miR-146a 调控创伤性股骨头坏死骨髓基质细胞的成骨分化和增殖。
MiR-146a regulates osteogenic differentiation and proliferation of bone marrow stromal cells in traumatic femoral head necrosis.
机构信息
Department of Bone and Joint Surgery, Affiliated Hospital of Jining Medical University, Jining, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Jan;23(2):441-448. doi: 10.26355/eurrev_201901_16853.
OBJECTIVE
To investigate the regulatory mechanism of micro ribonucleic acid (miR)-146a in osteogenic differentiation and proliferation of bone marrow stromal cells (BMSCs) in traumatic femoral head necrosis.
PATIENTS AND METHODS
Femoral neck fracture patients undergoing surgery were divided into necrosis group and non-necrosis group. The expression level of miR-146a in BMSCs isolated from these patients was detected via quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The clinical correlation of miR-146a with BMSCs in traumatic femoral head necrosis was explored. The regulatory effects of miR-146a on osteogenic differentiation and proliferation of BMSCs in traumatic femoral head necrosis were detected. Moreover, the cell proliferation was analyzed via cell counting kit-8 (CCK-8) assay. The deposition of calcium on the cell surface was detected via alizarin red staining to evaluate the osteogenic differentiation. The messenger RNA (mRNA) expressions of osteogenesis-specific genes, alkaline phosphatase (ALP), and osteocalcin (Ocn) in BMSCs undergoing osteogenic differentiation were detected via qRT-PCR.
RESULTS
Expression level of miR-146a in BMSCs of necrosis group was significantly lower than that in non-necrosis group, and the difference was statistically significant (p<0.01). CCK-8 assay revealed that the proliferation of BMSCs was significantly enhanced in miR-146a-mimic group compared with that in miR-NC group, while it significantly declined in miR-146a-inhibitor group compared with that in miR-NC group. The results of alizarin red staining showed that the deposition of calcium obviously increased in miR-146a-mimic group compared with that in miR-NC group, indicating that the osteogenic differentiation ability is significantly enhanced, while it markedly decreased in miR-146a-inhibitor group compared with that in miR-NC group. The detection of osteogenesis-specific genes via qRT-PCR manifested that the mRNA expressions of ALP and Ocn remarkably increased in miR-146a-mimic group compared with those in miR-NC group, and there were statistically significant differences (p<0.05). The mRNA expressions of ALP and Ocn remarkably decreased in miR-146a-inhibitor group compared with those in miR-NC group, and there were statistically significant differences (p<0.05), suggesting the inhibited osteogenic differentiation ability.
CONCLUSIONS
We showed that miR-146a regulates the osteogenic differentiation and proliferation of BMSCs in traumatic femoral head necrosis.
目的
探讨微小核糖核酸-146a（miR-146a）在创伤性股骨头坏死骨髓基质细胞（BMSCs）成骨分化和增殖中的调控机制。
方法
将接受手术的股骨颈骨折患者分为坏死组和非坏死组。通过实时定量聚合酶链反应（qRT-PCR）检测从这些患者中分离的 BMSCs 中 miR-146a 的表达水平。探讨 miR-146a 与创伤性股骨头坏死中 BMSCs 的临床相关性。检测 miR-146a 对创伤性股骨头坏死中 BMSCs 成骨分化和增殖的调控作用。此外，通过细胞计数试剂盒-8（CCK-8）检测细胞增殖。通过茜素红染色检测细胞表面钙沉积，评估成骨分化。通过 qRT-PCR 检测成骨分化过程中 BMSCs 中骨形成特异性基因碱性磷酸酶（ALP）和骨钙素（Ocn）的 mRNA 表达。
结果
坏死组 BMSCs 中 miR-146a 的表达水平明显低于非坏死组，差异具有统计学意义（p<0.01）。CCK-8 检测显示，miR-146a 模拟物组 BMSCs 的增殖明显高于 miR-NC 组，而 miR-146a 抑制剂组 BMSCs 的增殖明显低于 miR-NC 组。茜素红染色结果表明，miR-146a 模拟物组钙沉积明显多于 miR-NC 组，表明成骨分化能力明显增强，而 miR-146a 抑制剂组则明显少于 miR-NC 组。通过 qRT-PCR 检测骨形成特异性基因发现，miR-146a 模拟物组 ALP 和 Ocn 的 mRNA 表达明显高于 miR-NC 组，差异具有统计学意义（p<0.05）。miR-146a 抑制剂组 ALP 和 Ocn 的 mRNA 表达明显低于 miR-NC 组，差异具有统计学意义（p<0.05），表明成骨分化能力受到抑制。
结论
我们表明，miR-146a 调节创伤性股骨头坏死中 BMSCs 的成骨分化和增殖。
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