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CLEC5A Knockdown Reduces Oxidative Stress and Inflammation Caused by Lipopolysaccharide in Renal Tubular Epithelial Cells via the NF-κB/NLRP3 Signaling Pathway Cover

CLEC5A Knockdown Reduces Oxidative Stress and Inflammation Caused by Lipopolysaccharide in Renal Tubular Epithelial Cells via the NF-κB/NLRP3 Signaling Pathway

Archivum Immunologiae et Therapiae Experimentalis
Volume 74 (2026): Issue 1 (January 2026)
By: Zunjiang Chen,  Chengfeng Pan,  Xuexiu Chen,  Shangli Xie,  Yuechuan Lin and  Xingken Fan  
Open Access
|Oct 2025

Figures & Tables

Fig 1.

CLEC5A knockdown protects HK-2 cells from LPS-induced injury. (A, B) Detection of CLEC5A expression in HK-2 cells by Western blotting. (C) CCK-8 assay for cell viability. (D) Measure LDH production using a commercially available LDH release assay kit. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CCK-8, cell counting kit-8; CLEC5A, C-type lectin domain family 5 member A; GAPDH, LDH, lactate dehydrogenase; LPS, lipopolysaccharide; SD, standard deviation; shNC, shcontrol.
CLEC5A knockdown protects HK-2 cells from LPS-induced injury. (A, B) Detection of CLEC5A expression in HK-2 cells by Western blotting. (C) CCK-8 assay for cell viability. (D) Measure LDH production using a commercially available LDH release assay kit. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CCK-8, cell counting kit-8; CLEC5A, C-type lectin domain family 5 member A; GAPDH, LDH, lactate dehydrogenase; LPS, lipopolysaccharide; SD, standard deviation; shNC, shcontrol.

Fig 2.

CLEC5A knockdown improves HK-2 cells from LPS-induced inflammation. ELISA detects TNF-α (A), IL-6 (B), IL-1β, and (C) levels in HK-2 cells culture fluid. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CLEC5A, C-type lectin domain family 5 member A; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; SD, shNC, TNF-α, tumor necrosis factor-α.
CLEC5A knockdown improves HK-2 cells from LPS-induced inflammation. ELISA detects TNF-α (A), IL-6 (B), IL-1β, and (C) levels in HK-2 cells culture fluid. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CLEC5A, C-type lectin domain family 5 member A; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; SD, shNC, TNF-α, tumor necrosis factor-α.

Fig 3.

CLEC5A knockdown improves HK-2 cells from LPS-induced oxidative stress. MDA (A), SOD (B), and GSH-Px (C) levels were measured using commercially available kits in HK-2 cells. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CLEC5A, C-type lectin domain family 5 member A; GSH-Px, glutathione peroxidase; LPS, lipopolysaccharide; MDA, malondialdehyde; SD, shNC, SOD, superoxide dismutase.
CLEC5A knockdown improves HK-2 cells from LPS-induced oxidative stress. MDA (A), SOD (B), and GSH-Px (C) levels were measured using commercially available kits in HK-2 cells. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CLEC5A, C-type lectin domain family 5 member A; GSH-Px, glutathione peroxidase; LPS, lipopolysaccharide; MDA, malondialdehyde; SD, shNC, SOD, superoxide dismutase.

Fig 4.

CLEC5A knockdown can block the activation of NF-κB/NLRP3 signaling pathway. Western blotting to detect p65, p-p65, NLRP3, ASC protein expression. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. ASC, CLEC5A, C-type lectin domain family 5 member A; LPS, lipopolysaccharide; NLRP3, NLR Family, Pyrin Domain Containing Protein 3; SD, shNC.
CLEC5A knockdown can block the activation of NF-κB/NLRP3 signaling pathway. Western blotting to detect p65, p-p65, NLRP3, ASC protein expression. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. ASC, CLEC5A, C-type lectin domain family 5 member A; LPS, lipopolysaccharide; NLRP3, NLR Family, Pyrin Domain Containing Protein 3; SD, shNC.

Fig 5.

Activation of NF-κB reverses the effects of CLEC5A knockdown on LPS-treated HK-2 cells. (A) Western blotting to detect p65, p-p65, NLRP3, and ASC protein expression. (B) CCK-8 assay for cell viability. (C) Measure LDH production using a commercially available LDH release assay kit. (D) ELISA detects IL-1β, IL-6, and TNF-α levels. (E) MDA, SOD, and GSH-Px levels were measured using commercially available kits in HK-2 cells. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. ASC, CCK-8, cell counting kit-8; CLEC5A, C-type lectin domain family 5 member A; ELISA, enzyme-linked immunosorbent assay; GSH-Px, glutathione peroxidase; IL, interleukin; LDH, lactate dehydrogenase; LPS, lipopolysaccharide; MDA, malondialdehyde; NF-κB, NLRP3, NLR Family, Pyrin Domain Containing Protein 3; SD, shNC, SOD, superoxide dismutase; TNF-α, tumor necrosis factor-α.
Activation of NF-κB reverses the effects of CLEC5A knockdown on LPS-treated HK-2 cells. (A) Western blotting to detect p65, p-p65, NLRP3, and ASC protein expression. (B) CCK-8 assay for cell viability. (C) Measure LDH production using a commercially available LDH release assay kit. (D) ELISA detects IL-1β, IL-6, and TNF-α levels. (E) MDA, SOD, and GSH-Px levels were measured using commercially available kits in HK-2 cells. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. ASC, CCK-8, cell counting kit-8; CLEC5A, C-type lectin domain family 5 member A; ELISA, enzyme-linked immunosorbent assay; GSH-Px, glutathione peroxidase; IL, interleukin; LDH, lactate dehydrogenase; LPS, lipopolysaccharide; MDA, malondialdehyde; NF-κB, NLRP3, NLR Family, Pyrin Domain Containing Protein 3; SD, shNC, SOD, superoxide dismutase; TNF-α, tumor necrosis factor-α.
DOI: https://doi.org/10.2478/aite-2026-0001 | Journal eISSN: 1661-4917
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Language: English
Submitted on: May 22, 2025
Accepted on: Aug 13, 2025
Published on: Oct 31, 2025
Published by: Hirszfeld Institute of Immunology and Experimental Therapy
In partnership with: Paradigm Publishing Services
Publication frequency: 1 issue per year
Keywords:
S-AKI,
CLEC5A,
NF-κB/NLRP3,
Inflammation,
Oxidative stress,
Renal tubular epithelial cell
Related subjects:
Medicine,
Basic medical science,
Biochemistry,
Immunology,
Clinical medicine,
Clinical medicine, other,
Clinical chemistry

© 2025 Zunjiang Chen, Chengfeng Pan, Xuexiu Chen, Shangli Xie, Yuechuan Lin, Xingken Fan, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Volume 74 (2026): Issue 1 (January 2026)