Kartogenin对大鼠和兔源骨髓间充质干细胞成软骨与成骨分化的差异性影响  

Differential effects of kartogenin on chondrogenic and osteogenic differentiation of rat and rabbit bone marrow mesenchymal stem cells

作　　者：柳承源 过倩萍[1,2] Liu Chengyuan;Guo Qianping(Department of Orthopedic Surgery,First Affiliated Hospital,Soochow University,Suzhou 215000,Jiangsu Province,China;Orthopedic Institute,Suzhou Medical College,Soochow University,Suzhou 215000,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院骨科,江苏省苏州市215000 [2]苏州大学苏州医学院骨科研究所,江苏省苏州市215000

出　　处：《中国组织工程研究》2025年第35期7490-7498,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(82072424),项目负责人:过倩萍。

摘　　要：背景:骨髓间充质干细胞具有多向分化潜力,是软骨和骨组织再生研究中的重要细胞来源。Kartogenin(KGN)是一种小分子药物,已被证明能够促进干细胞成软骨分化,但促成骨分化能力仍存在争议,而且KGN对不同种属干细胞成软骨、成骨分化的具体作用仍未得到充分阐述。目的:探讨KGN对兔源和大鼠源骨髓间充质干细胞成软骨和成骨分化的影响。方法:通过全骨髓分离贴壁法获得兔源和大鼠源骨髓间充质干细胞,分别用不同浓度KGN进行处理,采用CCK-8法检测细胞增殖情况,采用甲苯胺蓝和茜素红染色评估成软骨和成骨分化情况。筛选出合适浓度的KGN后,使用碱性磷酸酶染色、qRT-PCR分析和Western blot分析成骨、成软骨相关基因和蛋白的表达。结果与结论:①在0-10000 nmol/L范围内,KGN对兔源和大鼠源骨髓间充质干细胞的增殖没有显著影响;②KGN对兔源和大鼠源骨髓间充质干细胞的分化效果却表现出显著差异,在兔源骨髓间充质干细胞中,KGN主要促进成软骨分化,同时抑制成骨分化,具体表现为甲苯胺蓝染色呈阳性,而碱性磷酸酶染色和茜素红染色没有明显变化,qRT-PCR分析显示KGN上调了成软骨相关基因(如Col2a1和Sox9)的表达,并下调了成骨相关基因(如Alpl、Col1a1、Runx2和Bglap)的表达,Western blot实验也有相似的结果;③相比之下,在大鼠源骨髓间充质干细胞中,KGN同时促进了成软骨和成骨分化,尽管甲苯胺蓝染色变化不明显,但碱性磷酸酶染色和茜素红染色阳性率显著增加,表明成骨分化增强,qRT-PCR分析显示KGN上调了成软骨相关基因(如Col2a1和Sox9)的表达,同时上调了成骨相关基因(如Alpl、Col1a1、Runx2和Bglap)的表达,Western blot实验也有相似的结果。这些结果表明,KGN在兔源和大鼠源骨髓间充质干细胞中的分化调控作用存在明显差异,可能与两种细胞中的基因表达模式和信号通路不同有关。BACKGROUND:Bone marrow mesenchymal stem cells possess multipotent differentiation potential and are an important source of cells for cartilage and bone tissue regeneration research.Kartogenin is a small-molecule drug that has been demonstrated to promote stem cell differentiation towards chondrogenesis.However,the ability to promote osteogenic differentiation is still controversial,and the specific effects of kartogenin on the chondrogenic and osteogenic differentiation of stem cells of different species have not been fully elucidated.OBJECTIVE:To investigate the effects of kartogenin on chondrogenic and osteogenic differentiation in rabbit-derived and rat-derived bone marrow mesenchymal stem cells.METHODS:Rabbit-derived and rat-derived bone marrow mesenchymal stem cells were obtained through whole bone marrow separation and adherence methods,and were treated with varying concentrations of kartogenin.Cell proliferation was detected using the CCK-8 assay.Chondrogenic and osteogenic differentiation was assessed via toluidine blue and alizarin red staining,respectively.After screening for the optimal concentration of kartogenin,alkaline phosphatase staining,qRT-PCR,and western blot assay were employed to analyze the expression of osteogenic and chondrogenic-related genes and proteins.RESULTS AND CONCLUSION:(1)Within the concentration range of 0 to 10000 nmol/L,kartogenin did not significantly affect the proliferation of rabbit-derived or mouse-derived bone marrow mesenchymal stem cells.(2)The differentiation effect of kartogenin on rabbit-derived and rat-derived bone marrow mesenchymal stem cells showed significant differences.In rabbit-derived bone marrow mesenchymal stem cells,kartogenin predominantly promoted chondrogenic differentiation while inhibiting osteogenic differentiation.This was evident from positive toluidine blue staining,whereas alkaline phosphatase and alizarin red staining exhibited no significant changes.qRT-PCR analysis revealed upregulation of chondrogenic-related genes(Col2a1 and Sox9)and down

关 键 词：骨髓间充质干细胞 Kartogenin 成软骨分化 成骨分化 基因表达 细胞增殖 再生医学 工程化干细胞 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R683