Mechanisms of S100A10 regulating the proliferation and inflammation of psoriatic keratinocytes by targeting NF-κB and STAT3
10.3760/cma.j.cn121382-20250109-00209
- VernacularTitle:S100A10通过靶向NF-κB和STAT3调控银屑病角质形成细胞增殖及炎症的机制研究
- Author:
Jing GENG
1
;
Qingqing LI
Author Information
1. 西安交通大学附属红会医院皮肤科与医疗美容科，西安 710000
- Keywords:
Psoriasis;
Keratinocytes;
Proliferation;
Inflammation;
S100 calcium binding protein A10;
Signal transduction and activator of transcription 3
- From:
International Journal of Biomedical Engineering
2025;48(2):165-173
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the mechanisms of S100 calcium binding protein A10 (S100A10) regulating the proliferation and inflammation of psoriatic keratinocytes by targeting nuclear factor-κB (NF-κB) and signal transduction and activator of transcription 3 (STAT3).Methods:Skin lesion tissues from psoriasis patients (10 cases) and normal skin tissues from healthy control (10 cases) who underwent plastic surgery at the Department of Dermatology and Medical Cosmetology, Honghui Hospital Affiliated to Xi′an Jiaotong University from January to June 2024 were collected. The pCMV plasmid and small interfering RNA plasmid were transfected into human immortalized epidermal HaCaT cells to establish the S100A10 overexpression group and the S100A10 knockdown group, respectively. Untreated HaCaT cells were used as the control group. NF-κB and STAT3 pathway inhibitors were added to the cells in the S100A10 overexpression group to create two subgroups: the S100A10 overexpression+NF-κB inhibitor group and the S100A10 overexpression+STAT3 inhibitor group. The effect of S100A10 on HaCaT cell proliferation was determined by cell counting kit-8 assay. The effect of S100A10 on HaCaT cell apoptosis was detected by extracellular phosphatidylserine binding protein V-fluorescein isothiocyanate/ propidium iodide double staining. The expression of S100A10 in normal skin tissue and skin lesion tissues of psoriasis patients was detected by Western blotting. The relative expression of apoptosis-related protein cysteine aspartic acid specific protease-3 (Caspase-3) and NF-κB and STAT3 pathway-related proteins phosphorylated NF-κB p65 (p-NF-κB p65) and phosphorylated STAT3 (p-STAT3) was detected. The effects of S100A10 on the relative expression of interleukin (IL)-6, tumor necrosis factor-α ( TNF-α), IL-1β, and CXC chemokine ligand 8 ( CXCL8) mRNA were detected by real-time reverse transcription-PCR. The effects of S100A10 on the levels of IL-6, TNF-α, IL-1β and CXCL8 in cell supernatant were detected by enzyme-linked immunosorbent assay. One-way analysis of variance and independent t test were used for data analysis. Results:The expression of S100A10 protein in the skin lesion tissues of psoriasis patients was higher than that in normal skin tissues (1.58±0.13 vs 1.00±0.09, P<0.05). Compared with the control group, the cell survival rates of the S100A10 knockdown group [(99.89±1.03)% vs (82.24±6.03)%], the relative expression of p-NF-κB p65 (0.47±0.06 vs 0.21±0.03) and p-STAT3 protein (0.59±0.12 vs 0.17±0.03), the relative expression of IL-6 (0.98±0.12 vs 0.63±0.07), TNF-α (0.97±0.13 vs 0.71±0.09), IL-1β (1.02±0.14 vs 0.51±0.09) and CXCL8 mRNA (1.01±0.16 vs 0.59±0.11), the levels of IL-6 [(27.69±3.47) pg/ml vs (13.65±2.11) pg/ml], TNF-α [(19.21±2.16) pg/ml vs (10.06±1.44) pg/ml], IL-1β [(15.52±1.03) pg/ml vs (5.17±0.96) pg/ml] and CXCL8 [(62.87±8.48) pg/ml vs (47.11±6.35) pg/ml] in the cell supernatant were lower (all P<0.05), and the cell apoptosis rate [(15.41±2.37)% vs (26.38±4.16)%] and Caspase-3 protein relative expression (0.31±0.04 vs 0.74±0.12) were higher (both P<0.05). The cell survival rates [(145.24±6.03)%], the relative expression of p-NF-κB p65 and p-STAT3 protein (2.37±0.25, 3.98±0.47), the relative expression of IL-6, TNF-α, IL-1β and CXCL8 mRNA (2.27±0.64, 3.31±0.61, 2.74±0.43, 3.11±0.48), the levels of IL-6, TNF-α, IL-1β and CXCL8 in the cell supernatant [(51.17±7.95), (33.65±4.19), (29.95±4.07), (79.97±10.32) pg/ml] in the S100A10 overexpression group were higher than those in the control group (all P<0.05), and the cell apoptosis rate [(5.09±0.73)%] and Caspase-3 protein relative expression (0.09±0.01) were lower than those in the control group (both P<0.05). The cell survival rates [(123.65±9.42)%, (122.94±8.14)%], the relative expression of p-NF-κB p65 (1.51±0.19, 1.49±0.21) and p-STAT3 protein (1.67±0.29, 1.69±0.31), the relative expression of IL-6 (1.69±0.14, 1.73±0.15), TNF-α (1.92±0.27, 1.94±0.25), IL-1β (1.85±0.16, 1.87±0.21) and CXCL8 mRNA (2.02±0.34, 2.01±0.39), the levels of IL-6 [(35.55±5.12), (36.18±5.24) pg/ml], TNF-α [(25.17±3.08), (25.23±3.17) pg/ml], IL-1β [(22.08±3.11), (22.11±3.24) pg/ml] and CXCL8 [(70.04±9.31), (70.11±10.29) pg/ml] in the cell supernatant in the S100A10 overexpression+NF-κB inhibitor group and the S100A10 overexpression+STAT3 inhibitor group were higher than those in S100A10 knockdown group (all P<0.05), and the cell apoptosis rates [(20.13±4.62)%, (20.21±4.91)%] and Caspase-3 protein relative expression (0.15±0.03, 0.16±0.04) were lower than those in S100A10 knockdown group (all P<0.05).The cell survival rates, the relative expression of p-NF-κB p65 and p-STAT3 protein, the relative expression of IL-6, TNF-α, IL-1β and CXCL8 mRNA, the levels of IL-6, TNF-α, IL-1β and CXCL8 in the cell supernatant in the S100A10 overexpression+NF-κB inhibitor group and the S100A10 overexpression+STAT3 inhibitor group were lower than those in S100A10 overexpression group (all P<0.05), and the cell apoptosis rates and Caspase-3 protein relative expression were higher than those in S100A10 overexpression group (all P<0.05). Conclusions:S100A10 may regulate the proliferation and inflammation of keratinocytes in psoriasis by targeting NF-κB and STAT3.
