乳铁蛋白对三唑酮诱导骨髓间充质干细胞损伤的修复作用

Repairing Effect of Lactoferrin on Triadimefon-Induced Damage of Bone Marrow Mesenchymal Stem Cells

为研究乳铁蛋白(lactoferrin,Lf)对三唑酮所致的骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)损伤的影响,通过构建三唑酮诱导BMSCs损伤模型,采用CCK-8法检测Lf对BMSCs损伤的修复作用;酶联免疫吸附测定法分析成骨过程中碱性磷酸酶(alkaline phosphatase,ALP)、骨钙素(osteocalcin,OCN)的表达情况;同时使用茜素红染色进行成骨分化能力检测,使用油红O染色进行成脂分化能力的检测;反转录实时荧光定量聚合酶链式反应分析成骨和成脂基因表达变化情况。结果表明,与空白对照组相比,不同质量浓度的Lf均可显著促进细胞增殖(P<0.05)。与损伤模型组相比,Lf能提高损伤BMSCs的细胞存活率,增加细胞ALP、OCN质量浓度,增加细胞矿化结节的生成,减少脂滴的生成,上调成骨分化相关基因的相对表达,下调成脂分化相关基因的相对表达。Lf对三唑酮所致损伤的BMSCs有促进增殖作用,能够使BMSCs向成骨方向进行分化,更好地促进新骨形成,从而修复损伤。

The present study was designed to investigate the repairing effect of lactoferrin(Lf)on triadimefon(TDF)-induced damage of bone marrow mesenchymal stem cells(BMSCs).The CCK-8 assay was used to evaluate the proliferation of BMSCs.After being treated with TDF and Lf,the cells were collected for determining the activities of alkaline phosphatase(ALP)and osteocalcin(OCN),and the changes in osteogenic and adipogenic gene expression.The osteogenic differentiation capacity was evaluated using alizarin red staining and adipogenic differentiation using oil red O staining.The expression of adipogenic and osteogenic genes was analyzed by reverse transcription real-time polymerase chain reaction(RT-qPCR).Compared with the blank control group,Lf at all concentrations tested promoted cell proliferation(P<0.05).Lf increased cell survival rate,elevated ALP activity and OCN content in the damaged cells,increased the formation of mineralized nodules,reduced the formation of lipid droplets,increased the relative expression of osteogenic differentiation-related genes,and decreased the relative expression of genes involved in adipogenic differentiation.According to our results,Lf can promote the proliferation of BMSCs damaged by TDF,and the differentiation of bone marrow mesenchymal stem cells into osteogenic direction,thereby promoting the formation of new bone and consequently repairing of cell damage.