Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Olaparib Blocks Oxidative Stress‐Dependent NLRP3 Inflammasome in Pediatric Acute Lung Injury by Metabolic Reprogramming via AMPK ‐Driven Autophagic Flux

doi: 10.1002/kjm2.70275

Figure Lengend Snippet: Olaparib activates AMPK‐dependent metabolic and autophagic responses in macrophages. (A) Cell viability of PMA‐differentiated THP‐1 macrophages treated with Olaparib (0–50 μM) was assessed using the CCK‐8 assay. * p < 0.05 versus control group. (B) NAD + /NADH ratio was measured in PMA‐differentiated THP‐1 macrophages under control, LPS/ATP, LPS/ATP + Olaparib (5 μM), LPS/ATP + Olaparib (10 μM), LPS/ATP + Olaparib (20 μM), and LPS/ATP + Olaparib (20 μM) + compound C (10 μM) conditions. (C) AMP/ATP ratio in THP‐1 macrophages across the indicated experimental groups. (D–E) Protein expression levels of p‐AMPK, AMPK, p‐ULK1, and ULK1 in THP‐1 macrophages assessed by Western blot. (F–G) LC3‐II/LC3‐I ratio and p62 protein levels determined by Western blot analysis. (H–I) Autophagic flux in control, LPS/ATP, LPS/ATP + Olaparib (20 μM), and LPS/ATP + Olaparib (20 μM) + compound C (10 μM) groups evaluated using the mRFP–GFP–LC3 tandem fluorescent reporter system. Data are presented as mean ± standard deviation from three independent experiments. &&& p < 0.001 versus control group; * p < 0.05, ** p < 0.01, *** p < 0.001 versus LPS/ATP group; ### p < 0.001 versus LPS/ATP + Olaparib (20 μM) group.

Article Snippet: In the combination group, compound C (20 mg/kg; 99.10% purity; HY‐13418A, MedChemExpress) was co‐administered with Olaparib (100 mg/kg) 1 h after LPS injection.

Techniques: CCK-8 Assay, Control, Expressing, Western Blot, Analysis, Standard Deviation

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Olaparib Blocks Oxidative Stress‐Dependent NLRP3 Inflammasome in Pediatric Acute Lung Injury by Metabolic Reprogramming via AMPK ‐Driven Autophagic Flux

doi: 10.1002/kjm2.70275

Figure Lengend Snippet: Olaparib suppresses oxidative stress and NLRP3 activation via AMPK‐dependent autophagy in ALI mice. (A) NAD + /NADH ratio in lung tissues from control, LPS, LPS + Olaparib (50 mg/kg), LPS + Olaparib (100 mg/kg), and LPS + Olaparib (100 mg/kg) + compound C (20 mg/kg) groups. (B–C) Western blot analysis of p‐AMPK/AMPK and LC3‐II/LC3‐I ratios, along with p62 protein levels in lung tissues. (D–E) Immunohistochemical staining and quantification of 4‐HNE expression. (F) Measurement of MDA levels in lung tissues. (G) Levels of IL‐1β and IL‐18 in BALF determined by ELISA. (H–I) Western blot analysis of NLRP3, ASC, and cleaved caspase‐1 protein expression. Data are presented as mean ± standard deviation ( n = 6). &&& p < 0.001 versus sham group; * p < 0.05, ** p < 0.01, *** p < 0.001 versus LPS group; ## p < 0.01, ### p < 0.001 versus LPS + Olaparib (100 mg/kg) group.

Article Snippet: In the combination group, compound C (20 mg/kg; 99.10% purity; HY‐13418A, MedChemExpress) was co‐administered with Olaparib (100 mg/kg) 1 h after LPS injection.

Techniques: Activation Assay, Control, Western Blot, Analysis, Immunohistochemical staining, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Olaparib Blocks Oxidative Stress‐Dependent NLRP3 Inflammasome in Pediatric Acute Lung Injury by Metabolic Reprogramming via AMPK ‐Driven Autophagic Flux

doi: 10.1002/kjm2.70275

Figure Lengend Snippet: Olaparib alleviates pulmonary injury and inflammatory cell infiltration in ALI mice. (A) Representative H&E‐stained lung sections from control, LPS, LPS + Olaparib (50 mg/kg), LPS + Olaparib (100 mg/kg), and LPS + Olaparib (100 mg/kg) + compound C (20 mg/kg) groups. (B) Quantification of lung injury score. (C) Lung wet/dry (W/D) ratio assessment. (D–E) Immunohistochemical staining and quantification of F4/80‐positive areas in lung tissues. (F) MPO activity in BALF. (G) MPO activity in lung tissue homogenates. Data are presented as mean ± standard deviation ( n = 6). &&& p < 0.001 versus sham group; * p < 0.05, ** p < 0.01, *** p < 0.001 versus LPS group; ## p < 0.01, ### p < 0.001 versus LPS + Olaparib (100 mg/kg) group.

