Asperosaponin VI promotes human mesenchymal stem cell differentiation into nucleus pulposus like- cells via up-regulation of ERK1/2 and Smad2/3 signaling pathways.

Abstract

Background At present, the regeneration of nucleus pulposus cells is an effective method to prevent intervertebral disc degeneration (IVDD). In this study, we investigated the role of Asperosaponin VI (ASA VI), isolated from a traditional Chinese medicine (TCM), the root of Dipsacus asper Wall, in promoting human mesenchymal stem cell (HMSC) proliferation and differentiation into nucleus pulposus like-cells and explored its possible mechanism of action. Methods First, the effects of ASA VI on HMSC vitality and proliferation were determined by the XTT method and EDU staining. Then, HMSCs were cultured with ASA VI. Real-time PCR, immunocytochemistry and immunofluorescence were used to measure the expression of extracellular matrix (ECM) components in nucleus pulposus cells, such as type II collagen(COL2A1), aggrecan, SOX9, KRT19, PAX1 and glycosaminoglycans (GAGs), and Western blot was used to investigate its potential mechanism. Results ASA VI could promote HMSC differentiation into nucleus pulposus like-cells, and its optimum concentration was 1 mg/L. The Western blot assays indicated that the possible mechanisms involved upregulating the expression of P-ERK1/2 and P-Smad2/3. Conclusions ASA VI can promote HMSC proliferation and differentiation into nucleus pulposus like-cells, which can be used as a potential treatment for IVDD.