PDF(Pubmed)
Abstract：
BACKGROUND: The progression of osteoporosis (OP) can dramatically increase the risk of fractures, which seriously disturb the life of elderly individuals. Specific protein 1 (SP1) is involved in OP progression. However, the mechanism by which SP1 regulates OP progression remains unclear.
OBJECTIVE: This study investigated the mechanism underlying the function of SP1 in OP.
METHODS: SAMP6 mice were used to establish an in vivo model of age-dependent OP, and BALB/c mice were used as controls. BMSCs were extracted from two subtypes of mice. Hematoxylin and eosin staining were performed to mark the intramedullary trabecular bone structure to evaluate histological changes. ChIP assay was used to assess the targeted regulation between SP1 and miR-133a-3p. The binding sites between MAPK3 and miR-133a-3p were verified using a dual-luciferase reporter assay. The mRNA levels of miR-133a-3p and MAPK3 were detected using quantitative reverse transcription polymerase chain reaction (RT-qPCR). The protein expression of SP1, MAPK3, Colla1, OCN, and Runx2 was examined using Western blotting. Alkaline phosphatase (ALP) kit and Alizarin Red S staining were used to investigate ALP activity and mineralized nodules, respectively.
RESULTS: The levels of SP1 and miR-133a-3p were upregulated, whereas the expression of MAPK3 was downregulated in BMSCs from SAMP6 mice, and miR-133a-3p inhibitor accelerated osteogenic differentiation in BMSCs. SP1 directly targeted miR-133a-3p, and MAPK3 was the downstream mRNA of miR-133a-3p. Mechanically, SP1 accelerated osteogenic differentiation in BMSCs via transcriptional mediation of the miR-133a-3p/MAPK3 axis.
CONCLUSIONS: SP1 regulates osteogenic differentiation by mediating the miR-133a-3p/MAPK3 axis, which would shed new light on strategies for treating senile OP.
OBJECTIVE: This study investigated the mechanism underlying the function of SP1 in OP.
METHODS: SAMP6 mice were used to establish an in vivo model of age-dependent OP, and BALB/c mice were used as controls. BMSCs were extracted from two subtypes of mice. Hematoxylin and eosin staining were performed to mark the intramedullary trabecular bone structure to evaluate histological changes. ChIP assay was used to assess the targeted regulation between SP1 and miR-133a-3p. The binding sites between MAPK3 and miR-133a-3p were verified using a dual-luciferase reporter assay. The mRNA levels of miR-133a-3p and MAPK3 were detected using quantitative reverse transcription polymerase chain reaction (RT-qPCR). The protein expression of SP1, MAPK3, Colla1, OCN, and Runx2 was examined using Western blotting. Alkaline phosphatase (ALP) kit and Alizarin Red S staining were used to investigate ALP activity and mineralized nodules, respectively.
RESULTS: The levels of SP1 and miR-133a-3p were upregulated, whereas the expression of MAPK3 was downregulated in BMSCs from SAMP6 mice, and miR-133a-3p inhibitor accelerated osteogenic differentiation in BMSCs. SP1 directly targeted miR-133a-3p, and MAPK3 was the downstream mRNA of miR-133a-3p. Mechanically, SP1 accelerated osteogenic differentiation in BMSCs via transcriptional mediation of the miR-133a-3p/MAPK3 axis.
CONCLUSIONS: SP1 regulates osteogenic differentiation by mediating the miR-133a-3p/MAPK3 axis, which would shed new light on strategies for treating senile OP.
摘要：
背景：骨质疏松症（OP）的进展可以显着增加骨折的风险，这严重扰乱了老年人的生活。特异性蛋白1(SP1)参与OP进程。然而,SP1调节OP进展的机制尚不清楚.
目的：本研究探讨了SP1在OP中的作用机制。
方法：用SAMP6小鼠建立年龄依赖性OP的体内模型，和BALB/c小鼠用作对照。从两种亚型小鼠中提取BMSCs。进行苏木精和伊红染色以标记髓内骨小梁结构以评估组织学变化。ChIP测定用于评估SP1和miR-133a-3p之间的靶向调节。MAPK3和miR-133a-3p之间的结合位点使用双荧光素酶报告物测定来验证。使用定量逆转录聚合酶链反应(RT-qPCR)检测miR-133a-3p和MAPK3的mRNA水平。SP1、MAPK3、Colla1、OCN、使用Western印迹检查Runx2。碱性磷酸酶（ALP）试剂盒和茜素红S染色用于研究ALP活性和矿化结节，分别。
结果：SP1和miR-133a-3p水平上调，而SAMP6小鼠的BMSCs中MAPK3的表达下调，miR-133a-3p抑制剂可加速BMSCs的成骨分化。SP1直接靶向miR-133a-3p，MAPK3是miR-133a-3p的下游mRNA。机械上,SP1通过miR-133a-3p/MAPK3轴的转录介导加速BMSCs成骨分化。
结论：SP1通过介导miR-133a-3p/MAPK3轴调节成骨分化，这将为治疗老年OP的策略提供新的思路。
目的：本研究探讨了SP1在OP中的作用机制。
方法：用SAMP6小鼠建立年龄依赖性OP的体内模型，和BALB/c小鼠用作对照。从两种亚型小鼠中提取BMSCs。进行苏木精和伊红染色以标记髓内骨小梁结构以评估组织学变化。ChIP测定用于评估SP1和miR-133a-3p之间的靶向调节。MAPK3和miR-133a-3p之间的结合位点使用双荧光素酶报告物测定来验证。使用定量逆转录聚合酶链反应(RT-qPCR)检测miR-133a-3p和MAPK3的mRNA水平。SP1、MAPK3、Colla1、OCN、使用Western印迹检查Runx2。碱性磷酸酶（ALP）试剂盒和茜素红S染色用于研究ALP活性和矿化结节，分别。
结果：SP1和miR-133a-3p水平上调，而SAMP6小鼠的BMSCs中MAPK3的表达下调，miR-133a-3p抑制剂可加速BMSCs的成骨分化。SP1直接靶向miR-133a-3p，MAPK3是miR-133a-3p的下游mRNA。机械上,SP1通过miR-133a-3p/MAPK3轴的转录介导加速BMSCs成骨分化。
结论：SP1通过介导miR-133a-3p/MAPK3轴调节成骨分化，这将为治疗老年OP的策略提供新的思路。
