The interaction of miR-181a-5p and sirtuin 1 regulated human bone marrow mesenchymal stem cells differentiation and apoptosis

Abstract

Osteoporosis (OP) characterizes a decrease in bone density and bone mass which leads to brittle fractures and serious damages to individuals. In recent years, various researches have proved that miRNAs act pivotally in the onset of bone-related diseases. In our research, we probed into the impact of miR-181a-5P on viability, differentiation, as well as apoptosis of human bone marrow mesenchymal stem cells (hBMSCs). Our study reported that overexpressing miR-181a-5p considerably reduced the cell growth, whereas the miR-181a-5p inhibition showed opposite results. Furthermore, the hBMSCs apoptosis percentage was visually elevated or minimized after overexpressing or silencing miR-181a-5p, respectively. Our data also indicated that miR-181a-5p overexpression significantly inhibited ALP activity, and level of OPN, Runx2 and OCN at mRNA and protein level, whereas miR-181a-5p inhibition presented opposite results. In addition, based on luciferase reporter assay, sirtuin 1 (Sirt1) was confirmed as the target of miR-181a-5p in hBMSCs. Finally, Sirt1 overexpression significantly inhibited the impact of miR-181a-5p mimic on apoptosis and inhibited differentiation, while silencing Sirt1 eliminated the inhibitory effects of miR-181a-5p on apoptosis and promoted differentiation via PI3K/AKT pathway. In conclusion, this work revealed that miR-181a-5p could regulate hBMSCs apoptosis as well as differentiation via regulating Sirt1/PI3K/AKT signaling pathway.

Salvianolic Acid B Alleviates Limb Ischemia in Mice via Promoting SIRT1/PI3K/AKT Pathway-Mediated M2 Macrophage Polarization

“…Previous reports have also shown that SIRT1 can regulate the PI3K/AKT/STAT6 pathway leading to M2 macrophage polarization (L et al 2019). Moreover, it has been demonstrated that SIRT1/PI3K/AKT pathway regulates proliferation, apoptosis, and various other pathophysiological processes as well [ 48 – 50 ]. In this study, to further investigate the underlying mechanism, we analyzed the expression of the SIRT1 and PI3K/AKT pathway.…”

Knockdown of long non-coding RNA HOTAIR promotes bone marrow mesenchymal stem cell differentiation by sponging microRNA miR-378g that inhibits nicotinamide N-methyltransferase

“…Dysregulation of osteoblast differentiation is associated with the progression of osteoporosis. Further, ALP, calcium deposition, and RUNX2 levels are often employed to illustrate osteoblast differentiation and its potential for treating OP [ 38 ]. In this study, we elucidated the effect of HOTAIR/miR-378g/NNMT on the osteogenic differentiation of BMSCs based on changes in ALP, RUNX2, and calcium deposition levels, thereby providing a potential biomarker for the treatment of OP.…”

Apremilast mitigates interleukin (IL)-13-induced inflammatory response and mucin production in human nasal epithelial cells (hNECs)

“…The total proteins were extracted from cells using the RIPA lysis buffer (50 mM Tris(pH 7.4), 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS) (Beyotime Biotechnology, Beijing, China) with the protease and phosphatase inhibitor cocktail [ 22 ], which were quantified with a bicinchoninic acid (BCA) kit (#T9300A Takara, Tokyo, Japan). Approximately 30 μg proteins were loaded and separated with 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and further transferred to the polyvinylidene fluoride (PVDF) membrane (Takara, Tokyo, Japan), and then incubated with 5% skim milk.…”