姜黄素调控HO-1改善高糖环境下骨髓间充质干细胞的成骨分化  

Curcumin promotes osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose environment by regulating HO-1

作　　者：魏咸亭 陈宝康 董鑫 闫康 张小平 廖博 WEI Xian-Ting;CHEN Bao-Kang;DONG Xin;YAN Kang;ZHANG Xiao-ping;LIAO Bo(School of Basic Medical Sciences,Air Force Medical University,Xi´an haanxi 710032,China;Department of Orthopaedics,the Second Affiliated Hospital of Air Force Medical University,Xi´an Shaanxi 710038,China)

机构地区：[1]空军军医大学基础医学院,陕西西安710032 [2]空军军医大学第二附属医院骨科,陕西西安710038

出　　处：《局解手术学杂志》2024年第9期783-787,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：国家自然科学基金(82174166);陕西省自然科学基础研究计划项目(2023-JC-JQ-70)。

摘　　要：目的探讨姜黄素对高糖环境下人骨髓间充质干细胞(hBMSCs)成骨分化的影响及其机制。方法将培养的hBMSCs分为正常组、高糖组和高糖+姜黄素组。通过检测碱性磷酸酶(ALP)活性评估各组细胞早期成骨分化水平;茜素红染色评估成骨分化晚期矿化结节的形成情况;RT-PCR检测成骨诱导分化21 d后成骨相关基因Runt相关转录因子2(Runx2)、骨钙素(OCN)和Ⅰ型胶原蛋白(COL-1)的表达;采用Western blot检测各组细胞中血红素氧合酶-1(HO-1)的表达;进一步构建HO-1小干扰RNA(siRNA)模型并检测干扰效率,对比高糖+姜黄素组和高糖+姜黄素+siHO-1组成骨相关蛋白(Runx2、OCN和COL-1)的表达水平。结果与正常组比较,高糖组细胞ALP活性降低,矿化结节形成减少,成骨相关基因(Runx2、OCN、COL-1)表达下降,HO-1的表达受到抑制(P<0.05);与空载体组相比,siHO-1组HO-1表达显著下降,表明siRNA干扰成功(P<0.01)。相比高糖+姜黄素组,高糖+姜黄素+siHO-1组成骨相关蛋白(OCN、COL-1和Runx2)的表达水平均下降(P<0.05)。结论姜黄素可改善高糖环境下hBMSCs的成骨分化能力,且与HO-1的表达有关。Objective To study the effect of curcumin on osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs)in high glucose condition and its mechanism.Methods The cultured hBMSCs were divided into the normal group,high glucose group,and high glucose+curcumin group.The early osteogenic differentiation level of the cells in each group was assessed by detecting alkaline phosphatase(ALP)activity.Alizarin red staining was used to evaluate the formation of mineralized nodules in the late stage of osteo‐genic differentiation.The expression of osteogenic-related genes,including Runt-related transcription factor 2(Runx2),osteocalcin(OCN),and typeⅠcollagen(COL-1),was detected by RT-PCR after 21 days of osteogenic induction.Western blot was used to detect the expression of heme oxygenase-1(HO-1)in each group.Furthermore,an HO-1 small interfering RNA(siRNA)model was constructed and its interference efficiency was assessed.The expression levels of osteogenesis-related proteins(Runx2,OCN,and COL-1)between the high glucose+curcumin group and high glucose+curcumin+siHO-1 group were compared.Results Compared with the normal group,the high glucose group showed decreased ALP activity,reduced formation of mineralized nodules,decreased expression of osteogenic-related genes(Runx2,OCN,and COL-1),and inhibited expression of HO-1(P<0.05).Compared with the empty vector group,the siHO-1 group showed significantly reduced expression of HO-1 in cells,indicating successful siRNA interference(P<0.01).Compared with the high glucose+curcumin group,the expression levels of osteogenesis-related proteins(OCN,COL-1,and Runx2)were all decreased in the high glucose+curcumin+siHO-1 group(P<0.05).Conclusion Curcumin can promote osteogenic differentiation of hBMSCs under high glucose environment,which is related to the expression of HO-1.

关 键 词：姜黄素 人骨髓间充质干细胞 成骨分化 血红素氧合酶-1 

分 类 号：R285.5[医药卫生—中药学] R329.2[医药卫生—中医学]