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TAGLN2 Exacerbates Acute Pancreatitis-Induced Liver Injury by Increasing Hepatocyte Pyroptosis via Kupffer Cells-Mediated Inflammatory Response

Huigeng Zhao

Huigeng Zhao

Department of General Surgery, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
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Zhao, Huigeng
, 
Yalan Luo

Yalan Luo

Department of General Surgery, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
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Luo, Yalan
, 
Xi Chen

Xi Chen

Department of General Surgery, Affiliated Zhongshan Hospital of Dalian UniversityDalian, China
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Chen, Xi
, 
Ju Wu

Ju Wu

Department of General Surgery, Affiliated Zhongshan Hospital of Dalian UniversityDalian, China
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Wu, Ju
, 
Zizhao Zhu

Zizhao Zhu

Department of General Surgery, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
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Zhu, Zizhao
 und 
Hailong Chen

Hailong Chen

Department of General Surgery, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
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Chen, Hailong
  
05. Juni 2025
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Artikel-Kategorie: Original
Online veröffentlicht: 05. Juni 2025
Eingereicht: 04. Dez. 2024
Akzeptiert: 07. März 2025
DOI: https://doi.org/10.2478/aite-2025-0014
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© 2025 Huigeng Zhao et al., published by Sciendo
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Fig 1.

TAGLN2 is upregulated in AP-induced liver injury. Mice were treated with cerulein to induce AP. (a) The pathological changes in pancreatic and liver tissues were assessed by H&E staining. Scale bar = 200 μm. (b) The serum concentrations of amylase and lipase were measured by ELISA. (c) The serum concentrations of ALT and AST were measured by ELISA. (d) The relative mRNA expression of TAGLN2 in hepatocytes was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (e) The relative protein expression of TAGLN2 in hepatocytes was examined by western blotting. Data were expressed after being normalized with β-actin. (f) The relative mRNA expression of TAGLN2 in Kupffer cells was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (g) The relative protein expression of TAGLN2 in Kupffer cells was examined by western blotting. Data were expressed after being normalized with β-actin. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. sham. ALT, alanine aminotransferase; AP, acute pancreatitis; AST, aspartate transaminase; ELISA, enzyme-linked immunosorbent assay; H&E, Hematoxylin and eosin; TAGLN2, transgelin-2.
TAGLN2 is upregulated in AP-induced liver injury. Mice were treated with cerulein to induce AP. (a) The pathological changes in pancreatic and liver tissues were assessed by H&E staining. Scale bar = 200 μm. (b) The serum concentrations of amylase and lipase were measured by ELISA. (c) The serum concentrations of ALT and AST were measured by ELISA. (d) The relative mRNA expression of TAGLN2 in hepatocytes was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (e) The relative protein expression of TAGLN2 in hepatocytes was examined by western blotting. Data were expressed after being normalized with β-actin. (f) The relative mRNA expression of TAGLN2 in Kupffer cells was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (g) The relative protein expression of TAGLN2 in Kupffer cells was examined by western blotting. Data were expressed after being normalized with β-actin. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. sham. ALT, alanine aminotransferase; AP, acute pancreatitis; AST, aspartate transaminase; ELISA, enzyme-linked immunosorbent assay; H&E, Hematoxylin and eosin; TAGLN2, transgelin-2.

Fig 2.

TAGLN2 knockout reduces AP and AP-induced liver injury. (a) The relative mRNA expression of TAGLN2 in hepatocytes was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (b) The relative protein expression of TAGLN2 in hepatocytes was examined by western blotting. Data were expressed after being normalized with β-actin. (c) The relative mRNA expression of TAGLN2 in Kupffer cells was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (d) The relative protein expression of TAGLN2 in Kupffer cells was examined by western blotting. Data were expressed after being normalized with β-actin. (e) The pathological changes in pancreatic and liver tissues were assessed by HE staining. Scale bar = 200 μm. (f) The serum concentrations of amylase and lipase were measured by ELISA. (g) The serum concentrations of ALT and AST were measured by ELISA. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. sham + TAGLN2 WT; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. AP + TAGLN2 WT. ALT, alanine aminotransferase; AP, acute pancreatitis; AST, aspartate transaminase; ELISA, enzyme-linked immunosorbent assay; TAGLN2, transgelin-2; WT, wild type.
TAGLN2 knockout reduces AP and AP-induced liver injury. (a) The relative mRNA expression of TAGLN2 in hepatocytes was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (b) The relative protein expression of TAGLN2 in hepatocytes was examined by western blotting. Data were expressed after being normalized with β-actin. (c) The relative mRNA expression of TAGLN2 in Kupffer cells was examined by RT-qPCR. Data were expressed after being normalized with GAPDH. (d) The relative protein expression of TAGLN2 in Kupffer cells was examined by western blotting. Data were expressed after being normalized with β-actin. (e) The pathological changes in pancreatic and liver tissues were assessed by HE staining. Scale bar = 200 μm. (f) The serum concentrations of amylase and lipase were measured by ELISA. (g) The serum concentrations of ALT and AST were measured by ELISA. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. sham + TAGLN2 WT; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. AP + TAGLN2 WT. ALT, alanine aminotransferase; AP, acute pancreatitis; AST, aspartate transaminase; ELISA, enzyme-linked immunosorbent assay; TAGLN2, transgelin-2; WT, wild type.

Fig 3.

