A ROS/GAS5/SIRT1 reinforcing feedback promotes oxidative stress-induced adipogenesis in bone marrow-derived mesenchymal stem cells during osteoporosis

LincGAS5 have been reported to regulate the progression of osteoporosis (OP). However, the relationship between LincGAS5 and reactive oxygen species (ROS) in osteoporosis were still unclear.Bilateral ovariectomy (OVX) rat were established as OP model and verified by the Micro-computed tomography. The ROS level of BMSCs derived from OVX and control rat were detected by Immunofluorescence (IF) and flow cytometry. The role of GAS5, miR-23b-3p and SIRT1 on the osteogenic differentiation were dectected by ARS saining and ALP staining, while the The Oil Red O staining and flow cytometry (FCM) were hired to determine adipogenic differentiation of BMSCs under different treatment. The expression of GAS5,miR-23b-3p and SIRT1 in BMSCs was detected by RT-qPCR and the correlation among them was analyzed. In addition, Luciferase activity was used to detect whether miR-23b-3p combined with GAS5 and SIRT1 in OP mice BMSCs.We established the OVX rat model and found higher ROS level in BMSCs isolated from OVX rats. Meanwhile, GAS5 was down-regulated by ROS and remarkably lowly expressed in OVX rat comparing with the negative control. We confirmed GAS5 inhibited adipogenesis and promoted osteoporosis progression. Mechanically, GAS5 bound with miR-23b-3p and suppressed its biological function. We also identified that miR-23b-3p bound with Sirtuin 1 (SIRT1) and decreased its stability. Furthermore, SIRT1 suppressed ROS production in BMSCs, which in turn un-regulated GAS5 expression through ROS-GAS5 axis.We identified a negative feedback loop, ROS-GAS5-SIRT1, in osteoporosis progression. Our findings provided potential targets and biomarkers for osteoporosis prevention and treatment.