Adipose-derived Stem Cells Induce Autophagic Activation and Inhibit Catabolic Response to Pro-Inflammatory Cytokines in Rat Chondrocytes.

Objective: Adipose-derived stem cells (ADSCs) have been demonstrated to have an anti-apoptosis effect on chondrocytes; However, their effect on autophagic activation remains unclear. We sought to explore whether ADSCs can activate autophagy and inhibit IL-1 beta- and lipopolysaccharide (LPS)-induced catabolism in chondrocytes.Methods: ADSCs and chondrocytes were collected from SD rats. The biologic characteristics of ADSCs were analyzed by flow cytometric analysis, Oil red O and Alizarin Red staining. Autophagic level and autophagic flux were revealed by Western blotting for LC3-II and SQSTM1/P62, MDC (monodansylcadaverine) staining and mRFP-GFP-LC3 analysis. The mTOR pathway was investigated by Western blotting for p-mTOR. The mRNA level of matrix metalloproteinases (MMPs) and thrombospondin motifs (ADAMTSs) was detected by real-time PCR.Results: The typical surface markers and differentiation potentials of ADSCs were proved. ADSCs enhanced the expression of LC3-II/LC3-I and reduced SQSTM1 levels in IL-1 beta-induced chondrocytes after 24 and 48 h co-culturing and in LPS-induced chondrocytes after 48 h co-culturing respectively. mRFP-GFP-LC3 analysis suggested that autophagosomes and autolysosomes were formed earlier in IL-1 beta-treated chondrocytes than in LPS-treated chondrocytes. Bafilomycin A1 treatment further increased the LC3-II/LC3-I level in chondrocytes in co-culture with ADSCs. The mTOR pathway was inhibited in the chondrocytes in co-culture with ADSCs. Finally, ADSCs inhibited the increase of MMPs and ADAMTSs in chondrocytes induced by IL-1 beta and LPS.Conclusions: ADSCs seem able to activate autophagy and inhibit catabolism in chondrocytes in an inflammation environment, and the mTOR pathway might be involved in the autophagy activation. (C) 2016 Published by Elsevier Ltd on behalf of Osteoarthritis Research Society International.