Journal: Drug Design, Development and Therapy

Article Title: Treatment with a PPAR-γ Agonist Protects Against Hyperuricemic Nephropathy in a Rat Model

doi: 10.2147/DDDT.S247091

Figure Lengend Snippet: RGTZ abrogates TGF-β-Smad3 signaling in the kidneys of HN rats. ( A ) Kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against TGF-β1 or glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( B ) Expression levels of TGF-β1 were quantified by densitometry and normalized to GAPDH. ( C ) The kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against p-Smad3, Smad3, or GAPDH. ( D ) Expression levels of p- Smad3 were quantified by densitometry and normalized to Smad3. RGTZ, rosiglitazone. Data are represented as the mean ± SEM. *p < 0.05 vs sham group; #p< 0.05 vs sham + RGTZ group. **p < 0.05 vs HN group.

Article Snippet: Antibodies to Smad3 and p-Smad3 were purchased from Hangzhou HuaAn Biotechnology Co., Ltd. (Hangzhou, China).

Techniques: Western Blot, Expressing

Journal: The FASEB Journal

Article Title: Mitigation of Ferroptosis in Diabetic Kidney Disease Through Mesenchymal Stem Cell Intervention via the Smad2/3/METTL3/S1PR1 Axis

doi: 10.1096/fj.202403207r

Figure Lengend Snippet: FIGURE 1 | Effects of MSC intervention on renal pathology, inflammation, Smad2/3 activation, and ferroptosis in DKD Mice: (A, B) Schematic representation of the modeling and intervention process in C57 and db/db mice. (C) Blood glucose levels monitored across the experimental groups. (D) Comparison of Kidney weight/Body weight ratios between groups. (E) H&E staining to assess renal pathological changes, and Masson staining to quantify renal fibrosis. Representative histopathological images are provided, with Masson staining quantification depicted on the right. (F) ELISA detection of IL-6 and TNF-α levels as markers of inflammation. (G) Western blot analysis of Smad2, Smad3, and their phosphorylated forms (p- Smad2, p-Smad3), with β-Actin as the loading control. (H) Immunohistochemistry to visualize p-Smad2/3 nuclear translocation, with magnification at 200× and nuclear staining shown in blue. (I) Western blot analysis of ferroptosis-related proteins (ACSL4, GPX4, and SLC7A11), using β-Actin as the loading control. Control Group: C57 Mice on a standard diet, administered an equal volume of physiological saline via tail vein injection. DKD Group: db/db mice with an established T2DM model, receiving physiological saline via tail vein injection. DKD + MSCs Group: db/db mice with an established T2DM model, treated with MSCs (1 × 106 cells/0.2 mL per mouse) via weekly tail vein infusion. Interventions were conducted over an 8- week period. Each group consisted of 6 mice, and all mice participated in and underwent all experimental assessments. All in vitro experiments were performed with at least three independent replicates (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).

Article Snippet: Primary antibodies targeting Smad3 (Affinity, 1:100) and Smad2 (Huabio, 1:100) were applied and incubated overnight at 4°C.

Techniques: Activation Assay, Comparison, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Immunohistochemistry, Translocation Assay, Saline, Injection, In Vitro

Journal: The FASEB Journal

Article Title: Mitigation of Ferroptosis in Diabetic Kidney Disease Through Mesenchymal Stem Cell Intervention via the Smad2/3/METTL3/S1PR1 Axis

doi: 10.1096/fj.202403207r

Figure Lengend Snippet: FIGURE 3 | MSCs inhibit the activation of the Smad2/3 pathway and ferroptosis. (A, B) Western blot analysis of Smad2, Smad3, and their phos- phorylated forms (p-Smad2, p-Smad3), with β-Actin as the loading control. (C) Immunofluorescence analysis of Smad2/3 nuclear localization. Nuclei stained white, while Smad2/3 are labeled green using goat anti-rabbit IgG H&L (Alexa Fluor 488) secondary antibody. (D) Western blot analysis of ferroptosis-related proteins (ACSL4, GPX4, and SLC7A11), with β-Actin as the loading control. Grouping: Control Group: MPC5 Cells cultured in standard medium. HG Group: MPC5 Cells treated with 25 μmol/L glucose in standard medium for 72 h. HG + MSCs Group: MPC5 Cells co-cultured with MSCs in medium containing 25 μmol/L glucose for 72 h. All in vitro experiments were performed with at least three independent replicates (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).

Article Snippet: Primary antibodies targeting Smad3 (Affinity, 1:100) and Smad2 (Huabio, 1:100) were applied and incubated overnight at 4°C.

Techniques: Activation Assay, Western Blot, Control, Immunofluorescence, Staining, Labeling, Cell Culture, In Vitro