金丝桃苷上调miR-7031-5P以促进MC3T3-E1细胞的成骨分化。
Hyperin up-regulates miR-7031-5P to promote osteogenic differentiation of MC3T3-E1 cells.
作者信息
Qian Dongchen, Chen Yueyue, Qiu Xusheng, Zhu Baohua, Zhang Lin, Yan Yifeng, Chen Yixin
机构信息
Department of Orthopedic, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing, PR China.
Department of Orthopedic, The Second Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, PR China.
出版信息
Histol Histopathol. 2023 Oct;38(10):1219-1229. doi: 10.14670/HH-18-579. Epub 2022 Dec 30.
OBJECTIVE
To investigate the effects of Hyperin (Hyp) on osteogenic differentiation of MC3T3-E1 cells.
METHODS
Differentially expressed miRNA was screened by miRNA Microarray. miR-7031-5P overexpression and knockdown MC3T3-E1 cell models were constructed by transfecting miR-7031-5P mimics and inhibitor. Alizarin red staining (ARS) assay was used to observe the formation of mineralized nodules in MC3T3-E1 cells. ALP activity was detected by using ALP detection kit. Western blot assay was used to examine the changes in osteogenic differentiation-related proteins. The relationship between miR-7031-5P and Wnt7a was revealed by dual luciferase report experiments.
RESULTS
We found that miR-7031-5P was up-regulated in MC3T3-E1 cells after Hyp treatment. The results indicated that compared with the untreated group, Hyp promoted the formation of mineralized nodules and the alkaline phosphatase (ALP) activity of MC3T3-E1 cells via overexpressing miR-7031-5P. Besides, elevated miR-7031-5P increased OPN, COL1A1, and Runx2 mRNA expression. More importantly, Wnt7a was identified as the downstream target gene of miR-7031-5P promoting osteogenic differentiation of MC3T3-E1 cells.
CONCLUSIONS
Hyp up-regulated miR-7031-5P to promote osteogenic differentiation of MC3T3-E1 cells by targeting Wnt7a.
目的
探讨金丝桃苷（Hyp）对MC3T3-E1细胞成骨分化的影响。
方法
通过miRNA芯片筛选差异表达的miRNA。转染miR-7031-5P模拟物和抑制剂构建miR-7031-5P过表达和敲低的MC3T3-E1细胞模型。采用茜素红染色（ARS）试验观察MC3T3-E1细胞中矿化结节的形成。使用碱性磷酸酶（ALP）检测试剂盒检测ALP活性。采用蛋白质免疫印迹法检测成骨分化相关蛋白的变化。通过双荧光素酶报告实验揭示miR-7031-5P与Wnt7a之间的关系。
结果
我们发现Hyp处理后MC3T3-E1细胞中miR-7031-5P上调。结果表明，与未处理组相比，Hyp通过过表达miR-7031-5P促进MC3T3-E1细胞矿化结节的形成和碱性磷酸酶（ALP）活性。此外，miR-7031-5P升高增加了骨桥蛋白（OPN）、Ⅰ型胶原蛋白（COL1A1）和Runx2 mRNA的表达。更重要的是，Wnt7a被确定为miR-7031-5P促进MC3T3-E1细胞成骨分化的下游靶基因。
结论
Hyp上调miR-7031-5P，通过靶向Wnt7a促进MC3T3-E1细胞成骨分化。
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