Jia Xiaolin, Yang Ming, Hu Wei, Cai San
Department of Orthopaedics, Chongqing General Hospital, Chongqing 401147, P.R. China.
Department of Orthopaedics, Chongqing Public Health Medical Center, Chongqing 400036, P.R. China.
Exp Ther Med. 2021 Jul;22(1):692. doi: 10.3892/etm.2021.10124. Epub 2021 May 2.
Osteoporosis (OP) results from an imbalance between bone formation, which is regulated by osteoblasts, and bone resorption, which is mediated by osteoclasts. MicroRNA-22-3p (miR-22-3p) expression is decreased during the process of osteoclast differentiation and p38α mitogen-activated protein kinase (MAPK)14 promotes the proliferation and differentiation of osteoclast progenitors. However, whether miR-22-3p could target MAPK14 to regulate the progression of OP remains unknown, which was the aim of the present study. CD14 PBMCs were used for the establishment of osteoclastic differentiation . In the present study, reverse transcription quantitative PCR was used to determine the mRNA expression of MAPK14, tartrate resistant acid phosphatase (TRAP), nuclear factor of activated T-cells (NFATC1) and cathepsin K (CTSK). Western blotting was applied to determine the protein expression of MAPK14, TRAP, NFATC1, CTSK, p-p65 and p65. Dual luciferase reporter assay was applied to confirm the relation between miR-22-3p and MAPK14. Cell Counting Kit-8 assay and flow cytometry assays were used to determine the cell proliferation and cell apoptosis, respectively. The results demonstrated that miR-22-3p expression was lower while MAPK14 expression was higher in the serum from patients with OP compared with healthy volunteers. Furthermore, miR-22-3p expression was negatively correlated with MAPK14 expression in patients with OP. In addition, miR-22-3p expression was decreased and MAPK14 expression was increased during the progression of CD14peripheral blood mononuclear cells (PBMCs) osteoclastic differentiation in a time-dependent manner. Furthermore, miR-22-3p inhibited the proliferation and differentiation and promoted the apoptosis of CD14PBMCs by targeting MAPK14. In summary, the findings from the present study suggested that miR-22-3p may serve a potential therapeutic role in patients with OP.
骨质疏松症(OP)是由成骨细胞调节的骨形成与破骨细胞介导的骨吸收之间的失衡所致。在破骨细胞分化过程中,微小RNA-22-3p(miR-22-3p)表达降低,且p38α丝裂原活化蛋白激酶(MAPK)14促进破骨细胞祖细胞的增殖和分化。然而,miR-22-3p是否可靶向MAPK14以调节OP的进展仍不清楚,这也是本研究的目的。采用CD14外周血单个核细胞(PBMC)建立破骨细胞分化模型。在本研究中,采用逆转录定量PCR检测MAPK14、抗酒石酸酸性磷酸酶(TRAP)、活化T细胞核因子(NFATC1)和组织蛋白酶K(CTSK)的mRNA表达。应用蛋白质印迹法检测MAPK14、TRAP、NFATC1、CTSK、磷酸化p65和p65的蛋白表达。采用双荧光素酶报告基因检测法证实miR-22-3p与MAPK14之间的关系。分别采用细胞计数试剂盒-8法和流式细胞术检测细胞增殖和细胞凋亡。结果表明,与健康志愿者相比,OP患者血清中miR-22-3p表达较低,而MAPK14表达较高。此外,OP患者中miR-22-3p表达与MAPK14表达呈负相关。另外,在CD14外周血单个核细胞(PBMC)破骨细胞分化过程中,miR-22-3p表达呈时间依赖性降低,MAPK14表达呈时间依赖性增加。此外,miR-22-3p通过靶向MAPK14抑制CD14PBMCs的增殖和分化并促进其凋亡。总之,本研究结果提示miR-22-3p可能对OP患者具有潜在治疗作用。