Meng Xiangsheng, Zhu Haiquan
Trauma Center, Lianyungang City No.1 People's Hospital, Lianyungang, China.
J Invest Surg. 2023 Dec;36(1):2197054. doi: 10.1080/08941939.2023.2197054.
In this study, we aimed to explore whether the SRY-box transcription factor 9 (SOX9) can play protective roles against the occurrence and development of osteonecrosis of the femoral head (ONFH) by regulating the proliferation, apoptosis, and osteogenic differentiation of human bone marrow stromal cells (hBMSCs) via the Wnt/β-catenin pathway. We used 1600 mg of glucocorticoid (GC) to induce hBMSCs to establish an ONFH cell model and performed various experiments. Reverse transcription-quantitative polymerase chain reaction and western blotting assays were used to determine the expression levels of SOX9 and osteoblast markers, such as the RUNX family transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osterix, Wnt3a, and β-catenin. An ALP detection kit was used to measure the ALP activity. Flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were performed to determine the cell viability. GC treatment decreased the expression levels of RUNX2, ALP, and osterix, suppressed ALP activity, and inhibited SOX9 expression. SOX9 overexpression promoted GC-induced cell proliferation and decreased cell apoptosis. Additionally, hBMSCs were transfected with SOX9-small interfering RNA in GC treatment, and SOX9 knockdown was found to suppress the osteogenic differentiation of cells and decrease their viability. Our results revealed that SOX9 is related to the Wnt/β-catenin pathway in ONFH. Moreover, SOX9 participated in ONFH development by activating the Wnt/β-catenin pathway.
在本研究中，我们旨在探讨SRY盒转录因子9（SOX9）是否能通过Wnt/β-连环蛋白途径调节人骨髓间充质干细胞（hBMSCs）的增殖、凋亡和成骨分化，从而对股骨头坏死（ONFH）的发生和发展起到保护作用。我们使用1600mg糖皮质激素（GC）诱导hBMSCs建立ONFH细胞模型并进行了各种实验。采用逆转录-定量聚合酶链反应和蛋白质印迹分析来测定SOX9和成骨标志物的表达水平，如RUNX家族转录因子2（RUNX2）、碱性磷酸酶（ALP）、osterix、Wnt3a和β-连环蛋白。使用ALP检测试剂盒测量ALP活性。通过流式细胞术和3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐分析来测定细胞活力。GC处理降低了RUNX2、ALP和osterix的表达水平，抑制了ALP活性，并抑制了SOX9的表达。SOX9过表达促进了GC诱导的细胞增殖并减少了细胞凋亡。此外，在GC处理中用SOX9小干扰RNA转染hBMSCs，发现敲低SOX9可抑制细胞的成骨分化并降低其活力。我们的结果表明，SOX9与ONFH中的Wnt/β-连环蛋白途径相关。此外，SOX9通过激活Wnt/β-连环蛋白途径参与ONFH的发展。
