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

Abstract

Objective: 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 [CM(155mm]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.