Huang Jing, Wang Yi-Ning, Zhou Yi
State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, Hubei Province, China.
World J Stem Cells. 2023 Aug 26;15(8):807-820. doi: 10.4252/wjsc.v15.i8.807.
Bone marrow mesenchymal stromal cells (BMSCs) are the commonly used seed cells in tissue engineering. Aryl hydrocarbon receptor (AhR) is a transcription factor involved in various cellular processes. However, the function of constitutive AhR in BMSCs remains unclear.
To investigate the role of AhR in the osteogenic and macrophage-modulating potential of mouse BMSCs (mBMSCs) and the underlying mechanism.
Immunochemistry and immunofluorescent staining were used to observe the expression of AhR in mouse bone marrow tissue and mBMSCs. The overexpression or knockdown of AhR was achieved by lentivirus-mediated plasmid. The osteogenic potential was observed by alkaline phosphatase and alizarin red staining. The mRNA and protein levels of osteogenic markers were detected by quantitative polymerase chain reaction (qPCR) and western blot. After coculture with different mBMSCs, the cluster of differentiation (CD) 86 and CD206 expressions levels in RAW 264.7 cells were analyzed by flow cytometry. To explore the underlying molecular mechanism, the interaction of AhR with signal transducer and activator of transcription 3 (STAT3) was observed by co-immunoprecipitation and phosphorylation of STAT3 was detected by western blot.
AhR expressions in mouse bone marrow tissue and isolated mBMSCs were detected. AhR overexpression enhanced the osteogenic potential of mBMSCs while AhR knockdown suppressed it. The ratio of CD86+ RAW 264.7 cells cocultured with AhR-overexpressed mBMSCs was reduced and that of CD206+ cells was increased. AhR directly interacted with STAT3. AhR overexpression increased the phosphorylation of STAT3. After inhibition of STAT3 stattic, the promotive effects of AhR overexpression on the osteogenic differentiation and macrophage-modulating were partially counteracted.
AhR plays a beneficial role in the regenerative potential of mBMSCs partially by increasing phosphorylation of STAT3.
骨髓间充质基质细胞（BMSCs）是组织工程中常用的种子细胞。芳烃受体（AhR）是一种参与多种细胞过程的转录因子。然而，组成型AhR在BMSCs中的功能仍不清楚。
研究AhR在小鼠BMSCs（mBMSCs）成骨和调节巨噬细胞潜能中的作用及其潜在机制。
采用免疫化学和免疫荧光染色观察AhR在小鼠骨髓组织和mBMSCs中的表达。通过慢病毒介导的质粒实现AhR的过表达或敲低。通过碱性磷酸酶和茜素红染色观察成骨潜能。通过定量聚合酶链反应（qPCR）和蛋白质印迹法检测成骨标志物的mRNA和蛋白质水平。与不同的mBMSCs共培养后，通过流式细胞术分析RAW 264.7细胞中分化簇（CD）86和CD206的表达水平。为了探索潜在的分子机制，通过免疫共沉淀观察AhR与信号转导和转录激活因子3（STAT3）的相互作用，并通过蛋白质印迹法检测STAT3的磷酸化。
检测到小鼠骨髓组织和分离的mBMSCs中AhR的表达。AhR过表达增强了mBMSCs的成骨潜能，而AhR敲低则抑制了它。与AhR过表达的mBMSCs共培养的CD86+ RAW 264.7细胞比例降低，CD206+细胞比例增加。AhR直接与STAT3相互作用。AhR过表达增加了STAT3的磷酸化。抑制STAT3后，AhR过表达对成骨分化和巨噬细胞调节的促进作用部分被抵消。
AhR通过增加STAT3的磷酸化在mBMSCs的再生潜能中发挥有益作用。
