Journal: Experimental and Therapeutic Medicine

Article Title: A high salt diet impairs the bladder epithelial barrier and activates the NLRP3 and NF‑κB signaling pathways to induce an overactive bladder in vivo

doi: 10.3892/etm.2024.12651

Figure Lengend Snippet: Primer sequences for reverse transcription-quantitative PCR.

Article Snippet: The samples were then incubated at 4˚C overnight with the following primary antibodies: Rabbit anti-TJP-1 antibody (1:250; cat. no. ab276131; Abcam), rabbit anti-CLAUDIN-1 antibody (1:200; cat. no. YT0942; ImmunoWay Biotechnology Company), rabbit anti-TRPV4 antibody (1:200; cat. no. YT5833; ImmunoWay Biotechnology Company), rabbit anti-CASP1 antibody (1:200; cat. no. YP0749; ImmunoWay Biotechnology Company) and rabbit anti-NF-κB antibody (1:200; cat. no. YM8001; ImmunoWay Biotechnology Company).

Techniques: Sequencing

Journal: Experimental and Therapeutic Medicine

Article Title: A high salt diet impairs the bladder epithelial barrier and activates the NLRP3 and NF‑κB signaling pathways to induce an overactive bladder in vivo

doi: 10.3892/etm.2024.12651

Figure Lengend Snippet: A HSD in vivo impaired barrier function of bladder. (A) H&E staining and histological score of bladder tissues (scale bar, 1 mm). Data were presented as the median with interquartile range. (B) Bladder weight/body weight ratio. (C) Thickness of lamina propria and mucosal layer of the bladder. Relative mRNA expression levels of (D) inflammatory response markers, IL-1β and TNF-α, (E) tight junction proteins, TJP-1 and Claudin-1 and (F) TRPV4. (G) Representative images of histological staining and quantification of protein expression of TRPV4, TJP-1 and CLAUDIN-1 in bladder tissues sections from CON and HSD mice. Scale bar, 250 µm (n=3). (H) Correlation analysis between the mRNA expression levels of tight junction proteins and inflammation factors in the bladder and urination characteristics in CON and HSD mice. Data are presented as mean ± SD (n=6). P-values were calculated using a two-tailed unpaired Student's t test; *** P<0.001, ** P<0.01 and * P<0.05. HSD, high salt diet; CON, control; TJP-1, tight junction protein 1; TRPV4, transient receptor potential vanilloid 4; ns, non-significant; A.U., arbitrary units.

Article Snippet: The samples were then incubated at 4˚C overnight with the following primary antibodies: Rabbit anti-TJP-1 antibody (1:250; cat. no. ab276131; Abcam), rabbit anti-CLAUDIN-1 antibody (1:200; cat. no. YT0942; ImmunoWay Biotechnology Company), rabbit anti-TRPV4 antibody (1:200; cat. no. YT5833; ImmunoWay Biotechnology Company), rabbit anti-CASP1 antibody (1:200; cat. no. YP0749; ImmunoWay Biotechnology Company) and rabbit anti-NF-κB antibody (1:200; cat. no. YM8001; ImmunoWay Biotechnology Company).

Techniques: In Vivo, Staining, Expressing, Two Tailed Test, Control

Journal: Experimental and Therapeutic Medicine

Article Title: A high salt diet impairs the bladder epithelial barrier and activates the NLRP3 and NF‑κB signaling pathways to induce an overactive bladder in vivo

doi: 10.3892/etm.2024.12651

Figure Lengend Snippet: A HSD increased uroepithelial oxidative stress in SV-HUC-1 cells. Relative mRNA expression levels of (A) TJP-1 and CLAUDIN-1, (B) TRPV4 and (C) IL-1β and TNF-α in HSD-treated and CON cells. (D) Relative MPO expression levels in CON and HSD groups. (E) Representative images and quantification of intracellular ROS levels (scale bar, 200 µm). (F) Relative mRNA expression levels of NCF-1 and CYBA. (G) Relative MDA expression levels in CON and HSD groups. Data are presented as mean ± SD (n=6). P-values were calculated using a two-tailed unpaired Student's t-test; *** P<0.001 and ** P<0.01. HSD, high salt diet; CON, control; SV-HUC-1, SV40 virus transformed human uroepithelium cells; TJP-1, tight junction protein 1; TRPV4, transient receptor potential vanilloid 4; MPO, myeloperoxidase; ROS, reactive oxygen species; NCF-1, neutrophil cytosolic factor 1; CYBA, cytochrome B-245 alpha chain; MDA, malondialdehyde.

