Background
The regeneration of nucleus pulposus (NP) 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 NP-like cells and explored the possible mechanism of action.
Methods
The effects of ASA VI on HMSC viability and proliferation were determined by the XTT method and EDU staining. Then, Real-time qPCR, immunocytochemistry and immunofluorescence assays were used to measure the effect of ASA VI on the expression of extracellular matrix (ECM) components, such as COL2A1, aggrecan, SOX9, KRT19, PAX1, and glycosaminoglycans (GAGs), in NP cells. In addition, Western blot assay was used to measure the expression of p-ERK1/2 and p-smad2/3.
Results
ASA VI was able to promote the proliferation and differentiation of HMSCs into NP-like cells, and the optimum concentration was 1 mg/L. Western blot assay indicated that the possible mechanism might be related to the activation of p-ERK1 / 2 and p-Smad2 / 3.
Conclusions
ASA VI can promote the proliferation and differentiation of HMSCs into NP-like cells, which can potentially be used as a treatment for IVDD.
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.
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