[1]周年,林鑫,陆杨.SIRT1通过β-catenin信号调控干细胞成软骨与成骨分化[J].第三军医大学学报,2020,42(13):1308-1314.
　HOU Nian,LIN Xin,LU Yang.SIRT1 regulates osteogenic and chondrogenic differentiation of stem cells by β-catenin signal[J].J Third Mil Med Univ,2020,42(13):1308-1314.
点击复制

SIRT1通过β-catenin信号调控干细胞成软骨与成骨分化(/HTML )
分享到：

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第13期
页码:
1308-1314
栏目:
基础医学
出版日期:
2020-07-15

文章信息/Info

Title:
SIRT1 regulates osteogenic and chondrogenic differentiation of stem cells by β-catenin signal
作者:
周年; 林鑫; 陆杨
重庆医科大学附属第一医院骨科
Author(s):
HOU Nian;  LIN Xin;  LU Yang

Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
干细胞; 成骨分化; 成软骨分化
Keywords:
stem cells;  chondrogenic differentiation;  osteogenic differentiation
分类号:
R322.71; R329.21; R345
文献标志码:
A
摘要:

目的探讨SIRT1对干细胞成软骨、成骨分化的调控作用及机制。方法C3H10T1/2感染腺病毒Ad-SIRT1后于普通培养基培养，通过荧光显微镜了解腺病感染情况，采用Western blot验证目的蛋白是否高表达；在不同时间点，通过Western blot、阿尔新蓝染色、碱性磷酸酶染色检测干细胞成骨、成软骨分化关键转录调控因子及相关分化标志物的表达情况；使用XAV-939抑制β-catenin信号后，再利用Western blot检测成骨、成软骨分化标志物表达情况。结果C3H10T1/2感染腺病毒Ad-SIRT1后，SIRT1获得高表达；在诱导分化后第5、7、9天，Ad-SIRT1组成软骨分化的标志物Ⅱ型胶原、蛋白多糖，以及成骨分化的标志物Ⅰ型胶原、骨钙素较Ad-GFP组明显增加（P<0.05）；阿尔新蓝染色结果发现Ad-SIRT1组软骨细胞外基质分泌较Ad-GFP组明显增加；碱性磷酸酶染色结果发现Ad-SIRT1组部分细胞着色为阳性，而Ad-GFP组无明显着色的细胞；同时发现诱导分化后的第3、5天，Ad-SIRT1组中成软骨、成骨分化关键转录调控因子Sox9、Runx2的表达较Ad-GFP组明显增加（P<0.05）；Ad-SIRT1组与Ad-GFP组比较，β-catenin乙酰化水平明显降低(P<0.05)，而抑制β-catenin信号后，与Ad-SIRT1组比较，Ad-SIRT1+XAV-939组干细胞成软骨及成骨分化标志物表达明显降低（P<0.05）。结论SIRT1能够通过β-catenin信号促进干细胞的成软骨及成骨分化。

Abstract:

ObjectiveTo explore the regulative role of SIRT1 in chondrogenic and osteogenic differentiation of stem cells and investigate its underlying mechanism. MethodsPlasmid Ad-SIRT1 was used to infect C3H10T1/2 cells to express SIRT1 exogenously, and its expression were detected by fluorescence microscopy and Western blotting. The chondrogenic and osteogenic differentiation key transcription regulators and differentiation markers were detected by Western blotting, Alcian Blue staining and Alkaline Phosphatase staining at different time points. The differentiation markers were also detected by Western blotting after β-catenin inhibited by XAV-939. ResultsSIRT1 was overexpressed in C3H10T1/2 cells after Ad-SIRT1 infection. The Ad-SIRT1 infected cells exhibited significantly elevated protein levels of the chondrogenic (type Ⅱ collagen and aggrecan) and osteogenic markers (type Ⅰ collagen and osteocalcin) when compared to Ad-GFP group in 5, 7 and 9 d after differentiation induction (P<0.05). The C3H10T1/2 cells after Ad-SIRT1 infection showed enhanced secretion of cartilaginous matrix by Alcian Blue staining, and more cells were positive to alkaline phosphatase staining, but no such stained cells were observed in the cells infected with Ad-GFP. In 3 and 5 d after differentiation, the chondrogenic and osteogenic key transcription regulators Sox9 and Runx2 were significantly increased at the protein level in Ad-SIRT1 infected cells than the Ad-GFP infected cells (P<0.05). The level of acetylated β-catenin was also significantly decreased (P<0.05), and XAV-939 treatment decreased above chondrogenic and osteogenic differentiation markers (P<0.05). ConclusionSIRT1 can promote chondrogenic and osteogenic differentiation by β-catenin signal.

