polydatin  (Macklin Inc)

Journal: Animal Models and Experimental Medicine

Article Title: Polydatin alleviates mitochondrial damage and apoptosis of lung epithelial cells by inhibiting toll‐like receptor 4‐dependent macrophage activation in asthma

doi: 10.1002/ame2.70100

Figure Lengend Snippet: Polydatin relieves ROS (reactive oxygen species) and inflammation in asthmatic mice. To investigate asthma pathology, we developed an asthma mouse model via sensitization of BALB/c mice with ovalbumin (OVA). After initial OVA exposure, animals received intraperitoneal injections of polydatin prior to subsequent aerosolized OVA challenges. (A) Structural representation of polydatin. (B) Experimental protocol diagram illustrating timeline and treatment schedules. (C) Lung resistance alterations in response to methacholine stimulation ( n = 8 per group). (D) Quantification of eosinophils (identified as CD45.2 + Siglec‐F + cells) within bronchoalveolar lavage fluid (BALF) using flow cytometry. (E) Representative histological lung sections stained with hematoxylin–eosin (HE) and periodic acid–Schiff (PAS). (F) Measurement of cytokine concentrations in BALF using ELISA (enzyme‐linked immunosorbent assay) assays ( n = 8 per group). (G) Fluorescent microscopic images demonstrating ROS (reactive oxygen species) production via dihydroethidium (DHE) staining in lung tissues. (H) WB (Western blotting) quantification of NOX2 (NADPH oxidase 2) and NOX4 (NADPH oxidase 4) protein expression in pulmonary tissues. (I) Identification of apoptotic cells using TUNEL (terminal deoxynucleotidyl transferase dUTP nick‐end labeling) staining in lung sections with corresponding quantitative analysis of positive cells ( n = 3). (J) WB detection of apoptosis‐related proteins within lung homogenates. (K) Immunohistochemical staining depicting the expression and localization of cleaved caspase‐3 and cleaved caspase‐9 proteins ( n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Prior to LPS and ATP stimulation, cells were pretreated for 1 h with polydatin (50 or 100 μmol/L), CLI‐095 (a TLR4‐specific inhibitor, CAS: 243984‐11‐4, 1 μg/mL, InvivoGen), or A438079 (a P2X7R‐specific antagonist, CAS: 899507‐36‐9, 10 μmol/L, Abcam).

Techniques: Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, TUNEL Assay, Immunohistochemical staining

Journal: Animal Models and Experimental Medicine

Article Title: Polydatin alleviates mitochondrial damage and apoptosis of lung epithelial cells by inhibiting toll‐like receptor 4‐dependent macrophage activation in asthma

doi: 10.1002/ame2.70100

Figure Lengend Snippet: Polydatin suppresses TLR4 (toll‐like receptor 4)/P2X7R/NLRP3 (NOD‐like receptor protein) activation in OVA (ovalbumin)–induced asthma. WB (Western blotting) measured (A) TLR4, IκBα, NF‐κB (nuclear factor kappa B) P65, and phosphorylation of IκBα and NF‐κB P65, and (B) P2X7R, NLRP3, ASC, and caspase‐1 in lung tissues. (C) Co‐expression of F4/80 (green) and (D) P2X7R (green) with TLR4 (red) in mouse lung tissues was assessed via immunofluorescence staining combined ( n = 3). (E) ELISA (enzyme‐linked immunosorbent assay) and real‐time PCR (polymerase chain reaction) were used to measure IL‐1β (interleukin‐1β) and IL‐18 protein levels in BALF (bronchoalveolar lavage fluid) and mRNA (messenger RNA) expression ( n = 8), and Dunnett's post hoc test (A–D) or Wilcoxon rank‐sum test (E) was performed subsequently. * p < 0.05 and ** p < 0.01.

Article Snippet: Prior to LPS and ATP stimulation, cells were pretreated for 1 h with polydatin (50 or 100 μmol/L), CLI‐095 (a TLR4‐specific inhibitor, CAS: 243984‐11‐4, 1 μg/mL, InvivoGen), or A438079 (a P2X7R‐specific antagonist, CAS: 899507‐36‐9, 10 μmol/L, Abcam).

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Expressing, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, RNA Expression

Journal: Animal Models and Experimental Medicine

Article Title: Polydatin alleviates mitochondrial damage and apoptosis of lung epithelial cells by inhibiting toll‐like receptor 4‐dependent macrophage activation in asthma

doi: 10.1002/ame2.70100

Figure Lengend Snippet: Polydatin inhibits IL‐1β (interleukin‐1β) and IL‐18 by blocking the TLR4 (toll‐like receptor 4)/P2X7R pathway in macrophages. (A) Intracellular calcium concentrations were analyzed utilizing Fluo 3‐AM fluorescent staining ( n = 3). (B) WB (Western blotting) was performed to quantify the levels of TLR4, IκBα, NF‐κB (nuclear factor kappa B) p65, phosphorylated forms of IκBα and NF‐κB p65, P2X7R, NLRP3 (NOD‐like receptor protein) inflammasome components, IL‐1β, and IL‐18 ( n = 3). (C) Macrophage‐derived protein secretion levels of IL‐1β and IL‐18 were evaluated using ELISA (enzyme‐linked immunosorbent assay). (A and B) or Wilcoxon rank‐sum testing (C). * p < 0.05 and ** p < 0.01.

Article Snippet: Prior to LPS and ATP stimulation, cells were pretreated for 1 h with polydatin (50 or 100 μmol/L), CLI‐095 (a TLR4‐specific inhibitor, CAS: 243984‐11‐4, 1 μg/mL, InvivoGen), or A438079 (a P2X7R‐specific antagonist, CAS: 899507‐36‐9, 10 μmol/L, Abcam).

