Journal: Frontiers in Endocrinology

Article Title: Chaihu-Wendan Decoction alleviates obesity through PTEN-mediated uncoupling of metabolic signaling and macrophage activation

doi: 10.3389/fendo.2026.1779657

Figure Lengend Snippet: Effect of CHWD on protein expression of STAT3, BCL2, HIF1A, AKT, and mTOR in omental tissues of different groups of mice. Representative Western blots (A) and densitometric analysis of (B) STAT3, (C) HIF1A, (D) AKT, (E) mTOR, and (F) BCL2. Data are expressed as mean ± SD (n = 3). Statistical differences are based on P < 0.05 (*) and P < 0.01 (**) vs. Control group and based on P < 0.05 (#) and P < 0.01 (##) vs. Model group.

Article Snippet: Antibodies and Kits: Primary antibodies against STAT3 (AF6294, 1:1000), HIF1A (AF1009, 1:1000), mTOR (AF6308, 1:1000), p-mTOR (AF3308, 1:1000), BCL2 (AF6139, 1:1000), p-IRS1 (AF3272, 1:1000), p-PI3K (AF3242, 1:1000), and PTEN (AF5447, 1:1000) were purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Western Blot, Control

Journal: Scientific Reports

Article Title: Peritoneal MSCs-derived exosomes suppress CCL24 synthesis through miR-320d delivery contributing to the improvement of peritoneal dialysis-associated fibrosis

doi: 10.1038/s41598-026-42489-w

Figure Lengend Snippet: The effects of HpMSCs-Exo and miR-320d on CCL24 expression in macrophages. ( A ) The miRNA level in THP1-derived macrophages treated with or without HpMSCs-derived exosomes (HpMSCs-Exo) was measured with qRT-PCR assay. ( B ) After incubating with HpMSCs-Exo with or without FITC-labeled miR-320d, FITC in macrophages was observed under a fluorescence microscope. Scale bars: 20 μm. C-D. THP1-derived macrophages were treated with or without HpMSCs-Exo together with miRNA inhibitor NC (inhibitor NC) or miR-320d inhibitor (miR-320d in). ( C ) qRT-PCR assay was conducted to detect the level of miR-320d in macrophages. ( D ) The protein levels of p-STAT3, STAT3, and CCL24 in macrophages with different treatments were measured with western blotting (quantified in the right). Values are the mean ± SD; Student’s t-test (Panel: A) or One-way ANOVA with Tukey’s post hoc test (Panel: C-D); * p < 0.05, ** p < 0.01, ns p > 0.05. Mφ: THP1-derived macrophages; HpMSCs: human peritoneal mesenchymal stem cells.

Article Snippet: Immunoprecipitation reactions were performed by using antibodies against STAT3 (CST, 12640), and rabbit IgG (CST, 2729).

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Labeling, Fluorescence, Microscopy, Western Blot

Journal: Scientific Reports

Article Title: Peritoneal MSCs-derived exosomes suppress CCL24 synthesis through miR-320d delivery contributing to the improvement of peritoneal dialysis-associated fibrosis

doi: 10.1038/s41598-026-42489-w

Figure Lengend Snippet: The effects of KLF7 on HpMSCs-Exo or miR-320d-mediated STAT3/CCL24 pathway inhibition in macrophages. ( A ). After transfection with luciferase reporter plasmids containing wild or mutated KLF7 mRNA 3’ UTR sequence (displayed in the left), which binds to miR-320d in TargetScan, THP1-derived macrophages were treated with miRNA mimics NC (mimics NC) or miR-320d mimics (miR-320d mi). Then the luciferase activity in macrophages was measured. ( B ). Western blotting was conducted to measure the protein expression of KLF7 in macrophages treated with or without HpMSCs-derived exosomes (HpMSCs-Exo) in the presence of miR-320d mimics or miR-320d inhibitor (quantified in the right). ( C - D ). THP1-derived macrophages were transfected with miRNA mimics NC or miR-320d mimics together with vector or KLF7 overexpression plasmids. Western blotting was adopted to detect the protein expression of KLF7 and the phosphorylation level of STAT3 ( C ) and the protein level of CCL24 ( D ) in macrophages. E-F. After being transfected with vectors or KLF7 overexpression plasmids, THP1-derived macrophages were incubated with or without HpMSCs-Exo. Western blotting was used to measure the protein expression of KLF7 and the phosphorylation level of STAT3 ( E ) and the protein level of CCL24 ( F ) in macrophages. Values are the mean ± SD; Student’s t-test (Panel: A) or One-way ANOVA with Tukey’s post hoc test (Panel: B-F); * p < 0.05, ** p < 0.01, ns p > 0.05. Mφ: THP1-derived macrophages; HpMSCs: human peritoneal mesenchymal stem cells.

