小分子药物TD-198946促进大鼠骨髓间充质干细胞的成骨分化  

Small molecule drug TD-198946 enhances osteogenic differentiation of rat bone marrow mesenchymal stem cells

作　　者：杨超[1] 罗宗平[1] Yang Chao;Luo Zongping(Department of Orthopedics,First Affiliated Hospital of Soochow University,Institute of Orthopedics of Soochow University,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院骨科,苏州大学骨科研究所,江苏省苏州市215006

出　　处：《中国组织工程研究》2025年第13期2648-2654,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(32071307),项目负责人:罗宗平。

摘　　要：背景:小分子药物TD-198946是一种能诱导干细胞形成软骨的高效软骨生成剂,但目前尚不清楚其对成骨分化的作用。目的:探讨小分子药物TD-198946促进大鼠骨髓间充质干细胞成骨分化的效果及其作用机制。方法:提取SD大鼠骨髓间充质干细胞,利用CCK-8法评估不同浓度TD-198946对骨髓间充质干细胞增殖的影响,确定TD-198946的最佳使用浓度;然后加入最适浓度TD-198946和成骨分化培养基对骨髓间充质干细胞进行成骨诱导;成骨诱导第3天用qRT-PCR检测碱性磷酸酶、Runt相关转录因子2、骨桥蛋白、骨钙蛋白、Ⅰ型胶原基因表达;成骨诱导第7天进行碱性磷酸酶染色,Western blot检测Runt相关转录因子2、Ⅰ型胶原、AKT、p-AKT、PI3K、p-PI3K蛋白表达;成骨诱导第21天进行茜素红染色。结果与结论:①CCK-8实验结果显示,100 nmol/L TD-198946能促进骨髓间充质干细胞增殖;②碱性磷酸酶染色及茜素红染色结果显示,TD-198946能促进大鼠骨髓间充质干细胞的成骨分化;③qRT-PCR结果显示,TD-198946可促进成骨相关基因碱性磷酸酶、Runt相关转录因子2、骨桥蛋白、骨钙蛋白、Ⅰ型胶原的表达;④Western blot结果显示,TD-198946可促进Runt相关转录因子2、Ⅰ型胶原以及p-PI3K、p-AKT蛋白表达。结果表明,小分子药物TD-198946可能通过激活PI3K/AKT信号通路,诱导大鼠骨髓间充质干细胞成骨分化。BACKGROUND:TD-198946 is a small molecule drug known to induce stem cells to form cartilage;however,its effect on osteogenic differentiation remains unclear.OBJECTIVE:To investigate the effects of the small molecule drug TD-198946 on promoting osteoblastic differentiation of rat bone marrow mesenchymal stem cells and its mechanism of action.METHODS:Bone marrow mesenchymal stem cells(BMSCs)were extracted from SD rats.CCK-8 assay was used to evaluate the effect of different concentrations of TD-198946 on the proliferation of bone marrow mesenchymal stem cells to determine the optimal concentration of TD-198946.Then,the optimal concentration of TD-198946 and osteogenic medium were added to induce osteogenic differentiation of bone marrow mesenchymal stem cells.On day 3,qRTPCR was used to detect the expression of alkaline phosphatase,Runt-related transcription factor 2,osteopontin,osteocalcin,and type I collagen genes.On day 7 of osteogenic induction,alkaline phosphatase staining and western blot assay were performed to detect the expression of Runt-related transcription factor 2,type I collagen,AKT,p-AKT,PI3K,and p-PI3K proteins.On day 21 of osteogenic induction,Alizarin red staining was conducted.RESULTS AND CONCLUSION:(1)CCK-8 assay results revealed that 100 nmol/L TD-198946 could promote the proliferation of bone marrow mesenchymal stem cells.(2)The results of alkaline phosphatase staining and alizarin red staining showed that TD-198946 could promote osteogenic differentiation of rat bone marrow mesenchymal stem cells.(3)qRT-PCR results showed that TD-198946 could promote the expression of osteogenic genes alkaline phosphatase,Runt-related transcription factor 2,osteopontin,osteocalcin,and type I collagen.(4)Western blot assay results showed that TD-198946 could enhance the expression of Runt-related transcription factor 2,type I collagen,p-PI3K,and p-AKT.The results indicate that the small molecule drug TD-198946 may induce osteogenic differentiation of rat bone marrow mesenchymal stem cells by activating the PI3

关 键 词：骨髓间充质干细胞 TD-198946 成骨分化 PI3K/AKT信号通路 AKT P-AKT 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学] R318