TLN1通过激活β-catenin信号通路负性调控骨髓间充质干细胞的成脂分化  

TLN1 negatively regulates adipogenic differentiation of bone marrow mesenchymal stem cells through activating the beta-catenin signaling pathway

作　　者：黄红颜 柯皓腾 叶伟佳 邓华宗 王涛 蔡明希 陈奇凡 岑水忠 Huang Hongyan;Ke Haoteng;Ye Weijia;Deng Huazong;Wang Tao;Cai Mingxi;Chen Qifan;Cen Shuizhong(Department of Breast Surgery,Zhujiang Hospital,Southern Medical University,Guangzhou 510280,Guangdong Province,China;Department of Spinal Surgery,Zhujiang Hospital,Southern Medical University,Guangzhou 510280,Guangdong Province,China)

机构地区：[1]南方医科大学珠江医院乳腺外科,广东省广州市510280 [2]南方医科大学珠江医院脊柱外科,广东省广州市510280

出　　处：《中国组织工程研究》2023年第10期1578-1583,共6页Chinese Journal of Tissue Engineering Research

基　　金：广东省基础与应用基础研究基金自然科学基金面上项目(2022A1515012021),项目负责人:岑水忠;广东省基础与应用基础研究基金青年项目(2020A1515110930),项目负责人:岑水忠;南方医科大学珠江医院院长基金青年培育项目(yzjj2020qn06),项目负责人:岑水忠。

摘　　要：背景:骨髓间充质干细胞成骨-成脂分化平衡在骨代谢疾病中发挥关键作用,课题组前期研究证实TLN1可调控骨髓间充质干细胞成骨分化,但其对骨髓间充质干细胞成脂分化的作用仍未明确。目的:探讨TLN1在骨髓间充质干细胞成脂分化中的作用及机制。方法:诱导骨髓间充质干细胞成脂分化并使用Western blot及qRT-PCR检测TLN1的表达变化;采用CCK-8检测TLN1敲减后骨髓间充质干细胞增殖能力变化,成脂诱导后进行油红O染色及定量分析,qRT-PCR检测骨髓间充质干细胞成脂分化的关键分子PPAR-γ、C/EBP-α的基因表达水平;Western blot检测成脂经典通路蛋白的表达,同时在敲减TLN1的同时添加β-catenin激活剂后检测骨髓间充质干细胞的成脂分化能力,以探讨TLN1调节成脂分化的具体分子机制。结果与结论:TLN1在骨髓间充质干细胞的成脂分化过程中表达下调;敲减TLN1对骨髓间充质干细胞的增殖无明显影响,可显著增强骨髓间充质干细胞成脂诱导分化能力;敲减TLN1可抑制活性的β-catenin表达,而激活β-catenin信号通路可逆转TLN1敲减介导的成脂分化增强。结果表明,TLN1表达在骨髓间充质干细胞成脂分化过程中逐渐下调,敲减TLN1通过抑制β-catenin信号通路发挥促进骨髓间充质干细胞成脂分化的作用。BACKGROUND:The balance between osteogenic and adipogenic differentiation in bone marrow mesenchymal stem cells plays a key role in bone metabolic diseases.Our previous study confirmed that TLN1 can regulate osteogenic differentiation of bone marrow mesenchymal stem cells,but its effect on adipogenic differentiation of bone marrow mesenchymal stem cells remains unclear.OBJECTIVE:To investigate the role and mechanism of TLN1 in the adipogenic differentiation of bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells were induced to adipogenic differentiation and TLN1 expression was detected by q RT-PCR and western blot assay.CCK-8 was used to detect the proliferation of bone marrow mesenchymal stem cells after TLN1 knockdown.The Oil red O staining and quantification were performed after adipogenic induction.Besides,q RT-PCR was used to detect the gene expression levels of the key molecules of adipogenic differentiation of bone marrow mesenchymal stem cells including PPAR-γ and C/EBP-α.Western blot assay was used to detect the classic pathway of adipogenesis.Meanwhile,TLN1 was knockdown and β-catenin activator was added to detect the adipogenic differentiation of bone marrow mesenchymal stem cells,to explore the specific molecular mechanism of TLN1 regulating adipogenesis.RESULTS AND CONCLUSION:TLN1 expression was down-regulated during adipogenic differentiation of bone marrow mesenchymal stem cells.TLN1 knockdown had no significant effect on the proliferation of bone marrow mesenchymal stem cells,but significantly increased the adipogenic induction of bone marrow mesenchymal stem cells.TLN1 knockdown could inhibit the expression of active β-catenin,while activation of β-catenin signaling pathway could reverse the enhancement of adipogenesis mediated by TLN1 knockdown.These findings suggest that TLN1 expression is gradually down-regulated during the adipogenic differentiation of bone marrow mesenchymal stem cells,and TLN1 knockdown can promote the adipogenic differentiation of bone marrow

关 键 词：间充质干细胞 TLN1 成脂分化 Β-CATENIN 信号通路 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R34