TNF-α Induced the Enhanced Apoptosis of Mesenchymal Stem Cells in Ankylosing Spondylitis by Overexpressing TRAIL-R2

Figure 3

ASMSCs had increased protein expression of cleaved caspase-3, TRAIL-R2, and FADD and increased gene expression of TRAIL-R2 and FADD during apoptosis. (a) The Proteome Profiler™ Human Apoptosis Array Kit was used to determine the expression levels of 35 apoptosis-related proteins in HDMSCs () and ASMSCs () after treatment with TNF-α/CHX. The intensity of each pair of spots represents repeated measures of one protein. ASMSCs had lower pro-caspase-3 (a.1) expression but higher cleaved caspase-3 (a.2), TRAIL-R2 (a.3), and FADD (a.4) expression levels than HDMSCs. No significant differences in the expression of the remaining 31 proteins were found between HDMSCs and ASMSCs (b) qRT-PCR was performed to confirm the results of the Proteome Profiler™ Human Apoptosis Array. The results of TRAIL-R2 and FADD in HDMSCs () and ASMSCs () were consistent with those of the Proteome Profiler™ Human Apoptosis Array. However, total caspase-3 mRNA expression was not different between HDMSCs and ASMSCs. (c) Western blot analysis was performed on proteins isolated from HDMSCs () and ASMSCs () to further confirm the results of the Proteome Profiler™ Human Apoptosis Array, and the results of both methods were in agreement. Values are presented as the mean ± SD. indicates between HDMSCs and ASMSCs.