Journal: International Journal of Molecular Sciences

Article Title: Sirtuin 3 Activation by Honokiol Decreases Unilateral Ureteral Obstruction-Induced Renal Inflammation and Fibrosis via Regulation of Mitochondrial Dynamics and the Renal NF-κB-TGF-β1/Smad Signaling Pathway

doi: 10.3390/ijms21020402

Figure Lengend Snippet: Effect of honokiol on unilateral ureteral obstruction (UUO)-induced myofibroblast activation and extracellular matrix deposition. ( A ) Representative sections of kidneys from sham- and UUO-operated mice treated with Vehicle (Veh) or honokiol (HKL) were immunofluorescence-stained with α-smooth muscle actin (α-SMA) (red). The nucleus was stained by DAPI (blue). Scale bar = 50 μm. The bar graph shows area fractions of α-SMA (%) in the sham and UUO kidneys from ten randomly chosen, non-overlapping fields at a magnification of 400× ( n = 10 per group). ( B ) The sections were stained with Picro Sirius Red. Scale bar = 100 μm. The bar chart shows Picro Sirius Red (+) areas (%) in the sham and UUO kidneys from ten randomly chosen, non-overlapping fields at a magnification of 200× ( n = 10 per group). ( C ) Representative Western blot analysis of α-SMA and type I collagen expression in the kidneys from sham- and UUO-operated mice treated with Veh or HKL. The bar graph shows the densitometric quantification presented as the relative ratio of each protein to GAPDH. The relative ratio measured in the kidneys from sham-operated mice treated with Veh is arbitrarily presented as 1. Data are expressed as mean ± SD. *** p < 0.001 versus Veh or HKL; # p < 0.05 and ### p < 0.001 versus UUO treatment with Veh. Sham, sham-operated mice; UUO, unilateral ureteral obstruction; Veh, vehicle; HKL, honokiol; α-SMA, α-smooth muscle actin; DAPI, 4’,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The tissue samples were incubated with either a rat anti-mouse F4/80 (14-4801-82; dilution 1:200; eBioscience, San Diego, CA, USA) or a mouse anti-α-smooth muscle actin (α-SMA; dilution 1:200; BD Biosciences, Allschwil, Switzerland).

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Sirtuin 3 Activation by Honokiol Decreases Unilateral Ureteral Obstruction-Induced Renal Inflammation and Fibrosis via Regulation of Mitochondrial Dynamics and the Renal NF-κB-TGF-β1/Smad Signaling Pathway

doi: 10.3390/ijms21020402

Figure Lengend Snippet: Effect of honokiol on transforming growth factor-β1 (TGF-β1)-induced fibroblast activation and extracellular matrix protein expression by regulation of the TGF-β1/Smad signaling pathway in NRK-49F cells ( A ) Representative Western blot for α-SMA, fibronectin, and type I collagen from NRK-49F cells treated with vehicle (Veh) or TGF-β1 (2 ng/mL), with or without honokiol (HKL) at indicated doses (0.1, 1, and 10 μM). Treatment with TGF-β1 (2 ng/mL) for 24 h increased the expression of fibrotic markers. The expression of α-smooth muscle actin (α-SMA), fibronectin, and type I collagen decreased after HKL treatment in a dose-dependent manner. The bar graph shows the densitometric quantification presented as the relative ratio of each protein to GAPDH. Data are presented as mean ± SD. ( B ) Representative Western blot for phospho-Smad2 and phospho-Smad3 expression from NRK-49F cells treated with vehicle (Veh) or TGF-β1 (2 ng/mL), with or without honokiol (HKL) at indicated doses (0.1, 1, and 10 μM). TGF-β1 (2 ng/mL) treatment for 30 min increased the expression of phospho-Smad2 and phospho-Smad3. Measurement of phospho-Smad2 and phospho-Smad3 protein expression of densitometry is presented as the relative ratio to Smad2/3 protein expression. The bar graph shows the densitometric quantification presented as the relative ratio of each protein to Smad2/3. Data are represented as mean ± SD. * p < 0.05 and *** p < 0.001 versus Veh or HKL; # p < 0.05, ## p < 0.01 and ### p < 0.001 versus TGF-β1 treatment. Veh, vehicle; HKL, honokiol; TGF-β1, transforming growth factor-β1; α-SMA, α-smooth muscle actin.

Article Snippet: The tissue samples were incubated with either a rat anti-mouse F4/80 (14-4801-82; dilution 1:200; eBioscience, San Diego, CA, USA) or a mouse anti-α-smooth muscle actin (α-SMA; dilution 1:200; BD Biosciences, Allschwil, Switzerland).

Techniques: Activation Assay, Expressing, Western Blot