Effect of low expressed SIRT-6 gene on inflammatory reaction and autophagy in monocytes
10.3760/cma.j.cn141217-20231130-00158
- VernacularTitle:沉默信息调节因子6基因低表达对单核细胞炎症反应 细胞自噬的调控作用及对痛风炎症的影响
- Author:
Jing LI
1
;
Jiangping HE
;
Juan XU
;
Tianxue ZHAO
;
Siyue LIU
;
Haiyan QIU
;
Yuhong ZHAN
Author Information
1. 杭州市老年病医院内分泌科，杭州 310022
- Keywords:
Gout;
Silent information regulator;
Monocytes;
Inflammatory reaction;
Autophagy;
Urate;
Rapamycin
- From:
Chinese Journal of Rheumatology
2024;28(8):558-565
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of low expression of silencing information regulator-6 (SIRT-6) on inflammatory reaction and autophagy in monocytes.Methods:Human acute monocytic leukemia cell line THP-1 was transfected with si-SIRT6 to establish THP-1 cell line with low expressed SIRT-6. The cells were divided into control group, MUS group and MUS+ RAPA group. Cells in control group were cultured with medium added with PBS, cells in MUS Group were cultured with medium added with MUS, and cells in MUS+ RAPA Group were added with MUS and Rapamycin. Cells in each group were cultured for 48 hours. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the supernatant of each group were detected by enzyme-linked immunosorbent assay (ELISA). The gene expression levels of autophagy-associated protein-5 (ATG-5), Beclin-1, lysosomal-associated membrane protein-1 (LAMP-1), microtubule-associated protein-1 light chain 3B (LC3B) and p62 in cells of each group were detected by Q-PCR. The protein expression levels of p62, ATG-5 and LC3B Ⅱ/LC3BⅠ in cells of each group. The one-way analysis of variance (ANOVA) was used for the measurement data in multi-groups, and the LSD- t test was used for the measurement data in both groups. Results:The gene and protein expression of SIRT-6 in THP-1 cells decreased significantly after si-SIRT6 transfection (Gene: 1.09±0.08 vs. 0.57±0.03, t=14.91, P<0.001; Protein: 0.21±0.04 vs. 0.12±0.03, t=4.41, P=0.070). The levels of IL-1β, IL-6, and TNF-α in the supernatant of si-SIRT6/si-SIRT6 NC-transfected THP-1 cells increased significantly by MUS ( P<0.05), and the levels of IL-1β, IL-6, and TNF-α in the supernatant of cells further increased by MUS ( P<0.05). The levels of IL-1β, IL-6 and TNF-α in the supernatant of si-SIRT6-transfected THP-1 cells increased significantly compared with those of si-SIRT6 NC-transfected THP-1 cells ( P<0.05). The gene expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly decreased by MUS ( P<0.05), the gene expression of ATG-5, Beclin-1, LAMP-1 and LC3B in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly increased by MUS ( P<0.05). The gene expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further decreased by RAPA ( P<0.05), the gene expression of ATG-5, Beclin-1, LAMP-1 and LC3B in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further increased by RAPA ( P<0.05). The gene expression level of p62 in si-SIRT6 transfected THP-1 cells significantly decreased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05), and the gene expression level of ATG-5, LC3B, Beclin-1 and LAMP-1 significantly increased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05). The protein expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly decreased by MUS ( P<0.05), the protein expression of ATG-5 and LC3B Ⅱ/LC3BⅠ protein in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly increased by MUS ( P<0.05). The protein expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further decreased by RAPA P<0.05), the protein expression of ATG-5 and LC3B Ⅱ/LC3BⅠ in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further increased by RAPA ( P<0.05). The protein expression level of p62 in si-SIRT6 transfected THP-1 cells significantly decreased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05), and the protein expression level of ATG-5 and LC3B Ⅱ/LC3BⅠ significantly increased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05). Conclusion:Low expression of SIRT-6 gene can promote inflammatory reaction and autophagy in monocytes, and Monosodium urate and autophagy agonist rapamycin can aggravate inflammatory reaction and autophagy.