相似文献/References:

[1]朱灏,许建中,周强,等.人骨髓间充质干细胞体外增殖及成骨分化的增龄变化[J].第三军医大学学报,2005,27(01):59.
[2]王志平,柳峰,赵建华.脉冲电磁场对脱钙骨基质诱导人骨髓干细胞成骨分化的影响[J].第三军医大学学报,2010,32(09):947.
　Wang Zhiping,Liu Feng,Zhao Jianhua.Effects of pulsed electromagnetic fields on osteogenic differentiation in demineralized bone matrix-induced human marrow stromal cells in vitro[J].J Third Mil Med Univ,2010,32(13):947.
[3]伍秋香,任春美,陈振华,等.PTEN在骨形态发生蛋白9诱导小鼠胚胎成纤细胞骨向分化中的作用[J].第三军医大学学报,2016,38(03):264.
　Wu Qiuxiang,Ren Chunmei,Chen Zhenhua,et al.Role of PTEN in bone morphogenetic protein 9 induced osteogenic differentiation in mouse embryonic fibroblasts[J].J Third Mil Med Univ,2016,38(13):264.
[4]蒋琳,周鹏飞,王佳,等.BMPs-ERK5信号通路调控人牙周膜干细胞成骨分化的研究[J].第三军医大学学报,2016,38(07):718.
　Jiang Lin,Zhou Pengfei,Wang Jia,et al.Bone morphogenetic proteins-ERK5 signaling pathway mediates osteogenic differentiation of human periodontal ligament stem cells[J].J Third Mil Med Univ,2016,38(13):718.
[5]常慧君,申涛,董世武,等.purmorphamine促进人牙周膜干细胞应力成骨[J].第三军医大学学报,2012,34(09):839.
　Chang Huijun,Shen Tao,Dong Shiwu,et al.Purmorphamine promotes osteogenic differentiation in human periodontal ligament stem cells under dynamic tensile[J].J Third Mil Med Univ,2012,34(13):839.
[6]向强,张瑗,崔翔,等.一种具有双嗜性功能融合蛋白FN/CDH的制备及其促粘附、成骨生物活性鉴定[J].第三军医大学学报,2013,35(23):2536.
　Xiang Qiang,Zhang Yuan,Cui Xiang,et al.Preparation of FN/CDH amphotropic recombinant protein and its capacity in promoting osteoblastic adhesion and ossification[J].J Third Mil Med Univ,2013,35(13):2536.
[7]李锐冬,邓忠良,张亚梅,等.不同剂量TGF-β1在BMP9诱导间充质干细胞成骨分化中的作用[J].第三军医大学学报,2014,36(02):98.
　Li Ruidong,Deng Zhongliang,Zhang Yamei,et al.Effects of different concentrations of TGF-β1 on BMP9-induced osteogenesis in mesenchymal stem cells[J].J Third Mil Med Univ,2014,36(13):98.
[8]黄科,胡宗凯,喻胜鹏,等.Cbfβ人工miRNA干扰载体构建及稳定细胞株系的获得与鉴定[J].第三军医大学学报,2014,36(07):682.
　Huang Ke,Hu Zongkai,Yu Shengpeng,et al.Construction of artificial microRNA vector interfering mouse Cbfβ gene and establishment and identification of stable cell line[J].J Third Mil Med Univ,2014,36(13):682.
[9]周年,黄伟,廖军义,等.低氧诱导因子-1α对骨形态发生蛋白2诱导的干细胞成软骨、成骨分化的影响[J].第三军医大学学报,2014,36(12):1243.
　Zhou Nian,Huang Wei,Liao Junyi,et al.HIF-1α potentiates BMP2-induced chondrogenic differentiation but inhibits osteogenic differentiation in stem cells[J].J Third Mil Med Univ,2014,36(13):1243.
[10]郭志良,滕海军,王亮,等.恒磁场对培养的骨髓间充质干细胞成骨分化的影响及机制[J].第三军医大学学报,2014,36(12):1255.
　Guo Zhiliang,Teng Haijun,Wang Liang,et al.Effects of static magnetic field on osteogenesis of bone marrow mesenchymal stem cells[J].J Third Mil Med Univ,2014,36(13):1255.

更新日期/Last Update: 2020-07-06