Abstract: Objective The rat model of rheumatoid arthritis was established by using CRISPR technology to knockout SIRT3, and the effects on immune disorder and cartilage damage were analyzed. Method The experiment was divided into WT group ( wild type) , KO group ( SIRT3 was knockout by CRISPR technology) , WT-RA group (wild type rat model of rheumatoid arthritis) , KO-RA group ( a rat model of rheumatoid arthritis in which SIRT3 was knocked out by CRISPR) . The thymus index and spleen of rats were detected. Protein expression of SIRT3, MMP-2, MMP-3, and MMP-9 were detected by Western blot, and the percentage of CD4 + / T lymphocytes and CD8 + / T lymphocytes was analyzed by flow cytometry. The serum cytokines of Th1 ( TNF-α, IFN-γ) and Th2 ( IL-4, IL-10 ) were detected by ELISA. Result Compared with WT group, the levels of SIRT3, CD4 + , CD4 + / CD8 + , thymus index, spleen index, TNF-α, IFN-γ, MMP-2, MMP-3 and MMP-9 in WT-RA group were significantly increased, while the levels of CD8 + , IL-4 and IL-10 were significantly decreased (P<0. 05) . Compared with KO group, the levels of CD4 + , CD4 + / CD8 + , thymus index, spleen index, TNF-α, IFN-γ, MMP- 2, MMP-3 and MMP-9 in KO-RA group were significantly increased, while the levels of CD8 + , IL-4 and IL-10 were significantly decreased ( P < 0. 05 ) . Compared with WT-RA group, the levels of SIRT3, CD4 + , CD4 + / CD8 + , thymus index, spleen index, TNF-α, IFN-γ, MMP-2, MMP-3 and MMP-9 in KO- RA group were significantly decreased, while the levels of CD8 + , IL-4 and IL-10 were significantly increased (P<0. 05) . Conclusion Using CRISPR technology to knock out SIRT3 alleviated the immune disorder and cartilage damage in rheumatoid arthritis rats induced by collagen.

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