ICU, The Second Affiliated Hospital of Shandong First Medical University, No. 366, Taishan Street, Shandong Province, 271000, Taishan, ChinaDepartment of Orthopedic, The Third Hospital of Hebei Medical University, No. 139, Ziqiang Road, Hebei Province, 050051, Shijiazhuang, ChinaDepartment of Orthopedic, The Central Hospital of Taian City, No. 29, Longtan Road, Shandong Province, 271000, Taian, ChinaDepartment of Orthopedic, The Second Affiliated Hospital of Shandong First Medical University, No. 366, Taishan Street, Shandong Province, 271000, Taishan, ChinaDoctor Student, Hebei Medical University, No. 361, Zhongshan East Road, Hebei Province, 050017, Shijiazhuang, China
Background: This study aimed to investigate the role of long non-coding RNA maternally expressed 3 and related molecular mechanisms, in osteoarthritis . Methods: Cartilage tissues of OA patients and healthy volunteers were isolated and cultured. After transfection with the appropriate constructs, chondrocytes were classified into Blank, pcDNA3.1-NC, pcDNA3.1-MEG3, si-NC, si-MEG3, pcDNA3.1-NC + mimics NC, pcDNA3.1-MEG3 + mimics NC, pcDNA3.1-NC + miR-361-5p mimics and pcDNA3.1-MEG3 + miR-361-5p ...更多
Background: This study aimed to investigate the role of long non-coding RNA maternally expressed 3 and related molecular mechanisms, in osteoarthritis . Methods: Cartilage tissues of OA patients and healthy volunteers were isolated and cultured. After transfection with the appropriate constructs, chondrocytes were classified into Blank, pcDNA3.1-NC, pcDNA3.1-MEG3, si-NC, si-MEG3, pcDNA3.1-NC + mimics NC, pcDNA3.1-MEG3 + mimics NC, pcDNA3.1-NC + miR-361-5p mimics and pcDNA3.1-MEG3 + miR-361-5p mimics groups. qRT-PCR was used to detect the expression of MEG3, miR-361-5p and FOXO1. Western blot, luciferase reporter assay, RIP, CCK-8, and flow cytometry analysis were performed to reveal the morphology, proliferation, and apoptotic status of cartilage cells. Histological analysis and immunostaining were conducted in the OA rat model. Results: Expression of MEG3 and FOXO1 was significantly decreased in OA compared with the normal group, while the expression of miR-361-5p was increased. MEG3 might serve as a ceRNA of miR-361-5p in OA chondrocytes. Moreover, using western blot analyses and the CCK-8 assay, MEG3 was shown to target miR-361-5p/FOXO1, elevate cell proliferation, and impair cell apoptosis. Functional analysis in vivo showed that MEG3 suppressed degradation of the cartilage matrix. Conclusion: MEG3 can contribute to cell proliferation and inhibit cell apoptosis and degradation of extracellular matrix via the miR-361-5p/FOXO1 axis in OA chondrocytes.收起
