木犀草素对小鼠体外培养前成骨细胞MC3T3-E1增殖、分化、矿化和Wnt通路的影响

Effect of luteolin on the proliferation,differentiation,mineralization and Wnt pathway of MC3T3-E1 osteoblasts in vitro

目的:探讨木犀草素对小鼠体外培养前成骨细胞MC3T3-E1增殖、分化和矿化及Wnt通路的影响。方法:在体外培养的MC3T3-E1s培养基中加入木犀草素0.25~8.00μmol/L培养12、24和48 h,以洛伐他汀0.04μmol/L作为阳性对照,采用细胞计数试剂盒法检测细胞增殖情况;用碱性磷酸酶(ALP)试剂盒检测细胞内ALP的活性;茜素红S染色法检测细胞矿化能力;采用实时荧光定量反转录PCR法检测细胞Ⅰ型胶原、骨钙素、低密度脂蛋白受体相关蛋白5(Lrp5)、β-连环素、护骨素/细胞核因子κB受体活化因子配体mRNA表达水平。结果:木犀草素可以剂量依赖性和时间依赖性地促进成骨细胞MC3T3-E1增殖(P<0.05~P<0.01);木犀草素(2.00、4.00和8.00μmol/L)可以剂量依赖性地促进MC3T3-E1成骨细胞ALP活性,增强MC3T3-E1成骨细胞矿化能力并上调MC3T3-E1成骨细胞中Ⅰ型胶原、骨钙素、Lrp5、β-连环素和护骨素/细胞核因子κB受体活化因子配体mRNA表达。结论:木犀草素2.00、4.00和8.00μmol/L可以促进MC3T3-E1细胞的增殖、分化与矿化及Wnt通路中Lrp5和β-连环素mRNA的表达。

Objective： To investigate the effects of luteolin on the proliferation, differentiation, mineralization and Wnt pathway of MC3T3-E1 osteoblasts in vitro. Methods： The MC3T3-E1 osteoblasts were cultured with the 0.25 to 8.00 μ mol/L of luteolin for 12,24 and 48 h in vitro. The 0.04 txmol/L of lovastatin treating cell was used as the positive control group. The proliferation, activity of alkaline phosphatase（ALP） and mineralization capacity of MC3T3-E1 osteoblasts in vitro were detected by cell counting kits-8 method, alkaline phosphatase kit and Alizarin Red S staining,respectively. The mRNA expressions of type I collagen, osteocalcin,low density lipoprotein receptor related protein 5 （ Lrp5 ）, β-catenin, osteoprotegerin and nuclear factor KB-activating factor receptor ligands were quantified by real-time reverse transcription PCR （qPCR）. Results： Luteolin could improve the proliferation of MC3T3-E1 osteoblasts with time- and dose-dependence（ P 〈 0.05 to P 〈 0.01 ）. Lutenlin could promote the activity of ALP, strengthen the mineralization capacity and upregulate the mRNA expressions of type I collagen,osteocalcin, Lrp5, β-catenin, osteoprotegerin and nuclear factor κB- activating factor receptor ligands in MC3T3-Els with dose-dependence in vitro. Conclusions： The 2.00,4.00 and 8.00 μmol/L of luteolin can promote the proliferation, differentiation and mineralization and the expressions pf Lrp5 and β-catenin in MC3T3-E1 osteoblasts.