Effects of Bisphosphonates on Beclin1 and LC3Ⅱ Induced by High-glucose in Rat Bone Marrow Mesenchymal Stem Cells

Abstract

To investigate the effects of bisphosphonates on autophagy induced by high-glucose in rat bone marrow mesenchymal stem cells (BMSCs). Methods BMSCs were isolated and cultured in vitro,identified by undergoing osteogenic/chondrogenic/adipogenic differentiation, the concentration of bisphosphonates was determined by CCK-8 method. The cells were cultured in normal glucose (5.6 mmol/L D-glucose), high glucose (30 mmol/L D-glucose), and high glucose with bisphosphonates (30 mmol/L D-glucose+10-9 mmol/L bisphosphonates). At 48 h, mRNA expression levels of autophagy related genes Beclin1 and microtubule-associated protein 1 light chain 3 （LC3） were dected by real-time PCR, protein expression levels of Beclin1 and LC3Ⅱ were detected by Western blot, and the autophagy body was observed by transmission electron microscopy (TEM). Results The results showed that BMSCs had the ability of osteogenenic, chondrogenic and adipogenic differentiation. Compared with the control group and high glucose with bisphosphonates group, the mRNA expressions of Beclin1 and LC3 and protein expressions of Beclin1 and LC3Ⅱ in the high glucose group were increased (P<0.01 or P<0.05). TEM showed that the number of autophagy body in high glucose group was higher than that in normal group and high glucose with bisphosphonates group. Conclusion Bisphosphonates may play a role of down-regulating the expression of Beclin1 and LC3Ⅱ induced by high-glucose in BMSCs.

Keywords: Autophagy, Bisphosphonates, Diabetic osteoporosis