. There was no statistical difference in the absorptive value between the groups(P > 0. 05). The 0 μM was defined as control group,and the cell viability was not affected by the medicine of each concentration group(P > 0. 05). The average number of osteoclasts in D0 group(NE-),D0 group(NE +),D1 group(NE +),D3 group(NE +),D5 group(NE+) and D7 group(NE +) were 87. 3,82. 7,74,63. 1,61. 7 and 64. 3 respectively. The neohesperidin of10μM added at the 1 st,3 rd,5 th and 7 th day exerted inhibitory effect on the formation of osteoclasts(P < 0. 01),and the most obvious inhibitory effect on the formation of osteoclasts was observed in the neohesperidin added at the 5 th day(P < 0. 001). Conclusion Neohesperidin exerts no effect on the apoptosis rate and viability of the osteoclasts,and obtains a time-dependent inhibition effect on the osteoclasts differentiation,which establishing a foundation for the further research on osteoporosis.