Article Snippet: Immunoprecipitation reactions were performed by using antibodies against STAT3 (CST, 12640), and rabbit IgG (CST, 2729).

Techniques: Inhibition, Transfection, Luciferase, Sequencing, Derivative Assay, Activity Assay, Western Blot, Expressing, Plasmid Preparation, Over Expression, Phospho-proteomics, Incubation

Journal: Scientific Reports

Article Title: Peritoneal MSCs-derived exosomes suppress CCL24 synthesis through miR-320d delivery contributing to the improvement of peritoneal dialysis-associated fibrosis

doi: 10.1038/s41598-026-42489-w

Figure Lengend Snippet: The effects of modified HpMSCs-Exo loaded with miR-320d on KLF7/STAT3/CCL24 pathway, MMT, and PD-related fibrosis. ( A ). Exosomes were separated from the supernatant of HpMSCs transfected with miRNA mimics NC or miR-320d mimics, and then the abundance of miR-320d in native exosomes (Exo-C) and modified exosomes (Exo-M) was measured with qRT-PCR assay. B-C. THP1-derived macrophages were incubated with Exo-C or Exo-M. And then qRT-PCR ( B ) and western blotting ( C ) were adopted to measure the level of miR-320d and the protein expression of KLF7 and CCL24, and the phosphorylation level of STAT3 (quantified in the right). ( D ). Western blotting was used to measure the phosphorylation level of p38 and the protein content of E-cadherin, α-SMA, FN, and collagen I in HPMCs treated with the supernatant of THP1-derived macrophages stimulated with medium (Mφ-S) or Exo-C (Mφ Exo−C -S), or Exo-M (Mφ Exo−M -S) (quantified in the right). ( E - F ). Rat primary peritoneal macrophages were incubated with Exo-C or Exo-M. And then qRT-PCR ( E ) and western blotting ( F ) were adopted to measure the level of miR-320d and the protein expression of KLF7 and CCL24, and the phosphorylation level of STAT3 (quantified in the right). ( G ). Western blotting was used to measure the phosphorylation level of p38 and the protein content of E-cadherin, α-SMA, FN, and collagen I in RPMCs treated with the supernatant of rat primary peritoneal macrophages stimulated with medium (PMφ-S) or Exo-C (PMφ Exo−C -S), or Exo-M (PMφ Exo−M -S) (quantified in the right). ( H - L ). A PD-associated peritoneal injury rat model was established with intraperitoneal injection of 4.25% glucose PD solution and LPS. And then PD rats were divided into three groups and subjected to different treatments, including tail vein injection of PBS or Exo-C or Exo-M ( n = 6/groups). Meanwhile, 6 control rats were only exposed to intraperitoneal and tail vein injection of PBS. Then the peritoneal tissues were collected. ( H ). HE staining was conducted to measure the thickness of the peritoneal membrane in control rats and PD rats with different treatments (quantified in the right). Scale bars: 100 μm. ( I ). Masson staining of peritoneal tissues from rats of different groups. Scale bars: 100 μm. ( J ). The level of miR-320d in the peritoneal tissues from different rats was detected by qRT-PCR. ( K ). ELISA kit was used to measure the content of CCL24 in peritoneal tissues. ( L ). Western blotting was conducted to detect the phosphorylation level of p38 and the protein content of E-cadherin, α-SMA, FN, and collagen I in the peritoneal tissues (quantified in the right). Values are the mean ± SD; Student’s t-test (Panel: A) or One-way ANOVA with Tukey’s post hoc test (Panel: B-L); * p < 0.05, ** p < 0.01, ns p > 0.05. Mφ: THP1-derived macrophages; PMφ: Rat primary peritoneal macrophages; MMT: Mesothelial-to-mesenchymal transition; HpMSCs: human peritoneal mesenchymal stem cells; HPMC: human peritoneal mesothelial cells; FN: fibronectin; PD: peritoneal dialysis.

