Function and regulation of transforming growth factor β1 signalling in antler chondrocyte proliferation and differentiation
Function and regulation of transforming growth factor β1 signalling in antler chondrocyte proliferation and differentiation
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DOI：
10.1111/cpr.12637
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发表时间：
2019-07-01
影响因子：
8.5
通讯作者：
Yue, Zhan-Peng
中科院分区：
文献类型：
--
作者：
Ma, Li;Yang, Zhan-Qing;Yue, Zhan-Peng
目的软骨细胞的增殖和分化是软骨内成骨的关键，但其调控机制尚不清楚。本研究旨在探讨TGF β 1信号通路在鹿茸软骨细胞增殖分化中的生理功能及其与Notch、Shh信号通路和Foxa的关系。材料与方法采用免疫荧光、Western blot、MTS试验、流式细胞术、RNA干扰和实时荧光定量PCR等方法，分析TGF β 1信号通路在鹿茸软骨细胞增殖和分化中的作用和调控机制。结果TGF β 1、TGF β R 1和TGF β R 2在鹿茸软骨中呈高表达。TGF β 1促进软骨细胞增殖，增加S期细胞的比例，并诱导肥大软骨细胞标志物Col X、Runx 2和Alpl的表达。然而，TGF β受体抑制剂SB 431542和Smad 3抑制剂SIS 3减弱了这种诱导作用。同时，TGF β 1激活Notch和Shh信号传导，其阻断减弱了rTGF β 1的上述作用，而rShh的加入挽救了由SB 431542和SIS 3引起的软骨细胞增殖和分化的缺陷。进一步的分析显示，Notch信号传导的抑制阻碍了TGF β 1对Shh通路的激活。Foxa 1、Foxa 2和Foxa 3的敲除消除了TGF β 1对软骨细胞分化的作用。Notch和Shh信号传导介导TGF β 1对Foxa转录因子的调节。结论TGF β 1信号通过Notch-Shh-Foxa途径诱导鹿茸软骨细胞增殖分化。
Objectives Chondrocyte proliferation and differentiation are crucial for endochondral ossification, but their regulatory mechanism remains unclear. The present study aimed to determine the physiological function of TGF beta 1 signalling in the proliferation and differentiation of antler chondrocytes and explore its relationship with Notch, Shh signalling and Foxa. Materials and methods Immunofluorescence, Western blot, MTS assay, flow cytometry, RNA interference and real-time PCR were used to analyse the function and regulatory mechanisms of TGF beta 1 signalling in antler chondrocyte proliferation and differentiation. Results TGF beta 1, TGFBR1 and TGFBR2 were highly expressed in antler cartilage. TGF beta 1 promoted chondrocyte proliferation, increased the proportion of S-phase cells and induced the expression of hypertrophic chondrocyte markers Col X, Runx2 and Alpl. However, this induction was weakened by TGF beta receptor inhibitor SB431542 and Smad3 inhibitor SIS3. Simultaneously, TGF beta 1 activated Notch and Shh signalling whose blockage attenuated the above effects of rTGF beta 1, whereas addition of rShh rescued the defects in chondrocyte proliferation and differentiation elicited by SB431542 and SIS3. Further analysis revealed that inhibition of Notch signalling impeded TGF beta 1 activation of the Shh pathway. Knockdown of Foxa1, Foxa2 and Foxa3 abrogated the effects of TGF beta 1 on chondrocyte differentiation. Notch and Shh signalling mediated the regulation of Foxa transcription factors by TGF beta 1. Conclusions TGF beta 1 signalling could induce the proliferation and differentiation of antler chondrocytes through Notch-Shh-Foxa pathway.