Mus musculus
Protocols: Synovial tissues were harvested from all experimental groups (control group and the three treatment groups). Immediately after collection, TRIzol reagent was added directly to the samples, which were then stored at −80°C until RNA extraction. Total RNA was extracted using the TRIzol reagent according to the Qiagen manufacturer's protocol (all steps were respected, and DNase treatment was included to eliminate residual DNA).  RNA quality was assessed prior to sequencing, and only RNA samples with an RNA Integrity Number (RIN) greater than 7 were included for downstream analysis. RNA samples were subsequently sent to BGI Informatics (Hong Kong) for RNA-sequencing library preparation. Sequencing libraries were prepared from total RNA using BGI's standard RNA-sequencing library preparation workflow. Libraries were purified and subjected to quality assessment prior to sequencing.  mRNA was enriched from total RNA using oligo(dT)-attached magnetic beads. The enriched poly(A)+ mRNA was fragmented using a fragmentation buffer, followed by reverse transcription with random N6 primers to generate double-stranded cDNA. The synthesized double-stranded cDNA was end-repaired and 5′-phosphorylated, and an adenine (A) overhang was added to the 3′ ends. A bubble-shaped adapter with a 3′ thymine (T) overhang was then ligated to the cDNA fragments. The ligation products were PCR-amplified using specific primers. The PCR products were denatured into single strands, and single-stranded circular DNA libraries were generated using a bridging primer. The constructed libraries underwent quality control before high-throughput sequencing.