TAGLN2 knockout decreases liver tissue pyroptosis in AP mice. TAGLN2 KO mice were treated with cerulein to induce AP. (a) The relative protein expressions of NLRP3, GSDMD-N, caspase-1, IL-1β, and IL-18 in liver tissues were examined by western blotting. Data were expressed after being normalized with β-actin. (b) The concentrations of TNF-α, IL-1β, and IL-18 in liver tissues were measured by ELISA. (c) The level of F4/80 was detected by immunofluorescence assay. Scale bar = 200 μm. *p < 0.05,**p < 0.01 ***p < 0.001 vs. sham + TAGLN2 WT; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. AP + TAGLN2 WT. AP, acute pancreatitis; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; KO, knock out; TAGLN2, transgelin-2; WT, wild type.
TAGLN2 knockout decreases liver tissue pyroptosis in AP mice. TAGLN2 KO mice were treated with cerulein to induce AP. (a) The relative protein expressions of NLRP3, GSDMD-N, caspase-1, IL-1β, and IL-18 in liver tissues were examined by western blotting. Data were expressed after being normalized with β-actin. (b) The concentrations of TNF-α, IL-1β, and IL-18 in liver tissues were measured by ELISA. (c) The level of F4/80 was detected by immunofluorescence assay. Scale bar = 200 μm. *p < 0.05,**p < 0.01 ***p < 0.001 vs. sham + TAGLN2 WT; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. AP + TAGLN2 WT. AP, acute pancreatitis; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; KO, knock out; TAGLN2, transgelin-2; WT, wild type.

Fig 4.

TAGLN2 Knockdown in Kupffer cells mitigates pyroptosis of co-cultured hepatocytes. Kupffer cells were isolated from WT mice. Then, Kupffer cells were transfected with shTAGLN2 to downregulate the expression of TAGLN2, and treated with LPS. (a) The relative protein expression of TAGLN2 in Kupffer cells was examined by western blotting. Data were expressed after being normalized with β-actin. (b) The concentrations of TNF-α, IL-1β, and IL-18 in the supernatant of Kupffer cells were measured by ELISA. (c) The supernatant of Kupffer cells was yielded as the conditional medium, and was used to incubate with hepatocytes isolated from WT mice. The relative protein expressions of NLRP3, GSDMD-N, caspase-1, IL-1β, and IL-18 in liver tissues were examined by western blotting. Data were expressed after being normalized with β-actin. (d) The pyroptosis rate of hepatocytes was determined by flow cytometry after hepatocytes isolated from WT mice were incubated with the supernatant of Kupffer cells. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. shNC; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. shTAGLN2. ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; TAGLN2, transgelin-2; WT, wild type.
TAGLN2 Knockdown in Kupffer cells mitigates pyroptosis of co-cultured hepatocytes. Kupffer cells were isolated from WT mice. Then, Kupffer cells were transfected with shTAGLN2 to downregulate the expression of TAGLN2, and treated with LPS. (a) The relative protein expression of TAGLN2 in Kupffer cells was examined by western blotting. Data were expressed after being normalized with β-actin. (b) The concentrations of TNF-α, IL-1β, and IL-18 in the supernatant of Kupffer cells were measured by ELISA. (c) The supernatant of Kupffer cells was yielded as the conditional medium, and was used to incubate with hepatocytes isolated from WT mice. The relative protein expressions of NLRP3, GSDMD-N, caspase-1, IL-1β, and IL-18 in liver tissues were examined by western blotting. Data were expressed after being normalized with β-actin. (d) The pyroptosis rate of hepatocytes was determined by flow cytometry after hepatocytes isolated from WT mice were incubated with the supernatant of Kupffer cells. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. shNC; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. shTAGLN2. ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; TAGLN2, transgelin-2; WT, wild type.

Fig 5.

TAGLN2 enhances the activation of ANXA2/NF-κB axis in Kupffer cells. (a) The relative protein expression of ANXA2, p-P65 and P65 in Kupffer cells from mice was examined by western blotting. Data were expressed after being normalized with β-actin. ***p < 0.001 vs. sham + TAGLN2 WT; #p < 0.05 and ##p < 0.01 vs. AP + TAGLN2 WT. (b) The relative protein expression of ANXA2, p-P65 and P65 in Kupffer cells in vitro was examined by western blotting. Data were expressed after being normalized with β-actin. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. shNC; #p < 0.05,##p < 0.01 and ###p < 0.001 vs. shTAGLN2. AP, acute pancreatitis; LPS, lipopolysaccharide; TAGLN2, transgelin-2; WT, wild type.
TAGLN2 enhances the activation of ANXA2/NF-κB axis in Kupffer cells. (a) The relative protein expression of ANXA2, p-P65 and P65 in Kupffer cells from mice was examined by western blotting. Data were expressed after being normalized with β-actin. ***p < 0.001 vs. sham + TAGLN2 WT; #p < 0.05 and ##p < 0.01 vs. AP + TAGLN2 WT. (b) The relative protein expression of ANXA2, p-P65 and P65 in Kupffer cells in vitro was examined by western blotting. Data were expressed after being normalized with β-actin. *p < 0.05, **p < 0.01 and ***p < 0.001 vs. shNC; #p < 0.05,##p < 0.01 and ###p < 0.001 vs. shTAGLN2. AP, acute pancreatitis; LPS, lipopolysaccharide; TAGLN2, transgelin-2; WT, wild type.
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