Article Snippet: Immunoprecipitation reactions were performed by using antibodies against STAT3 (CST, 12640), and rabbit IgG (CST, 2729).

Techniques: Modification, Transfection, Quantitative RT-PCR, Derivative Assay, Incubation, Western Blot, Expressing, Phospho-proteomics, Injection, Control, Staining, Membrane, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Peritoneal MSCs-derived exosomes suppress CCL24 synthesis through miR-320d delivery contributing to the improvement of peritoneal dialysis-associated fibrosis

doi: 10.1038/s41598-026-42489-w

Figure Lengend Snippet: The mechanism diagram by which native or modified HpMSCs-Exo alleviate PD-related fibrosis progression. HpMSCs-Exo, especially modified HpMSCs-Exo loaded with miR-320d, can deliver miR-320d into macrophages, resulting in the upregulation of intracellular miR-320d levels. The enriched miR-320d subsequently binds to KLF7 mRNA and suppresses its protein expression, thereby inhibiting KLF7-dependent STAT3 activation and downstream CCL24 synthesis in macrophages. Reduced secretion of CCL24 attenuates the CCR3/p38 MAPK pathway activation and ameliorates MMT, ultimately alleviating PD-associated fibrosis.

Article Snippet: Immunoprecipitation reactions were performed by using antibodies against STAT3 (CST, 12640), and rabbit IgG (CST, 2729).

Techniques: Modification, Expressing, Activation Assay

Journal: Biomedical Reports

Article Title: Ethyl acetate fraction of Curcuma longa leaves suppresses IL-6-induced STAT3 activation via ERK signaling in Hep3B cells

doi: 10.3892/br.2026.2108

Figure Lengend Snippet: Ethyl acetate fraction of CL-E inhibits IL-6-induced STAT3 activation in Hep3B cells. (A) Hep3B cells stably expressing STAT3-responsive luciferase reporter (pSTAT3-Luc) were treated with IL-6 (10 ng/ml) for 12 h in the presence or absence of CL-E or an aqueous fraction of C. longa L. leaves extract at concentrations of 10, 30, and 60 µg/ml. Luciferase activity was measured according to the manufacturer's instructions. Data are presented as relative luciferase activity normalized to the untreated control. (B) Hep3B cells were seeded in 96-well plates and treated with each fraction for 24 h at the indicated concentrations. Cell viability was assessed using the MTT assay. Results are expressed as the percentage of viable cells relative to the untreated control. Data represent the mean ± SE (n≥3). * P<0.05 and ** P<0.01 vs. the IL-6 alone group; ## P<0.01 indicates a significant difference between the 10 and 60 µg/ml CL-E doses. CL-E, Curcuma longa L. extract; IL-6, interleukin 6; STAT3, signal transducer and activator of transcription 3.

Article Snippet: Membranes were blocked with 5% (w/v) non-fat dry milk in TBS-T (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4 ̊C with primary antibodies against p-STAT3 (Tyr705; 1:1,000; CST #9145), p-STAT3 (Ser727; 1:1,000; CST #9134), t-STAT3 (1:1,000; CST #4904), p-ERK (1:1,000; CST #4370), t-ERK (1:1,000; CST #4695) and β-actin (1:2,000; CST #4967).

