Whole Genome Expression Profiling and Signal Pathway Screening of MSCs in Ankylosing Spondylitis

Abstract

The pathogenesis of dysfunctional immunoregulation of mesenchymal stem cells (MSCs) in ankylosing spondylitis (AS) is thought to be a complex process that involves multiple genetic alterations. In this study, MSCs derived from both healthy donors and AS patients were cultured in normal media or media mimicking an inflammatory environment. Whole genome expression profiling analysis of 33,351 genes was performed and differentially expressed genes related to AS were analyzed by GO term analysis and KEGG pathway analysis. Our results showed that in normal media 676 genes were differentially expressed in AS, 354 upregulated and 322 downregulated, while in an inflammatory environment 1767 genes were differentially expressed in AS, 1230 upregulated and 537 downregulated. GO analysis showed that these genes were mainly related to cellular processes, physiological processes, biological regulation, regulation of biological processes, and binding. In addition, by KEGG pathway analysis, 14 key genes from the MAPK signaling and 8 key genes from the TLR signaling pathway were identified as differentially regulated. The results of qRT-PCR verified the expression variation of the 9 genes mentioned above. Our study found that in an inflammatory environment ankylosing spondylitis pathogenesis may be related to activation of the MAPK and TLR signaling pathways.

The long noncoding RNA GAS5 negatively regulates the adipogenic differentiation of MSCs by modulating the miR-18a/CTGF axis as a ceRNA

“…qRT-PCR was then performed with a LightCycler 480 PCR system (Roche, Basel, Switzerland) using SYBR Premix Ex Taq (TaKaRa). The detailed method was described in our previously published studies 30 . The results were first normalized to the level of GAPDH expression.…”

Abnormal inhibition of osteoclastogenesis by mesenchymal stem cells through the miR-4284/CXCL5 axis in ankylosing spondylitis

“…qRT-PCR was carried out using a LightCycler®480 PCR system (Roche) with SYBR Premix Ex Taq (TaKaRa). The steps were described in our previously published research 27 . The procedure for the qRT-PCR assay was 95 °C for 30 s and 40 cycles at 95 °C for 5 s and 60 °C for 20 s. Each sample was performed in triplicate, and the mean mRNA level was calculated.…”

Dissecting the Underlying Pharmaceutical Mechanism of Chinese Traditional Medicine Yun-Pi-Yi-Shen-Tong-Du-Tang Acting on Ankylosing Spondylitis through Systems Biology Approaches

“…Additionally, some pathways in immune system were involved in the onset and development of AS. For example, in an inflammatory environment, the activation of AMPK ( p = 5.00E-04) and TLR signaling pathway ( P = 7.30E-04) would facilitate AS incidence 30 . T cell receptor signaling pathway ( P = 9.10E-04) participated in hip joint ligament ossification 31 .…”