1. Icariin regulates miR-23a-3p-mediated osteogenic differentiation of BMSCs via BMP-2/Smad5/Runx2 and WNT/β-catenin pathways in osteonecrosis of the femoral head
- Author
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Jian-zhao Liao, Song Li, Ding-peng Chen, Yue-ping Chen, Feng Chen, Yuan Chai, Yong-qian Liu, Bin Gan, Xiao-yun Zhang, and Hua-nan Li
- Subjects
TRACP-5b, tartrate-resistant acid phosphatase 5b ,ONFH, osteonecrosis of the femoral head ,RT-PCR, Real time PCR ,BMP-4, bone morphogenetic protein-4 ,Pharmaceutical Science ,BAP, bone-specific alkaline phosphatase ,RM1-950 ,Bone morphogenetic protein 2 ,DMEM, Dulbecco’s modified Eagle’s medium ,chemistry.chemical_compound ,Femoral head ,FBS, fetal bovine serum ,stomatognathic system ,medicine ,HE, Hematoxylin‐eosin ,BMSCs, bone marrow-derived mesenchymal stem cells ,Pharmacology ,WNT/β-catenin pathway ,BMP-2/Smad5/Runx2 pathway ,Wnt signaling pathway ,CTX-1, C-terminal telopeptides of type I collagen ,DR, Digital Radiography ,BMP-2, bone morphogenetic protein-2 ,Icariin ,RUNX2 ,medicine.anatomical_structure ,miR-23a-3p ,chemistry ,Osteonecrosis of the femoral head ,Catenin ,NTX-1, N-terminal telopeptides of type I collagen ,SI, icariin-containing serum ,embryonic structures ,Cancer research ,LPS, lipopolysaccharide ,Original Article ,Therapeutics. Pharmacology ,OC, osteocalcin - Abstract
Icariin is commonly used for the clinical treatment of osteonecrosis of the femoral head (ONFH). miR-23a-3p plays a vital role in regulating the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). The present study aimed to investigate the roles of icariin and miR-23a-3p in the osteogenic differentiation of BMSCs and an ONFH model. BMSCs were isolated and cultured in vitro using icariin-containing serum at various concentrations, and BMSCs were also transfected with a miR-23a inhibitor. The alkaline phosphatase (ALP) activity and cell viability as well as BMP-2/Smad5/Runx2 and WNT/β-catenin pathway-related mRNA and protein expression were measured in BMSCs. Additionally, a dual-luciferase reporter assay and pathway inhibitors were used to verify the relationship of icariin treatment/miR-23a and the above pathways. An ONFH rat model was established in vivo, and a 28-day gavage treatment and lentivirus transfection of miR-23a-3p inhibitor were performed. Then, bone biochemical markers (ELISA kits) in serum, femoral head (HE staining and Digital Radiography, DR) and the above pathway-related proteins were detected. Our results revealed that icariin treatment/miR-23a knockdown promoted BMSC viability and osteogenic differentiation as well as increased the mRNA and protein expression of BMP-2, BMP-4, Runx2, p-Smad5, Wnt1 and β-catenin in BMSCs and ONFH model rats. In addition, icariin treatment/miR-23a knockdown increased bone biochemical markers (ACP-5, BAP, NTXI, CTXI and OC) and improved ONFH in ONFH model rats. In addition, a dual-luciferase reporter assay verified that Runx2 was a direct target of miR-23a-3p. These data indicated that icariin promotes BMSC viability and osteogenic differentiation as well as improves ONFH by decreasing miR-23a-3p levels and regulating the BMP-2/Smad5/Runx2 and WNT/β-catenin pathways.
- Published
- 2021