Department of Orthopedics, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, China.
Cell Cycle. 2021 Oct;20(19):1953-1966. doi: 10.1080/15384101.2021.1969202. Epub 2021 Aug 30.
Neohesperidin (NH) was reported to regulate osteoclastic differentiation, while LncRNA SNHG1 could inhibit osteogenic differentiation of bone marrow stromal cells (BMSCs). In this study, we aimed to explore whether SNHG1-mediated osteogenic differentiation could be regulated by NH. Osteonecrosis and adjacent tissues, as well as normal bone marrow samples were gathered. BMSCs were isolated from normal bone marrow samples by Ficoll density gradient and identified by flow cytometry. Histopathological changes of tissues were detected by hematoxylin-eosin staining. After the treatment with NH or transfection, cell viability, osteogenic differentiation, and the activity of alkaline phosphatase (ALP) in BMSCs were detected by MTT, alizarin red staining, and microplate method, respectively. The histone modification and expressions of SNHG1 and osteogenic marker genes in tissues or BMSCs were detected by q-PCR and Chromatin Immunoprecipitation (ChIp). SNHG1 was highly expressed in osteonecrosis tissues, and typical signs of empty lacunae appeared in the necrotic tissues zone. NH increased viability and osteogenic differentiation of BMSCs, activity of ALP, and expressions of RUNX2, OCN and ALP. NH decreased both SNHG1 expression and H3K4me3 (activating histone modification) occupancies and increased H3K27me3 (inhibiting histone modification) occupancies of SNHG1. Furthermore, siSNHG1 enhanced osteogenic differentiation of BMSCs and expressions of RUNX2, OCN and ALP, while SNHG1 overexpression did the opposite and reversed the effects of NH on the osteogenic differentiation of BMSCs. In a word, NH promotes the osteogenic differentiation of human BMSCs by inhibiting the histone modifications of lncRNA SNHG1.
柚皮苷（NH）被报道能调节破骨细胞分化，而长链非编码 RNA SNHG1 可抑制骨髓基质细胞（BMSCs）的成骨分化。在本研究中，我们旨在探索 NH 是否能调节 SNHG1 介导的成骨分化。收集骨坏死及相邻组织和正常骨髓样本。采用 Ficoll 密度梯度法从正常骨髓样本中分离 BMSCs，并用流式细胞术进行鉴定。苏木精-伊红染色检测组织的组织学变化。用 MTT、茜素红染色和微孔板法分别检测 NH 或转染后 BMSCs 的细胞活力、成骨分化和碱性磷酸酶（ALP）活性。用 q-PCR 和染色质免疫沉淀（ChIp）检测组织或 BMSCs 中的组蛋白修饰和 SNHG1 及成骨标记基因的表达。SNHG1 在骨坏死组织中高表达，坏死组织区出现典型的空陷。NH 增加了 BMSCs 的活力和成骨分化，增加了 ALP 活性和 RUNX2、OCN 和 ALP 的表达。NH 降低了 SNHG1 的表达和 H3K4me3（激活组蛋白修饰）占有率，并增加了 SNHG1 的 H3K27me3（抑制组蛋白修饰）占有率。此外，siSNHG1 增强了 BMSCs 的成骨分化和 RUNX2、OCN 和 ALP 的表达，而 SNHG1 过表达则相反，逆转了 NH 对 BMSCs 成骨分化的影响。总之，NH 通过抑制 lncRNA SNHG1 的组蛋白修饰促进人 BMSCs 的成骨分化。
