Journal: International Journal of Molecular Sciences

Article Title: Esaxerenone Attenuates Aldosterone-Induced Renal Fibrosis by Suppressing Fibroblast-to-Lymphatic Endothelial-like Cell Transdifferentiation

doi: 10.3390/ijms27104297

Figure Lengend Snippet: Esaxerenone significantly reduces renal fibrosis in aldosterone-infused rats. ( A ) Masson staining, Sirius red staining, and Vimentin/COL-III immunohistochemistry were performed to detect pathological changes and fibrosis in rat kidneys ( n = 6); scale bars, 100 (Masson) and 50 (others). All in µm. ( B ) Western blot was used to determine the protein expression levels of α-SMA and Vimentin in rat kidneys ( n = 6). ( C ) Western blot was employed to measure the protein expression levels of α-SMA and Vimentin in RKFs after aldosterone stimulation ( n = 6). ( D ) Serum creatinine and blood urea nitrogen were detected for renal function evaluation ( n = 3). All data are presented as mean ± SD, and one-way ANOVA was used to examine intergroup differences (* p < 0.05 vs. Sham group; # p < 0.05 vs. ALD group).

Article Snippet: The RKFs were divided into six groups: the CON group, the ALD group (stimulated with 10 −7 mol/L aldosterone for 24 h), the ALD + ESA group (pretreated with 10 −6 mol/L esaxerenone 2 h prior to aldosterone treatment), VEGFC group (MCE, Shanghai, China; Cat#: HY- P74474 , 100 ng/mL), ALD+ VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-P7025, 20 ng/mL; stimulation for 48 h), and VEGFC + VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-132305, 20 ng/mL) and stimulated for 24 h.

Techniques: Staining, Immunohistochemistry, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Esaxerenone Attenuates Aldosterone-Induced Renal Fibrosis by Suppressing Fibroblast-to-Lymphatic Endothelial-like Cell Transdifferentiation

doi: 10.3390/ijms27104297

Figure Lengend Snippet: Esaxerenone attenuated the transdifferentiation of renal fibroblasts into lymphatic endothelial-like cells in aldosterone-infused rats. ( A ) Immunohistochemistry staining using antibodies against LYVE-1, PDPN, VEGFR-3 and FSP-1 to examine transdifferentiation of renal fibroblasts into lymphatic endothelial-like cells in the rat kidneys ( n = 6); scale bars, 50 μm. The correlation of FSP-1 expression with the positive expressions of LYVE-1, PDPN, and VEGFR-3 was evaluated. ( B ) Western blot was employed to examine the protein levels of LYVE-1, PDPN, VEGFR-3, and FSP-1 in the rat kidneys, followed by quantitative analysis ( n = 6). ( C ) Western blot was performed to determine the protein expressions of LYVE-1 and VEGFR-3 in RKFs after aldosterone stimulation, with subsequent quantitative analysis ( n = 6). ( D ) The expressions of FSP-1 and VEGFR-3 in the kidney were analyzed by flow cytometry. First, all fibroblasts in the rat kidney were identified, and Q1 represents the proportion of fibroblasts that transdifferentiated into lymphatic endothelial-like cells among the non-immune cells ( n = 3). All data are presented as mean ± SD, and one-way ANOVA was used to examine intergroup differences (* p < 0.05 vs. Sham group; # p < 0.05 vs. ALD group).

Article Snippet: The RKFs were divided into six groups: the CON group, the ALD group (stimulated with 10 −7 mol/L aldosterone for 24 h), the ALD + ESA group (pretreated with 10 −6 mol/L esaxerenone 2 h prior to aldosterone treatment), VEGFC group (MCE, Shanghai, China; Cat#: HY- P74474 , 100 ng/mL), ALD+ VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-P7025, 20 ng/mL; stimulation for 48 h), and VEGFC + VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-132305, 20 ng/mL) and stimulated for 24 h.

Techniques: Immunohistochemistry, Staining, Expressing, Western Blot, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Esaxerenone Attenuates Aldosterone-Induced Renal Fibrosis by Suppressing Fibroblast-to-Lymphatic Endothelial-like Cell Transdifferentiation

doi: 10.3390/ijms27104297

Figure Lengend Snippet: MR activation and inflammatory factors changes in aldosterone-infused rats. ( A ) Immunohistochemistry was used to detect the expression of NR3C2, IL-1β, MCP-1, TNF-α, and VEGFC in rat kidneys ( n = 6); scale bars, 50 μm. ( B ) Western blot was performed to determine the protein expression levels of NR3C2, IL-1β, MCP-1, TNF-α, and VEGFC in rat kidneys ( n = 6); ( C ) Western blot was used to detect the protein expression of NR3C2 and VEGFC in RKFs after aldosterone stimulation ( n = 6); ( D ) Serum aldosterone detection ( n = 3). All data are presented as mean ± SD, and one-way ANOVA was used to examine intergroup differences (* p < 0.05 vs. Sham group; # p < 0.05 vs. ALD group).

Article Snippet: The RKFs were divided into six groups: the CON group, the ALD group (stimulated with 10 −7 mol/L aldosterone for 24 h), the ALD + ESA group (pretreated with 10 −6 mol/L esaxerenone 2 h prior to aldosterone treatment), VEGFC group (MCE, Shanghai, China; Cat#: HY- P74474 , 100 ng/mL), ALD+ VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-P7025, 20 ng/mL; stimulation for 48 h), and VEGFC + VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-132305, 20 ng/mL) and stimulated for 24 h.

Techniques: Activation Assay, Immunohistochemistry, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Esaxerenone Attenuates Aldosterone-Induced Renal Fibrosis by Suppressing Fibroblast-to-Lymphatic Endothelial-like Cell Transdifferentiation

doi: 10.3390/ijms27104297

Figure Lengend Snippet: Effects of esaxerenone on RKF migration and tube formation in vitro. ( A ) RKF migration assay ( n = 3); scale bars, 100 μm. ( B ) RKFs possess a certain migratory capacity in vitro and are able to form three-dimensional tubular structures. The tubular formation was observed and photographed at 0 h, 2 h and 4 h, and qualitative analysis of the network tube length was performed ( n = 3); scale bars, 100 μm. All data are presented as mean ± SD, and one-way ANOVA was used to examine intergroup differences (* p < 0.05 vs. Sham group; # p < 0.05 vs. ALD group).

Article Snippet: The RKFs were divided into six groups: the CON group, the ALD group (stimulated with 10 −7 mol/L aldosterone for 24 h), the ALD + ESA group (pretreated with 10 −6 mol/L esaxerenone 2 h prior to aldosterone treatment), VEGFC group (MCE, Shanghai, China; Cat#: HY- P74474 , 100 ng/mL), ALD+ VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-P7025, 20 ng/mL; stimulation for 48 h), and VEGFC + VEGFR-3-IN-1 group (MCE, Shanghai, China; Cat#: HY-132305, 20 ng/mL) and stimulated for 24 h.

Techniques: Migration, In Vitro