Article Snippet: In the combination group, compound C (20 mg/kg; 99.10% purity; HY‐13418A, MedChemExpress) was co‐administered with Olaparib (100 mg/kg) 1 h after LPS injection.

Techniques: Staining, Control, Immunohistochemical staining, Activity Assay, Tissue, Standard Deviation

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Olaparib Blocks Oxidative Stress‐Dependent NLRP3 Inflammasome in Pediatric Acute Lung Injury by Metabolic Reprogramming via AMPK ‐Driven Autophagic Flux

doi: 10.1002/kjm2.70275

Figure Lengend Snippet: Olaparib reduces lung barrier dysfunction and apoptosis in ALI mice. (A–B) Protein expression levels of ZO‐1, claudin‐5, and VE‐cadherin in lung tissues from control, LPS, LPS + Olaparib (50 mg/kg), LPS + Olaparib (100 mg/kg), and LPS + Olaparib (100 mg/kg) + compound C (20 mg/kg) groups were analyzed by Western blot. (C–D) TUNEL staining and quantification of TUNEL‐positive area in lung tissues. (E–F) Western blot analysis of Bax and cleaved caspase‐3 protein expression in lung tissues. Data are presented as mean ± standard deviation ( n = 6). &&& p < 0.001 versus sham group; ** p < 0.01, *** p < 0.001 versus LPS group; ### p < 0.001 versus LPS + Olaparib (100 mg/kg) group.

Article Snippet: In the combination group, compound C (20 mg/kg; 99.10% purity; HY‐13418A, MedChemExpress) was co‐administered with Olaparib (100 mg/kg) 1 h after LPS injection.

Techniques: Expressing, Control, Western Blot, TUNEL Assay, Staining, Analysis, Standard Deviation

Journal: Frontiers in Aging Neuroscience

Article Title: Clemastine alleviates Parkinson’s disease pathology by activating the AMPK/mTOR pathway and enhancing autophagic flux

doi: 10.3389/fnagi.2026.1915475

Figure Lengend Snippet: CLE reduces α-synuclein protein levels and improves cell survival, effects inhibited by dorsomorphin (DOR). (A) Western blot (WB) and (B) The quantification of α-synuclein (α-syn) protein levels in PC12 cells from CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (C) Cell survival rate in CTRL, MPP + , and MPP + +CLE groups. (D, G) The WB analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in CTRL, DOR, CLE, and DOR+CLE groups. (E, F, H) The Quantification of the indicated protein ratios. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01 *** P < 0.001; ns, not significant. CLE, clemastine; α-syn, α-synuclein; CTRL, control; WB, western blot; DOR, dorsomorphin; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.

Article Snippet: The CTRL group was treated with normal saline (NS; volume equal to that of MPP + ), while the PD model group was treated with MPP + at a final concentration of 1 mM for 24 h. Cells in both CTRL and MPP + groups were subsequently treated with CLE, Baf-A1 (Selleck), or dorsomorphin (DOR; Selleck) according to the experimental design.

Techniques: Western Blot, Control, Analysis, Comparison

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Human iPSC-derived 2D cortical neurons and 3D cortical organoids show fetal brain proteomic similarity and distinct phosphoproteomic profiles

doi: 10.3389/fbioe.2026.1932407

Figure Lengend Snippet: Generation of human brain cortical organoids (COs) and functional validation using MEA recording. (a) Schematic workflow for the generation of human COs from human pluripotent stem cells. Cells were subjected to neural induction, neural progenitor cell expansion, and cortical neuronal differentiation using defined media conditions, extracellular matrix substrates, and stage-specific supplements. (b) Schematic illustration of a CO positioned on the electrodes of a microelectrode array (MEA) plate for extracellular electrophysiological recording. (c) Representative image showing a cortical organoid placed on the MEA electrode field. (d) Representative viability image demonstrating that the CO remained attached to the electrode area during recording. (e) Representative MEA raster plot showing spontaneous and evoked neuronal activity following NMDA application, indicating functional excitatory responsiveness of CO. (f) Representative MEA raster plots before, during, and after tetrodotoxin (TTX) treatment. TTX completely inhibited neuronal firing activity, and activity was restored after washout. (g) Quantification of weighted mean firing rate across conditions. Data represent mean ± SEM from individual organoids (n = 6). A repeated-measures one-way ANOVA was used to compare the paired measurements.

