Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Department of Orthopaedics, Qilu Hospital of Shandong University, Jinan 250012, China.
Mol Cell Biochem. 2018 Aug;445(1-2):59-65. doi: 10.1007/s11010-017-3251-2. Epub 2017 Dec 15.
For patients with osteoporosis, the inability of osteogenic differentiation is the key reason for bone loss. In this study, we investigated the expression and function of long non-coding RNA BDNF-AS in mesenchymal stem cell-derived osteogenic differentiation. Mouse bone marrow-derived mesenchymal stem cells (BMMSCs) were cultured in vitro and induced toward osteogenic differentiation. Quantitative real-time PCR (qRT-PCR) was used to evaluate gene expressions of BDNF-AS and BDNF during osteogenic differentiation. BMMSCs were also extracted from ovariectomized (OVX) mice. The dynamic change of BDNF-AS in OVX-derived BMMSCs during osteogenic differentiation was also evaluated. Lentivirus was used to upregulate BDNF-AS in BMMSCs. The effects of BDNF-AS upregulation on BMMSCs' proliferation and osteogenic differentiation were then evaluated. In addition, qRT-PCR and western blot were applied to further examine the effect of BDNF-AS upregulation on osteogenesis-associated signaling pathways, including BDNF, OPN, and Runx2, in osteogenic differentiation. BDNF-AS was downregulated, whereas BDNF was upregulated in osteogenic differentiation of BMMSCs. Among OVX-derived BMMSCs, BDNF-AS expression was upregulated during osteogenic differentiation. Lentivirus-induced BDNF-AS upregulation promoted BMMSCs self-proliferation but inhibited osteogenic differentiation, as demonstrated by proliferation, alizarin red staining, and alkaline phosphatase activity assays, respectively. QRT-PCR and western blot demonstrated that BDNF, OPN, and Runx2 were downregulated by BDNF-AS upregulation in the differentiated BMMSCs. BDNF-AS is dynamically regulated in osteogenic differentiation. Upregulating BDNF-AS inhibits osteogenesis, possibly through inverse regulation on BDNF and osteogenic signaling pathways.
对于骨质疏松症患者来说，成骨分化能力不足是骨丢失的关键原因。在本研究中，我们研究了长链非编码 RNA BDNF-AS 在骨髓间充质干细胞源性成骨分化中的表达和功能。体外培养小鼠骨髓间充质干细胞（BMMSCs）并诱导其向成骨分化。采用实时定量 PCR（qRT-PCR）检测BDNF-AS 和 BDNF 在成骨分化过程中的基因表达。还从去卵巢（OVX）小鼠中提取 BMMSCs。还评估了 OVX 来源的 BMMSCs 在成骨分化过程中 BDNF-AS 的动态变化。慢病毒用于上调 BMMSCs 中的 BDNF-AS。然后评估 BDNF-AS 上调对 BMMSCs 增殖和成骨分化的影响。此外，qRT-PCR 和 Western blot 进一步研究了 BDNF-AS 上调对成骨分化相关信号通路（包括 BDNF、OPN 和 Runx2）的影响。在 BMMSCs 的成骨分化过程中，BDNF-AS 下调，而 BDNF 上调。在 OVX 来源的 BMMSCs 中，BDNF-AS 在成骨分化过程中上调。慢病毒诱导的 BDNF-AS 上调促进 BMMSCs 的自我增殖，但抑制成骨分化，分别通过增殖、茜素红染色和碱性磷酸酶活性测定来证明。qRT-PCR 和 Western blot 表明，在分化的 BMMSCs 中，BDNF-AS 上调下调了 BDNF、OPN 和 Runx2。BDNF-AS 在成骨分化中动态调节。上调 BDNF-AS 抑制成骨作用，可能通过对 BDNF 和成骨信号通路的反向调节。
