Effects of Shujin Jiegu Decoction on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells by regulating miR-30a-5p/Sprouty1 gene

Abstract

Objective To investigate the effects of Shujin Jiegu Decoction on the proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) and its possible mechanism. Methods hMSCs were cultured in vitro. Methylthiazole tetrazolium (MTT) colorimetric assay was used to detect the effects of Shujin Jiegu Decoction and Sprouty1 down-regulation at different levels on the proliferation of hMSCs. Real-time quantitative PCR (qRT-PCR) was used to detect the expression of micro RNA-30a-5p (miR-30a-5p) in hMSCs, and RT-qPCR and western blot were used to measure the mRNA and protein expression levels of Sprouty1, Runt-related transcription factor 2 (RUNX2) , and osteoblast-specific transcription factor Osterix (OSX) in different hMSCs groups. Dual luciferase reporter assay was used to validate the role of miR-30a-5p in targeted regulation of Sprouty1. Results Shujin Jiegu Decoction was shown to promote the proliferation of hMSCs in a concentration-dependent manner, exhibiting the most prominent effect at a level of 1 mg/ml. Compared with the control group, Shujin Jiegu Decoction could significantly promote the mRNA and protein expression of miR30a-5p, RUNX2 and OSX (P<0.05) , and inhibit the mRNA and protein expression of Sprouty1 (P<0.05) , in hMSCs. The results of dual luciferase reporter gene assay showed that miR-30a-5p negatively target-regulated the expression of Sprouty1. Down-regulation of Sprouty1 promoted hMSCs proliferation and expression of OSX and RUNX2 mRNA and protein in hMSCs. Conclusion Shujin Jiegu Decoction may promote the proliferation and osteogenic differentiation of hMSCs by regulating the expression of miR-30a-5p/Sprouty1 gene. Key words: Bone marrow mesenchymal stem cells; Shujinjiegu Decoction; miR-30a-5p; Sprouty1