姜黄素促进炎症微环境下大鼠骨髓间充质干细胞骨向分化

Curcumin promotes bone differentiation of rat bone marrow mesenchymal stem cells in inflammatory microenvironment

目的研究姜黄素对炎症微环境下大鼠骨髓间充质干细胞(BMSCs)成骨分化的影响。方法分离、培养大鼠BMSCs,以10 ng/mL的肿瘤坏死因子α(TNF-α)诱发细胞炎症反应,用不同浓度姜黄素处理细胞,CCK-8检测细胞增殖以便筛选出实验所需姜黄素适宜浓度;实验分组为正常组、炎症组和炎症+姜黄素组;碱性磷酸酶(ALP)和茜素红染色检测成骨分化,实时荧光定量PCR检测骨涎蛋白(BSP)、Runt相关转录因子2(Runx2)的表达。结果成功分离培养BMSCs;低浓度的姜黄素无细胞毒性,1μmol/L为适宜浓度;炎症降低大鼠BMSCs成骨相关基因BSP、Runx2表达,且ALP染色变浅、矿化结节减少;而姜黄素则可以逆转这种趋势,促进炎症状态下成骨相关基因BSP、Runx2的表达(P<0.05),较炎症组ALP染色加深,矿化结节增多。结论姜黄素可以促进炎症微环境下BMSCs成骨分化。

Objective To study the effect of curcumin on the osteogenic differentiation of rat bone marrow mesenchymal stem cells(BMSCs)under inflammatory microenvironment.Methods BMSCs were isolated and cultured,10ng/ml TNF-αwas used to induce cell inflammation,cells were treated with different concentrations of curcumin,and cell proliferation was detected by CCK-8 to select the appropriate concentration of curcumin for the experiment;the cells were divided into normal group,inflammation group and curcumin+inflammatory group;osteogenic differentiation was dectected with alkaline phosphatase(ALP)and alizarin red staining;the expression of bone sialoprotein(BSP)and runt-related transcription factor 2(Runx2)was detected by real-time fluorescent quantitative PCR.Result Bone marrow mesenchymal stem cells were successfully isolated and cultured;the low concentration of curcumin had no cytotoxicity,and 1μmol/L was the optimum concentration.Inflammation decreased the expression of BSP and runx2,ALP staining and mineralized nodules were reduced;however,curcumin can reverse this trend and increase the expression of BSP and runx2 in the inflammatory state(P<0.05),and the ALP staining was deeper and the mineralized nodules increased compared with the inflammatory group.Conclusion Curcumin can promote osteogenic differentiation of bone marrow mesenchymal stem cells under inflammatory microenvironment.