miR-34c-5p knockdown inhibits hypoxia induced cell apoptosis of bone marrow derived mesenchymal stem cells in rats

Abstract

Objective To explore the effect of miR-34c-5p knockdown on improving the survival rate of bone marrow derived mesenchymal stem cells (BM-MSCs). Methods The BM-MSCs were divided into normoxia group, hypoxia group, normoxia inhibitor NC group, hypoxia inhibitor NC group, normoxia microRNA(miR) inhibitor group and hypoxia miR inhibitor group. The expression level of miR-34c-5p, IGF, HGF, bFGF and VEGF was detected by real-time quantitative PCR (RT-qPCR) and ELISA. Cell apoptosis was detected by flow cytometry and TUNEL. The protein expression levels of cleaved caspase-3, -9, Bcl-2 and Bax were detected by Western blot. ATP/ADP was detected by biochemical kits, and glucose uptake was detected by flow cytometry. Results Compared with hypoxic inhibitor NC group, miR-34c-5p knockdown significantly inhibited BM-MSCs apoptosis(P＜0.01), reduced protein expression of cleaved caspase-3, -9 and Bax and increased protein expression by Bcl-2 (P＜0.01). The expression of IGF, HGF, bFGF and VEGF (P＜0.05, P＜0.01) were significantly increased; ATP/ADP rate and cell capacity of glucose transporting also increased (P＜0.05, P＜0.01). Conclusions miR-34c-5p knockdown may antagonize hypoxia induced cell apoptosis of BM-MSCs in rat.

Keywords

• mir-34c-5p knockdown|bone marrow derived mesenchymal stem cells|hypoxia|cell apoptosis