Role of TLR2/NF-κB signaling pathway in aluminum-induced inflammatory response in GMI-R1 cells
10.20001/j.issn.2095-2619.20231001
- VernacularTitle:TLR2/NF-κB信号通路在铝致GMI-R1细胞炎症反应中的作用
- Author:
Xiangming LIU
1
;
Shuhui ZHANG
;
Xiaoyan LI
;
Qian HU
;
Qiao NIU
Author Information
1. Department of Labor Health, Shanxi Key Laboratory of Environmental Health Damage, MOE Key Laboratory of Coal Environmental Pathogenicity and Prevention, Shanxi Medical University, Taiyuan, Shanxi 030001, China
- Publication Type:Journal Article
- Keywords:
Aluminum;
GMI-R1 cells;
Toll-like receptor 2;
Nuclear factor-κB;
Signaling pathway;
Inflammation
- From:
China Occupational Medicine
2023;50(5):481-488
- CountryChina
- Language:Chinese
-
Abstract:
{L-End}Objective To investigate the role and mechanism of Toll-like receptor 2 (TLR2)/nuclear factor-κB(NF-κB) signaling pathway in the inflammatory response induced by aluminum in the rat GMI-R1 microglia cells. {L-End}Methods GMI-R1 cells in logarithmic growth phase were randomly divided into the control group, positive control group, and low-, medium-, and high-dose groups. The cells in the three dose groups were stimulated with maltolol aluminum at concentrations of 100, 200, and 400 μmol/L, respectively. The cells in the positive control group were stimulated with lipopolysaccharide at a mass concentration of 20 mg/L, while the cells in the control group were not treated. The morphological changes of cells were observed, and the cell survival rate was evaluated by CCK-8 method after 24 hours of culture. The secretion levels of tumor necrosis factor-α (TNF-α), interleukin (IL) -12 and IL-4 were detected by enzyme-linked immunosorbent assay. The relative protein expression of TLR2, NF-κB P65, cluster of differentiation (CD) 68 and CD206 of cells was detected by Western blotting, and the expression of CD68 and CD206 of cells was detected by immunofluorescence method. {L-End}Results The results of cell morphology showed that the number of GMI-R1 cells decreased, the number of activated cells increased, the degree of cell cytoplasm filling decreased, and the cell protrusions elongated with the increase of exposure dose, showing a dose-response relationship. The cell viability of the positive control group and the medium- and high-dose groups were significantly lower than those of the control group (all P<0.05). The secretion levels of TNF-α, IL-12 and the relative expression of TLR2 and CD68 proteins increased (all P<0.05) while the secretion level of IL-4 decreased (all P<0.05) in the cells of positive control group compared with the control group. The secretion levels of TNF-α and IL-12 increased (all P<0.05) while the secretion levels of IL-4 decreased in the cells of the three doses groups (all P<0.05), compared with the control group, and all showed a dose-effect relationship. The relative expression of TLR2 protein in the cells of the three doses groups increased (all P<0.05) compared with the control group. The relative expression of NF-κB p65 and CD68 protein in the cells of the medium- and high-dose groups increased (all P<0.05), but the relative expression of CD206 protein decreased (all P<0.05) compared with the control group. The relative expression of TLR2 and NF-κB p65 protein increased (all P<0.05) while the relative expression of CD206 protein decreased (all P<0.05) in cells of the high-dose group, compared with the low- and medium-dose groups. The average fluorescence intensity of CD68 increased (all P<0.05) while the average fluorescence intensity of CD206 decreased in the cells of high-dose group and the positive control group (all P<0.05), compared with the control group. {L-End}Conclusion Aluminum participated in and promoted the inflammatory response of GMI-R1 cells through the TLR2/NF-κB signaling pathway.