Article Snippet: The samples were then incubated at 4˚C overnight with the following primary antibodies: Rabbit anti-TJP-1 antibody (1:250; cat. no. ab276131; Abcam), rabbit anti-CLAUDIN-1 antibody (1:200; cat. no. YT0942; ImmunoWay Biotechnology Company), rabbit anti-TRPV4 antibody (1:200; cat. no. YT5833; ImmunoWay Biotechnology Company), rabbit anti-CASP1 antibody (1:200; cat. no. YP0749; ImmunoWay Biotechnology Company) and rabbit anti-NF-κB antibody (1:200; cat. no. YM8001; ImmunoWay Biotechnology Company).

Techniques: Expressing, Two Tailed Test, Control, Virus, Transformation Assay

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Quantification of TRPV4 expression. (a) Sites where tissue samples were taken and the quantification method for stained areas. Tissue was collected from 57 patients who underwent gastric resection. The following areas were sectioned in all layers: 1. greater curvature of the fornix; 2. greater curvature of the gastric body; 3. greater curvature of the antrum; 4. anterior wall of the gastric body; 5, posterior wall of the gastric body. (b) TRPV4 immunostaining in gastric tissue. (c) The images were captured using a microscope with BZ-X710 (Keyence, Osaka, Japan), and staining was quantified with hybrid cell count BZ-H4C.

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Expressing, Staining, Immunostaining, Microscopy, Cell Counting

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Comparison of stained areas between nonobese and obese cases. (a) Renal tissues were used as a positive control for TRPV4-immunostaining. (b) For the negative control, tissues were subjected to immunostaining without the primary antibody. (c) Immunostaining of the nonobese and obese stomach. (d) Comparison of stained areas in tissue samples collected from patients with and without obesity. Lines within the boxes represent median values; upper and lower lines of the boxes represent 25th and 75th percentiles, respectively; upper and lower bars outside the boxes represent the maximum and minimum, respectively ( ∗∗ p < 0.01).

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Comparison, Staining, Positive Control, Immunostaining, Negative Control

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: TRPV4 immunostaining in the stomach of patients with and without obesity.

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Immunostaining

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Fluorescence immunostaining of TRPV4 in MGN3-1 cells. TRPV4 protein expression was observed in MGN3-1 cells. Magnification: ×600. TRPV4-mediated change in [Ca 2+ ] i in MGN3-1 cells.

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Fluorescence, Immunostaining, Expressing

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: TRPV4-mediated changes in cytosolic Ca 2+ in MGN3-1 cells. The effects of GSK1016790 A and HC 067047 on [Ca 2+ ] i in MGN3-1 cells. Compared to the control level at 180 s, [Ca2+] i increased in MGN3-1 cells treated with GSK1016790 A ( n = 20, p < 0.01) but decreased in cells treated with GSK1016790 A and HC067047 ( n = 20, p < 0.01) (GSK1016790 A, TRPV4 agonist; HC067047, TRPV4 antagonist).

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Control

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Regulation of ghrelin secretion by the TRPV4 agonist and antagonist in MGN3-1 cells. (a) The amount of secreted acyl ghrelin after four-hour treatment of MGN3-1 cells with GSK1016790 A (3 μ M) or HC067047 (20 μ M) + GSK1016790 A (3 μ M). (b) The amount of secreted des-acyl ghrelin after four-hour treatment of MGN3-1 cells with GSK1016790 A (3 μ M) or HC067047 (20 μ M) + GSK1016790 A (3 μ M) ( ∗ p < 0.05; ∗∗ p < 0.01; n = 6; error bars: standard error of the mean).

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: