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目的:探讨右归饮对体外培养破骨细胞的分化、凋亡及骨吸收活性的影响。方法:分离培养大鼠破骨细胞,分别以空白对照血清、地塞米松+对照血清、地塞米松+右归饮血清的培养液来培养破骨细胞,通过抗酒石酸酸性磷酸酶(TRAP)染色对TRAP阳性多核破骨细胞计数;采用吖啶橙荧光染色法计数破骨细胞的凋亡率;用RT-PCR测定ATPase a3基因相对表达变化分析破骨细胞骨吸收活性。结果:与空白对照血清组和对照血清+地塞米松组相比,地塞米松+右归饮血清组TRAP染色阳性的破骨细胞数量显著减少(分别为P<0.01,P<0.05),破骨细胞的凋亡率明显增加(P<0.01),ATPase a3基因表达明显减少(P<0.01)。结论:右归饮可抑制破骨细胞的形成和分化及骨吸收活性,促进破骨细胞的凋亡,从而发挥其抗激素性股骨头坏死的作用。
Objective: To investigate the effect of Yougui Yin on differentiation, apoptosis and bone resorption activity of cultured osteoclasts in vitro. Methods: Osteoclasts were isolated from rat osteoclasts. Cultured osteoclasts were cultured with blank control serum, dexamethasone + control serum and dexamethasone + Yugui Yin serum, respectively. The osteoclasts were stained with tartrate - resistant acid phosphatase (TRAP) The number of TRAP-positive multinucleated osteoclasts was counted. Apoptosis rate of osteoclasts was counted by acridine orange fluorescence staining. The relative expression of ATPase a3 gene was detected by RT-PCR. Osteoclast bone resorption activity was analyzed. Results: Compared with control group and dexamethasone group, the number of TRAP-positive osteoclasts in dexamethasone + Yugui Yin serum group was significantly decreased (P <0.01, P <0.05, respectively) The apoptotic rate of osteoblasts was significantly increased (P <0.01), and the expression of ATPase a3 gene was significantly decreased (P <0.01). Conclusion: Yougui Yin can inhibit the formation and differentiation of osteoclasts and bone resorption activity, and promote the apoptosis of osteoclasts, so as to exert its anti-hormonal femoral head necrosis.