Prediction and expression of microrna targeting interleukin-17a in chondrocytes and synovial fibroblasts isolated from osteoarthritic knee
Osteoarthritis (OA) is a prevalent degenerative joint disorder associated with pain and disability. Dysregulation of microRNAs (miRNAs) and inflammation processes play significant roles in OA pathogenesis, with interleukin-17A (IL-17A) as the key proinflammatory cytokine implicated in the cartila...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2023
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| Subjects: | |
| Online Access: | http://eprints.usm.my/60269/1/Ng%20Jun%20Quan-E.pdf |
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| Summary: | Osteoarthritis (OA) is a prevalent degenerative joint disorder associated with pain and
disability. Dysregulation of microRNAs (miRNAs) and inflammation processes play
significant roles in OA pathogenesis, with interleukin-17A (IL-17A) as the key proinflammatory
cytokine implicated in the cartilage degradation and synovial
inflammation. Understanding the regulation of IL-17A signalling in chondrocytes and
synovial fibroblasts are crucial for unravelling the underlying disease mechanisms.
This study aimed to predict and determine the expression level of miRNAs targeting
IL-17A in chondrocytes and synovial fibroblasts isolated from osteoarthritic knee
joints. Bioinformatics tools, including TargetScan, miRWalk, and miRDB were
employed to identify potential miRNAs targeting IL-17A. The miRNA with the
highest and the lowest predicted binding affinity were selected. DIANA-mirPATH
analysis web server was used to identify potential pathways that could be targeted by
the selected miRNAs. The expression levels of these miRNAs were assessed via
quantitative polymerase chain reaction (qPCR) in isolated chondrocytes and synovial
fibroblasts from OA patients. ΔCT value was calculated in the study. Hsa-mir-1913
exhibited the highest predicted binding affinity to IL-17A mRNA, while hsa-mir-514a-
5p showed lower binding affinity. These miRNAs were used as a comparative measure
to validate the accuracy of the results obtained from the bioinformatics tools. Target
prediction of the expressed miRNAs identified different inflammation-linked pathways that are consistent with the previous studies. The qPCR analysis
demonstrated that hsa-mir-1913 showed consistent high expression in chondrocytes
and synovium fibroblasts samples (14.36 ± 0.72; 14.12 ± 0.69). On the other hand,
hsa-mir-514a-5p (-4.65 ± 0.45; -5.05 ± 0.60) exhibited lower expression in both
samples. It is essential to acknowledge that this study represents a preliminary
investigation into miRNAs that potentially regulate IL-17A in chondrocytes and
synovial fibroblasts of osteoarthritis patients. Further research is needed to fully
understand the specific mechanisms and functional consequences of these miRNAs in
the pathogenesis of osteoarthritis. This study successfully predicted and detected the
expression of miRNA targeting IL-17A in chondrocytes and synovial fibroblasts
isolated from osteoarthritic knee joints. The identification of miRNA targeting IL-17A
may highlight the intricate role of miRNAs in modulating the inflammatory and
degenerative process associated with OA and open new avenues for future research
and development of targeted therapies for OA. |
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