Gelsolin impairs barrier function in pancreatic ductal epithelial cells by actin filament depolymerization in hypertriglyceridemia‑induced pancreatitis in vitro

Figures

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  Figure 1. Effect of intracellular calcium levels on GSN expression in HPDE6-C7 cells treated with CAE and TG. (A) Changes in intracellular calcium levels based on fluorescence imaging (x200 magnification) and (B) semi-quantitative analysis. (C) Protein expression levels of GSN, as determined by western blotting and (D) semi-quantitative analysis. (E) Relative mRNA expression levels of GSN, as determined by reverse transcription-quantitative polymerase chain reaction. The experiments were repeated at least three times. Data are presented as the mean ± standard deviation. **P<0.01. BC, blank control; CAE, caerulein; GSN, gelsolin; KD, knockdown; NC, negative control; TG, triglycerides.

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  Figure 2. Effect of GSN on the dynamics of actin filaments in HPDE6-C7 cells treated with CAE and TG. The changes in actin filaments induced by silencing GSN were analyzed by tetramethyl rhodamine isothiocyanate-phalloidin immunofluorescence under an upright fluorescence microscope (x1,000 magnification, white arrows indicate actin filaments). The experiment was repeated at least three times. BC, blank control; CAE, caerulein; GSN, gelsolin; KD, knockdown; NC, negative control; TG, triglycerides.

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  Figure 3. Effect of gelsolin on the major components of cell-cell junctions in HPDE6-C7 cells treated with CAE and TG. Relative mRNA expression levels of (A) E-cadherin, (B) nectin-2, (C) ZO-1 and (D) occludin, as determined by reverse transcription-quantitative polymerase chain reaction. (E) Western blot analysis and semi-quantification of the protein expression levels of (F) E-cadherin, (G) nectin-2, (H) ZO-1 and (I) occludin. The experiments were repeated at least three times. Data are presented as the mean ± standard deviation. *P<0.05, **P<0.01. BC, blank control; CAE, caerulein; KD, knockdown; NC, negative control; TG, triglycerides.

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  Figure 4. Effect of gelsolin on TJ ultrastructure and the permeability of HPDE6-C7 cells treated with CAE and TG. (A) Ultrastructure of TJs, as determined by transmission electron microscopy (x40,000 magnification); red arrows indicate TJs. (B) Cell permeability was analyzed by FITC-dextran fluorescence. The experiments were repeated at least three times. Data are presented as the mean ± standard deviation. **P<0.01. BC, blank control; CAE, caerulein; KD, knockdown; NC, negative control; TG, triglycerides; TJ, tight junction.