An analysis of Treg/Th17 cells imbalance associated microRNA networks regulated by moxibustion therapy on Zusanli (ST36) and Shenshu (BL23) in mice with collagen induced arthritis

Moxibustion is an emerging alternative therapy for rheumatoid arthritis (RA) in the eastern world, especially in China. However, the mechanism underlying this condition has not yet been elucidated. This study aimed to explore how moxibustion reduced arthritis by regulating Treg/Th17 cell imbalance and the role of differentiation-associated keynote microRNAs (miRNAs). Moxibustion therapy was administered to mice with collagen-induced arthritis (CIA). The arthritis index, histopathological changes, inflammatory factors, and Treg/Th17 cell balance were analyzed. MiRNAs from CD4(+) T cells were analyzed based on the RNA-seq technology. Treg/Th17 cell differentiation-related miRNAs and their target genes were identified from the online database of miRDB, TargetScan, and miRTarBase. The expression of miRNAs and their target genes was verified by quantitative reverse transcription polymerase chain reaction and Western blot analysis. The binding sites of miRNAs and target genes were predicted by miRDB, and the targeting relationship between them was verified by dual-luciferase reporter assay. Arthritis in mice with CIA was reduced by moxibustion therapy, and Treg/Th17 cell imbalance was regulated. Seventeen upregulated and twenty-three downregulated miRNAs were identified in moxibustion-treated mice. Seven upregulated miRNAs, seven downregulated miRNAs, and five Treg/Th17 cell differentiation-associated target genes were predicted. Among them, miR-144-3p and Hif1a were suggested to be the keynote miRNA and target gene, respectively, regulating the Treg/Th17 cell differentiation. In conclusions, moxibustion therapy plays a possible regulatory function in rebalancing Treg/Th17 cells by regulating miR-144-3p and its target gene Hif1a to treat CIA.