Techniques: Activation Assay, Stable Transfection, Expressing, Luciferase, Activity Assay, Control, MTT Assay

Journal: Biomedical Reports

Article Title: Ethyl acetate fraction of Curcuma longa leaves suppresses IL-6-induced STAT3 activation via ERK signaling in Hep3B cells

doi: 10.3892/br.2026.2108

Figure Lengend Snippet: CL-E inhibits IL-6-induced STAT3 phosphorylation and nuclear translocation in Hep3B cells. (A) Hep3B cells were treated with IL-6 (10 ng/ml) for the indicated time periods (0-60 min). Total cell lysates were analyzed by western blotting using antibodies against P-STAT3 (Tyr705), P-STAT3 (Ser727) and total STAT3. (B) Cells were pretreated with the CL-E at 10, 30 and 60 µg/ml for 1 h and then stimulated with IL-6 (10 ng/ml) for 30 min. Proteins were extracted and analyzed by western blotting as described above. (C) STAT3 nuclear translocation was assessed by immunofluorescence microscopy. Hep3B cells were cultured on Nunc Lab-Tek II 8-well chamber slides and treated with IL-6 (10 ng/ml) for 4 h in the absence or presence of CL-E (60 µg/ml) or genistein (60 µM, positive control). Cells were fixed, permeabilized and stained with anti-STAT3 antibody followed by FITC-conjugated secondary antibody. Subcellular localization of STAT3 was visualized using fluorescence microscopy (Scale bar: 20 µm). CL-E, Curcuma longa L. extract; IL-6, interleukin 6; STAT3, signal transducer and activator of transcription 3; P-, phosphorylated.

Article Snippet: Membranes were blocked with 5% (w/v) non-fat dry milk in TBS-T (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4 ̊C with primary antibodies against p-STAT3 (Tyr705; 1:1,000; CST #9145), p-STAT3 (Ser727; 1:1,000; CST #9134), t-STAT3 (1:1,000; CST #4904), p-ERK (1:1,000; CST #4370), t-ERK (1:1,000; CST #4695) and β-actin (1:2,000; CST #4967).

Techniques: Phospho-proteomics, Translocation Assay, Western Blot, Immunofluorescence, Microscopy, Cell Culture, Positive Control, Staining, Fluorescence

Journal: Biomedical Reports

Article Title: Ethyl acetate fraction of Curcuma longa leaves suppresses IL-6-induced STAT3 activation via ERK signaling in Hep3B cells

doi: 10.3892/br.2026.2108

Figure Lengend Snippet: ERK signaling pathway contributes to CL-E-mediated regulation of IL-6-induced STAT3 activation. (A) Hep3B cells were stimulated with IL-6 (10 ng/ml) for the indicated times (0, 5, 10, 30 and 60 min). Protein lysates underwent western blotting analysis using antibodies against P-ERK1/2 and total ERK1/2 to evaluate time-dependent ERK1/2 activation. (B) Cells were pretreated with CL-E at 10, 30 and 60 µg/ml for 1 h, followed by IL-6 stimulation (10 ng/ml) for 30 min. ERK1/2 phosphorylation was analyzed by western blotting analysis. (C) To investigate the role of ERK in STAT3 Ser727 phosphorylation, cells were treated with CL-E (60 µg/ml), PKC inhibitor bisindolylmaleimide II (20 µM), and/or the MEK1/2 inhibitor U0126 (20 µM) for 1 h in the absence of IL-6. Phosphorylation of STAT3 (Ser727) and total STAT3 were examined by western blotting analysis. (D) To determine whether ERK activation contributes to regulation of STAT3 Tyr705 phosphorylation, cells were pretreated with CL-E, U0126 and/or bisindolylmaleimide II (20 µM) for 1 h before IL-6 stimulation (10 ng/ml, 30 min). STAT3 Tyr705 phosphorylation was analyzed by western blotting. CL-E, Curcuma longa L. extract; IL-6, interleukin 6; STAT3, signal transducer and activator of transcription 3; P-, phosphorylated.

Article Snippet: Membranes were blocked with 5% (w/v) non-fat dry milk in TBS-T (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4 ̊C with primary antibodies against p-STAT3 (Tyr705; 1:1,000; CST #9145), p-STAT3 (Ser727; 1:1,000; CST #9134), t-STAT3 (1:1,000; CST #4904), p-ERK (1:1,000; CST #4370), t-ERK (1:1,000; CST #4695) and β-actin (1:2,000; CST #4967).

Techniques: Activation Assay, Western Blot, Phospho-proteomics