实验动物科学 ›› 2024, Vol. 41 ›› Issue (6): 41-46.DOI: 10. 3969 / j. issn. 1006-6179. 2024. 06. 008

• 论著 • 上一篇    下一篇

敲除 SIRT3 缓解胶原诱导类风湿关节炎大鼠模型的分析

  

  1. ( 1. 山东省青岛市第五人民医院风湿病科,青岛 266002)  ( 2. 山东省烟台市蓬莱中医医院感染性疾病科,烟台 265600)
    ( 3. 中国人民解放军总医院第五医学中心感染病医学部研究所,北京 100039)
  • 收稿日期:2023-02-07 出版日期:2024-12-28 发布日期:2025-01-15
  • 通讯作者: 李爱民( 1969—) ,女,硕士,主任医师,研究方向为风湿免疫性疾病,E-mail:lamfsk@ 163. com。
  • 作者简介:高 萍( 1982—) ,女,硕士,主治医师,研究方向为中医药治疗风湿免疫性疾病,E-mail:xinyuan58588@ 126. com。
  • 基金资助:
    山东省青岛市中医药科技项目( 2021-zyyq07)

Elimination of SIRT3 Alleviates Collagen-induced Rheumatoid Arthritis in Rat Model

  1. ( 1. Department of Rheumatology,Qingdao Fifth People’ s Hospital, Qingdao 266002,China)
    ( 2. Department of Infectious Diseases,Penglai Hospital of Traditional Chinese Medicine,Yantai 265600,China
    ( 3. Department of Infectious Disease Medicine, the Fifth Medical Center of Chinese PLA General Hospital, Beijing 100039,China)
  • Received:2023-02-07 Online:2024-12-28 Published:2025-01-15

摘要:

目的 利用 CRISPR 技术敲除 SIRT3 胶原来诱导建立类风湿关节炎大鼠模型,分析对其免疫紊乱和软骨损伤的影响。 方法 实验分成 WT 组(野生型) 、KO 组( CRISPR 技术敲除 SIRT3) 、WT-RA 组( 野生型类风湿关节炎大鼠模型) 、KO-RA 组(CRISPR 技术敲除 SIRT3 类风湿关节炎大鼠模型) 。 检测大鼠胸腺指数和脾指数,Western blot方法检测 SIRT3、MMP-2、MMP-3、MMP-9 蛋白表达,流式细胞术分析 CD4 + / T 淋巴细胞和 CD8 + / T 淋巴细胞百分比,ELISA 法检测血清中 Th1( TNF-α、IFN-γ) 和 Th2( IL-4、IL-10) 细胞因子。 结果 与 WT 组比较,WT-RA 组 SIRT3、CD4 + 、CD4 + / CD8 + 、胸腺指数、脾指数、TNF-α、IFN-γ、MMP-2、MMP-3、MMP-9 水平显著升高,CD8 + 、IL-4、IL-10 水平显著降低(P<0. 05) 。 与 KO 组比较,KO-RA 组 CD4 + 、CD4 + / CD8 + 、胸腺指数、脾指数、TNF-α、IFN-γ、MMP-2、MMP-3、MMP-9 水平显著升高,CD8 + 、IL-4、IL-10 水平显著降低( P < 0. 05) 。 与 WT-RA 组比较,KO-RA 组 SIRT3、CD4 + 、CD4 + / CD8 + 、胸腺指数、脾指数、TNF-α、IFN-γ、MMP-2、MMP-3、MMP-9 水平显著降低,CD8 + 、IL-4、IL-10 水平显著升高(P<0. 05) 。 结论 利用 CRISPR 技术敲除 SIRT3 缓解胶原诱导的类风湿关节炎大鼠造成的免疫系统的紊乱和软骨组织的损伤。

关键词:

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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