Cordycepin Protected Against The TNF-α-induced Inhibition Of Osteogenic Differentiation Of Human Adipose-derived Mesenchymal Stem Cells

Background and objectiveCordycepin,3’-deoxyadenosine,is an effective component isolated from the rare Chinese caterpillar fungus Cordyceps militaris.It exerts potent anti-inflammatory actions in different cell and animal models.However,its action remains unclear on the TNF-α-induced inhibition of osteogenic differentiation of adipose-derived mesenchymal stem cells(ADMSCs).MethodsHuman ADMSCs isolated from patients’ abdominal subcutaneous fats were cultured with different concentration of TNF-α and cordycepin in osteogenesis medium.ADMSCs proliferation and osteogenesis were evaluated by ALP staining,ARS staining,RT-PCR and Western blotting.ResultsIn the present study,we demonstrated that cordycepin induced cell death at 20 and 40μg/mL.Interestingly,10 μg/mL cordycepin abrogated the cell death induced by 20 ng/mL TNF-α.Meanwhile,cordycepin exhibited a dosedependent regulation of the osteogenesis of human ADMSCs:it promoted the differentiation at 10 μg/mL,whereas inhibited differentiation at 40 μg/mL.Furthermore,we discovered that 10μg/mL cordycepin protected against the TNF-α(induced inhibition of osteogenic differentiation of human ADMSCs.It was also revealed that 10 μg/mL cordycepin restored Runx2 and Osx mRNA levels,which were significantly inhibited by TNF-aduring osteogenesis.At the same time,we found that 10 μg/mL cordycepin suppressed TNF-a-activated NF-κB signaling,by inhibiting IκBα phosphorylation and subsequent p65 release and translocation into the cell nucleus.ConclusionsOf clinical interest,the present study revealed mechanisms involved in inflammatory cytokine-inhibited osteogenesis,and it highlights the potential of cordycepin to promote the osteogenesis of human ADMSCs in cell-based therapy for inflammatory bone diseases.