RESUMEN
In this study, we aimed to identify the specific microRNAs (miRNAs) that are involved in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) from ovariectomized (OVX) mice, and to further explore the mechanism by which these miRNAs regulate osteogenic differentiation. Based on the existing studies, the expression of seven miRNAs in BMSCs from OVX mice was evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of miR-133a-3p and osteogenesis-related genes (runt-related transcription factor 2 (Runx2), Osterix, alkaline phosphatase (ALP), and osteopontin) in BMSCs treated with miR-133a-3p mimics or inhibitors was detected by qRT-PCR or Western blotting. Osteogenesis efficiency was determined using ALP and alizarin red staining. The effector-target relationship between miR-133a-3p and ankyrin repeat domain 44 (ANKRD44) was confirmed by bioinformatics and a dual luciferase assay. Among the seven selected miRNAs, miR-133a-3p expression was significantly increased in BMSCs from OVX mice. Overexpression of miR-133a-3p dramatically inhibited the expression of osteogenesis-related genes in BMSCs and reduced ALP activity and mineralization. However, these processes were markedly ameliorated upon miR-133a-3p inhibition. Moreover, miR-133a-3p appeared to target ANKRD44, and the ANKRD44 expression was negatively regulated by miR-133a- 3p. Furthermore, ANKRD44 upregulation eliminated the anti-osteogenic differentiation effects of miR-133a-3p in BMSCs. Thus, our results indicated that miR-133a-3p inhibits the osteogenic differentiation of BMSCs by suppressing ANKRD44.
Asunto(s)
Células Madre Mesenquimatosas , MicroARNs , Animales , Repetición de Anquirina , Células de la Médula Ósea , Diferenciación Celular , Células Cultivadas , Ratones , MicroARNs/genética , Osteogénesis/genéticaRESUMEN
BACKGROUND: This study is a retrospective case analysis of 143 patients who suffered from severe intracranial hemorrhage and underwent a fast and simple procedure of cranial drilling followed with external ventricle drain treatment (referred as Fast-D here after) during 2003-2013 to evaluate the clinical effectiveness of the treatment. METHODS: Fast-D procedure was conducted on 143 patients with severe acute craniocerebral diseases. Those patients were evaluated using activities of daily living (ADL) scales at hospital discharge and after 6-month of physical therapy, and were compared to 36 patients with similar craniocerebral diseases but received the traditional Dandy's surgical treatment. RESULTS: At discharge, 11% (16 cases) was classified as ADL I (fully functional for physical and social activities); 26% (37 cases) had ADL II (fully functional for physical activities but partially impaired for social activities); 34% (49 cases) was ADL III (require assistance performing physical activities); 9% (13 cases) was ADL IV (being conscious, but completely lost ability of physical activities); 27% (10 cases) was ADL V (vegetative stage); and 13% (18 cased) was ADL VI (died) among the 143 patients. Six-month physical therapy improved ADL in 88% of the patients. Those outcomes are equal or better than the more complicated Dandy's procedure probably due to the time-saving factor. CONCLUSION: Fast-D procedure is much faster (6.7 min vs. 53.6 min of the Dandy's procedure) and can be performed outside operating rooms (computed tomography room or bedside). This technique could serve as a tool to rapidly release intracranial pressure and reduce subsequent morbidity and mortality of severe craniocerebral diseases when resource and condition are limited and more elaborate operating room procedures are not possible.