葛根素对大鼠下颌骨骨髓源巨噬细胞破骨细胞向分化的影响

Effects of puerarin on osteoclast differentiation of marrow-derived macrophages from mandible

目的:分析不同浓度的葛根素对于下颌骨骨髓源巨噬细胞(Bone marrow-derived macrophages,BMMs)破骨分化的影响。方法:体外分离培养4周龄雌性大鼠下颌骨BMMs。通过CCK-8分析1 nmol/L、10 nmol/L、100 nmol/L、1μmol/L、10μmol/L和100μmol/L葛根素对BMMs细胞活性的影响。使用RANKL诱导BMMs破骨分化的同时加入不同浓度葛根素,采用抗酒石酸酸性磷酸酶(TRAP)酶化学染色及q-PCR分析破骨细胞形成及破骨相关基因的改变。采用SPSS16.0软件包对数据进行统计学分析。结果:1 nmol/L、10 nmol/L、100 nmol/L、1μmol/L、10μmol/L葛根素组与对照组活细胞数无显著差异,100μmol/L组活细胞数较对照组降低(P<0.05)。RANKL可诱导BMMs形成多核破骨细胞。10 nmol/L、100 nmol/L及1μmol/L葛根素组破骨细胞数目及破骨细胞面积均较对照组降低(P<0.05)。q-PCR分析发现,10 nmol/L、100 nmol/L及1μmol/L葛根素组NFATc1、C-fos、TRAP及CTSK mRNA表达量均较对照组下降(P<0.05)。结论:葛根素可抑制下颌骨BMMs破骨分化及相关基因表达,为葛根素应用于下颌骨骨质疏松症的预防与治疗提供了依据。

PURPOSE: To study the effect of puerarin on osteoclast differentiation of marrow-derived macrophages(BMMs) from mandible. METHODS: BMMs were obtained from 4-week-old female SD rats. Cell viability of BMMs with different concentration of puerarin was determined by CCK-8. Tartrate resistant acid phosphatase staining and q-PCR were used to analyse the changes of osteoclast formation and expression of osteoclast gene of BMMs due to different concentration of puerarin. Statistical analysis was performed using SPSS 16.0 software package. RESULTS: Cell viability of BMMs with puerarin at 1 nmol/L, 10 nmol/L, 100 nmol/L, 1 μmol/L and 10 μmol/L showed no difference from the control group, and 100 μmol/L group displayed decreased cell viability(P<0.05). Osteoclast number and relative osteoclast surface reduced in BMMs with 10 nmol/L, 100 nmol/L and 1 μmol/L puerarin in comparison with the control group.Furthermore, the expression of NFATc1, C-fos, TRAP and CTSK deceased in BMMs with 10 nmol/L, 100 nmol/L,and 1 μmol/L puerarin. CONCLUSIONS: Puerarin could impede osteoclast differentiation of mandibular BMMs, which may be used to prevent and treat osteoporosis in alveolar and jaw bone.