Techniques: Blocking Assay, Staining, Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: Animal Models and Experimental Medicine

Article Title: Polydatin alleviates mitochondrial damage and apoptosis of lung epithelial cells by inhibiting toll‐like receptor 4‐dependent macrophage activation in asthma

doi: 10.1002/ame2.70100

Figure Lengend Snippet: Polydatin suppresses ROS (reactive oxygen species) production and apoptosis of ECs (epithelial cells) by blockage of the TLR4 (toll‐like receptor 4)/P2X7R pathway in macrophages. (A) Intracellular ROS levels in BEAS‐2B cells were determined through DCFH‐DA fluorescence staining (10 μmol/L). (B) Alterations in MMP (mitochondrial membrane potential) were assessed using JC‐1 dye staining. (C) WB (Western blotting) analysis quantified protein expression of NOX2 (NADPH oxidase 2) and NOX4 (NADPH oxidase 4), and proteins related to apoptosis in BEAS‐2B ECs (epithelial cells). (D) Flow cytometry quantified apoptosis based on the proportion of annexin‐V + /7‐AAD + positive cells ( n = 3). (E) Additionally, WB was used to determine apoptosis‐related protein levels in BEAS‐2B cells. Histogram results represent the mean ± SEM (standard error of the mean) from three independent replicates. * p < 0.05 and ** p < 0.01.

Article Snippet: Prior to LPS and ATP stimulation, cells were pretreated for 1 h with polydatin (50 or 100 μmol/L), CLI‐095 (a TLR4‐specific inhibitor, CAS: 243984‐11‐4, 1 μg/mL, InvivoGen), or A438079 (a P2X7R‐specific antagonist, CAS: 899507‐36‐9, 10 μmol/L, Abcam).

Techniques: Fluorescence, Staining, Membrane, Western Blot, Expressing, Flow Cytometry

Journal: Animal Models and Experimental Medicine

Article Title: Polydatin alleviates mitochondrial damage and apoptosis of lung epithelial cells by inhibiting toll‐like receptor 4‐dependent macrophage activation in asthma

doi: 10.1002/ame2.70100

Figure Lengend Snippet: Schematic diagram of the mechanisms underlying the alleviation of OVA (ovalbumin)–induced asthma by polydatin. The alleviation of asthma by polydatin is dependent on the blockage of the TLR4 (toll‐like receptor 4)/P2X7R synergy in macrophages. The blockage of the TLR4/P2X7R synergy results in decreased release and secretion of IL‐1β (interleukin‐1β) and IL‐18In ECs, low IL‐1β and IL‐18 levels inhibit mitochondrial damage and apoptosis.

Article Snippet: Prior to LPS and ATP stimulation, cells were pretreated for 1 h with polydatin (50 or 100 μmol/L), CLI‐095 (a TLR4‐specific inhibitor, CAS: 243984‐11‐4, 1 μg/mL, InvivoGen), or A438079 (a P2X7R‐specific antagonist, CAS: 899507‐36‐9, 10 μmol/L, Abcam).

Techniques:

Journal: MethodsX

Article Title: An accessible HPLC-DAD method for the direct detection of acrolein-trapping compounds in complex plant matrices ☆

doi: 10.1016/j.mex.2025.103747

Figure Lengend Snippet: Chromatographic profile of the ten-component mixture, with detection at a wavelength of 280 nm. The upper chromatogram corresponds to the negative control (0 mM acrolein), while the lower chromatogram represents the mixture following pretreatment with 5 mM acrolein. 1: protocatechuic acid, 2: esculin, 3: catechin, 4: epicatechin, 5: coumaric acid, 6: piceid, 7: rutin, 8: phloridzin, 9: resveratrol, 10: phloretin. Acrolein-trapping compounds are highlighted in red.

Article Snippet: 2,4-Dinitrophenylhydrazine hydrochloride (DNPH), aminoguanidine hydrochloride, esculin, piceid and phloretin were obtained from TCI Chemicals.

Techniques: Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Lactate-driven lactylation of HNRNPA1 orchestrates PKM2 splicing and glycolytic reprogramming in bladder cancer

doi: 10.1186/s13046-025-03591-5

Figure Lengend Snippet: Polydatin inhibits HNRNPA1 lactylation and its biological activity in bladder cancer. A Binding free energy of ten candidate compounds docked to the HNRNPA1 active pocket. B Chemical structure of polydatin. C Docking visualization of polydatin within the active pocket of HNRNPA1. D IC 50 values of polydatin in the EJ-1 cell line, as determined by CCK8 assay. E EJ-1 cells were lysed and immunoprecipitated using an anti-HNRNPA1 antibody, followed by detection of HNRNPA1-K350 lactyl lysine. F Colony formation assays were performed to assess the proliferative capacity of EJ-1 cells following 24-h treatment with 30 μM polydatin or 20 mM NaLa. G-H Tumor gross images and tumor weights were assessed in nude mice after subcutaneous injection of EJ-1 cells. Tumors were harvested on Day 19 after subcutaneous implantation. Statistical analyses were performed with unpaired two-tailed Student’s t-test ( n = 5). I Tumor volume measurements throughout the experiment. J Immunohistochemical staining visualized PKM2 levels in xenograft tumors. Scale bars: 100 μm (left panel), 50 μm (right panel). K Schematic representation of the proposed mechanism of HNRNPA1 lactylation. * p < 0.05, ** p < 0.01

Article Snippet: Cells were treated with polydatin (27208–80-6, TargetMol, USA) for 24 h to examine cell proliferation.

Techniques: Activity Assay, Binding Assay, CCK-8 Assay, Immunoprecipitation, Injection, Two Tailed Test, Immunohistochemical staining, Staining