小核仁RNA宿主基因5对骨髓间充质干细胞成骨分化和凋亡的影响

Small nucleolar RNA host gene 5 involved in regulation of bone marrow mesenchymal stem cells differentiation and apoptosis

摘要:

目的 探究小核仁RNA宿主基因5（SNHG5）在调控骨髓间充质干细胞成骨分化和凋亡中的作用。方法 成骨诱导液诱导骨髓间充质干细胞成骨分化，不同质粒转染分组，Western blotting法检测成骨标志物（OCN、OSX、COL1A1）的蛋白相对表达量，茜素红染色和碱性磷酸酶（ALP）检测成骨效果，实时荧光定量聚合酶链反应（qRT-PCR）检测SNHG5和OCN mRNA相对表达量。将细胞随机分为两组，实验组下调SNHG5表达，对照组不下调，同法进行成骨诱导，检测OCN、OSX及COL1A1的蛋白相对表达量，检测成骨效果，检测OCN、OSX、COL1A1及SNHG5 mRNA相对表达量。采用流式细胞术检测细胞凋亡，并检测Caspase-3活性。结果 成骨诱导后第14天的OCN、OSX及COL1A1蛋白相对表达量高于诱导后第7天和诱导前（P <0.05），诱导后第7天高于诱导前（P <0.05）。成骨诱导前、诱导后第7天、诱导后第14天的ALP活性和茜素红浓度比较，差异有统计学意义（P <0.05）。成骨诱导前、诱导后第1天、第3天、第5天、第7天、第14天的SNHG5和OCN mRNA相对表达量随时间推移逐渐升高（P <0.05）。转染不同质粒后，各组SNHG5 mRNA相对表达量比较，差异有统计学意义（P <0.05），细胞转染质粒后第14天实验组OCN、OSX及COL1A1 mRNA和蛋白相对表达量低于对照组（P <0.05），细胞转染质粒后第14天实验组ALP活性和茜素红浓度低于对照组（P <0.05），实验组细胞凋亡率和Caspase-3活性高于对照组（P <0.05）。结论 SNHG5参与骨髓间充质干细胞成骨分化和凋亡，可能在骨质疏松症的发展中起促进作用，为改善颌骨骨质疏松的治疗提供新靶点。

Abstract:

Objective To explore the role of nucleolar small RNA host gene 5 (SNHG5) in regulating osteogenic differentiation and apoptosis of bone marrow mesenchymal stem cells.Methods Osteogenic differentiation of bone marrow mesenchymal stem cells was induced by osteogenic induction solution. The protein expressions of OCN, OSX, and COL1A1 were detected by Western blotting, and the osteogenic effect was detected by alizarin red staining and alkaline phosphatase; SNHG5 were detected quantitatively by PCR; The cells were randomly divided into two groups. The expression of SNHG5 was knocked down in the experimental group and not in the control group; OCN, OSX, COL1A1, and SNHG5 were detected quantitatively by PCR; The protein expressions of OCN, OSX, and COL1A1 were detected by Western blotting, and the osteogenic effect was detected by alizarin red staining and alkaline phosphatase; Apoptosis was detected by flow cytometry; The activity of Caspase-3 was detected.Results Osteogenic induction was successful, the expressions of OCN, OSX, COL1A1, and SNHG5 increased (P < 0.05). After effectively inhibiting SNHG5, the expression of OCN, OSX, and COL1A1 in the experimental group was significantly lower than that in the control group, the activity of ALP was lower than that in the control group (P < 0.05), the degree of mineralization was lower than that in the control group (P < 0.05), the proportion of apoptosis was higher than that in the control group (P < 0.05), and the activity of Caspase-3 was higher than that in the control group (P < 0.05). The results showed that after inhibiting SNHG5 (P < 0.05), the osteogenic activity was inhibited and the apoptosis was accelerated (P < 0.05).Conclusion SNHG5 participates in the osteogenic differentiation and apoptosis of BMSCs, which may play a promoting role in the development of osteoporosis and become a new target for improving the treatment of maxillofacial osteoporosis.