摘要
目的探讨二十二碳五烯酸(DPA)对人骨髓间充质干细胞(hBMSCs)向软骨细胞分化的影响。方法通过Binding DB寻找DPA潜在的作用靶点, 利用京都基因及基因组百科全书(KEGG)和Gene ontology(GO)富集DPA靶点调控通路及生物学过程。在hBMSCs诱导成软骨分化中添加DPA, 培养15 d进行实时荧光定量聚合酶链反应(PCR)检测(n=3), 比较DPA组与CON组软骨分化关键基因性别决定区Y框蛋白9(SOX9)和聚蛋白聚糖(ACAN) mRNA的表达量。培养21 d后用鬼笔环肽染色和免疫荧光染色检测细胞骨架和ACAN蛋白分布(n=10), 采用独立样本t检验分析组间差异。结果 DPA具有16个潜在作用靶点, 这些靶点可能参与过氧化物酶体增殖物激活受体信号通路和脂质代谢相关生物学过程。分化中的软骨细胞, SOX9和ACAN的mRNA表达DPA组均高于CON组(1.965±0.118比1.002±0.041, q=9.974, P<0.01)和(1.216±0.018比1.000±0.018, q=3.057, P<0.05)。细胞骨架重建DPA组低于CON组(9.682±0.762比21.450±1.653, t=6.466, P<0.01);而ACAN蛋白积累DPA组高于CON组(12.510±1.215比9.539±0.746, t=2.087, P<0.05)。结论 DPA促进hBMSCs成软骨细胞分化。
Objective To investigate the effect and mechanism of docosapentaenoic acid(DPA)on chondrogenic differentiation of human bone-marrow stem cells(hBMSCs).Methods The potential targets of DPA were identified by Binding DB,and the involved signaling pathways and biological procresses were enriched using Kyoto gene and genome encyclopedia(KEGG)and Gene ontology(GO),respectively.To quantify the impact of DPA on chondrogenic differentiation,the mRNA expression of SRY-box transcription factor 9(Sox9)and aggrecan(ACAN)was detected using real-time polymerase chain reaction(PCR).Phalloidine and immunofluorescence stainings were employed to determine the alterations in the cytoskeletal arrangement and ACAN accumulation by DPA administration.Results A total of 16 DPA targets were identified,which may be involved in the peroxisome proliferator activated receptor(PPAR)signaling pathway and related to lipid metabolism.In the differentiating chondrocytes,the mRNA expression of SOX9 and ACAN was elevated by DPA(1.965±0.118 vs.1.002±0.041,q=9.974,P<0.01)and(1.216±0.018 vs.1.000±0.018,q=3.057,P<0.05).The immunoflourscence of F-actin was weaker in DPA group(9.682±0.762)than in control group(21.450±1.653,t=6.466,P<0.01),and that of ACAN was higher in DPA group(12.510±1.215)than in control group(9.539±0.746,t=2.087,P<0.05).Conclusion DPA can promote chondrogenic differentiation of hBMSCs.
作者
彭海宁
朱东旭
李晓华
魏子然
黄晓红
张英泽
张益
于腾波
Peng Haining;Zhu Dongxu;Li Xiaohua;Wei Ziran;Huang Xiaohong;Zhang Yingze;Zhang Yi;Yu Tengbo(Department of Sports Medicine,the Affiliated Hospital of Qingdao University,Qingdao 266100,China;School of Basic Medicine,Qingdao University,Qingdao 266071,China;Department of Orthopedics,the Third Hospital of Hebei Medical University,Shijiazhuang 050051,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第8期1549-1552,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(31872310、31802022)。
关键词
软骨细胞分化
二十二碳五烯酸
多不饱和脂肪酸
软骨再生
Chondrogenic differentiation
Docosapentaenoic acid
Polyunsaturated fatty acid
Cartilage regeneration
