益骨汤调控巨噬细胞集落刺激因子信号轴干预骨质疏松大鼠破骨细胞分化的实验研究

Experimental Study on the Efficacy of Yigu Decoction Regulating M-CSF Signal Axis on Osteoclast Differentiation in Osteoporosis Rats

目的:观察益骨汤对骨质疏松大鼠骨组织巨噬细胞集落刺激因子(M-CSF)信号轴的影响,探讨益骨汤通过干预破骨细胞分化治疗骨质疏松症的机制。方法:取12周龄SD雌性大鼠40只,随机分为假手术组、模型组、雌激素组和益骨汤组。药物干预3个月后处死并取双侧股骨,分别进行骨密度测定、生物力学测试、苏木精-伊红(HE)染色观察骨小梁变化,并用Western Blot法和qPCR法检测M-CSF、c-Fms和Grb2表达量的变化。结果:与模型组比较,雌激素组和益骨汤组骨小梁结构较为完整,连续性较好,骨密度均有所升高,差异有统计学意义(P<0.01);弯曲载荷、弹性模量、刚性明显提高,差异有统计学意义(P<0.01)。与假手术组比较,模型组大鼠股骨c-Fms、Grb2、PI3K蛋白的表达增加,差异有统计学意义(P<0.05)。与模型组比较,益骨汤组大鼠股骨组织c-Fms、PI3K蛋白表达明显下降,差异有统计学意义(P<0.01,P<0.05);雌激素组c-Fms蛋白表达明显下降,差异有统计学意义(P<0.01)。假手术组、雌激素组和益骨汤组c-Fms mRNA的表达降低,差异有统计学意义(P<0.01);雌激素组PI3K mRNA的表达降低,差异有统计学意义(P<0.05);益骨汤组Grb2 mRNA的表达降低,差异有统计学意义(P<0.05)。结论:益骨汤可通过调控巨噬细胞集落刺激因子信号轴抑制骨质疏松大鼠破骨细胞分化,减少骨吸收,提高大鼠骨强度。

Objective:To observe the efficacy of Yigu decoction on macrophage colony-stimulating factor(M-CSF)signal axis in bone tissue of osteoporosis rats,and to explore the mechanism of Yigu decoction in treating osteoporosis by intervening osteoclast differentiation.Methods:Forty 12-week-old female SD rats were randomly divided into sham operation group,model group,estrogen group and Yigu decoction group.Rats were killed and bilateral femurs were taken after 3 months of drug intervention.The bone mineral density were measured,the biomechanics were tested,the changes of bone trabeculae were observed by hematoxylin-eosin(HE)staining,and the expressions of M-CSF,c-Fms and Grb2 were detected by Western Blot and qPCR.Results:Compared with the model group,the bone trabecular structure of estrogen group and Yigu decoction group was more complete,the continuity was better,the bone mineral density increased(P<0.01),and the bending load,elastic modulus and rigidity increased significantly(P<0.01).Compared with the sham operation group,the expression of c-Fms,Grb2 and PI3K protein in the femur of the model group increased(P<0.05).Compared with the model group,the expression of c-Fms and PI3K protein in the femur of the Yigu decoction group decreased significantly(P<0.01,P<0.05),and the expression of c-Fms protein in the estrogen group decreased significantly(P<0.01).The expression of c-Fms mRNA decreased in sham operation group,estrogen group and Yigu decoction group(P<0.01),PI3K mRNA decreased in estrogen group(P<0.05),and Grb2 mRNA decreased in Yigu decoction group(P<0.05).Conclusion:Yigu decoction can inhibit osteoclast differentiation,reduce bone resorption and improve bone strength in osteoporosis rats by regulating M-CSF signal axis.