Article Snippet: Next day, the cells were 90%–100% confluent and differentiation was initiated by changing medium to Neurobasal medium (DMEM/F12, Neurobasal, 1X B-27 minus vitamin A, 1X N2 supplement, 1X L-Glutamine, 1X Non-essential amino acids (NEAA), 50 μM β-mercapto-ethanol, 0.4X Penicillin/streptomycin) supplemented with 10 μM SB431542 (Tocris, cat# 1614) and 2 μM Dorsomorphin (Tocris, cat# 3093) for 12 days.

Techniques: Organoids, Functional Assay, Biomarker Discovery, Microelectrode Array, Activity Assay

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: Protective effect of rMyonectin against LPS-induced apoptosis in cardiomyocytes is mediated by AMPK activation. (A) Western blot analysis and semi-quantification of p-AMPK and AMPK in myocardial tissue. (B) Western blot analysis and semi-quantification of p-AMPK and AMPK in NMCMs. (C) Expression of p-AMPK and AMPK in NMCMs after CC treatment. (D) Intracellular LDH activity in NMCMs. (E) The percentage of apoptotic cells detected using flow cytometry. (F) Apoptosis was assessed using flow cytometry after double labeling with Annexin V-FITC and PI. (G) Western blot analysis and semi-quantification of cleaved caspase-3, caspase-3, Bax and Bcl-2 protein expression in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; AMPK, AMP-activated protein kinase; CC, Compound C; LDH, lactate dehydrogenase; LPS, lipopolysaccharide; PI, propidium iodide; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Activation Assay, Western Blot, Expressing, Activity Assay, Flow Cytometry, Labeling, Recombinant

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: Protective effect of rMyonectin against LPS-induced mitochondrial dysfunction in cardiomyocytes is mediated by AMPK activation. (A) The ATP content in NMCMs. (B) Relative OCR. (C) Detection of the activities of mitochondrial respiratory chain complexes I and III. (D) Analysis of MMP using JC-1 staining. Scale bar, 50 μ m. (E) Western blot analysis and semi-quantification of PGC-1α, NRF1, TFAM, OPA1, Mfn2, p-Drp1 at Ser616, and Drp1 in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; OCR, oxygen consumption rate; MMP, mitochondrial membrane potential; PGC-1α, peroxisome proliferator-activated receptor γ co-activator-1 α; NRF1, nuclear respiratory factor 1; TFAM, mitochondrial transcription factor A; Mfn2, mitofusin 2; OPA1, optic atrophy 1; Drp1, dynamin-related protein 1; LPS, lipopolysaccharide; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Activation Assay, Staining, Western Blot, Recombinant, Membrane

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: AdipoR1 knockdown abolishes the protective effect of rMyonectin against LPS-induced apoptosis in cardiomyocytes. (A) Western blot analysis and semi-quantification of AdipoR1 in myocardial tissue. (B) Western blot analysis and semi-quantification of AdipoR1 in NMCMs. (C) Representative western blots showing the expression of AdipoR1, p-AMPK and AMPK in NMCMs following AdipoR1 knockdown. (D) Semi-quantification of AdipoR1, p-AMPK and AMPK protein levels in NMCMs following AdipoR1 knockdown. (E) Intracellular LDH activity in NMCMs. (F) The percentage of apoptotic cells detected using flow cytometry. (G) Apoptosis was assessed using flow cytometry after double labeling with Annexin V-FITC and PI. (H) Western blot analysis and semi-quantification of cleaved caspase-3, caspase-3, Bax, and Bcl-2 protein expression in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; AdipoR1, adiponectin receptor 1; AMPK, AMP-activated protein kinase; LDH, lactate dehydrogenase; si, small interfering RNA; NC, negative control; siAdipoR1, siRNA targeting AdipoR1; LPS, lipopolysaccharide; p-, phosphorylated; PI, propidium iodide.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Knockdown, Western Blot, Expressing, Activity Assay, Flow Cytometry, Labeling, Recombinant, Small Interfering RNA, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: AdipoR1 knockdown abolishes the protective effect of rMyonectin against LPS-induced mitochondrial dysfunction in cardiomyocytes. (A) The ATP content in NMCMs. (B) Relative OCR. (C) Detection of the activities of mitochondrial respiratory chain complexes I and III. (D) Analysis of MMP using JC-1 staining. Scale bar, 50 μ m. (E) Western blot analysis and semi-quantification of PGC-1α, NRF1, TFAM, OPA1, Mfn2, p-Drp1 at Ser616 and Drp1 in NMCMs. The data are presented as mean±SEM. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. rMyonectin, recombinant myonectin; NMCMs, neonatal mouse cardiomyocytes; AdipoR1, adiponectin receptor 1; AMPK, AMP-activated protein kinase; OCR, oxygen consumption rate; MMP, mitochondrial membrane potential; PGC-1α, peroxisome proliferator-activated receptor γ co-activator-1 α; NRF1, nuclear respiratory factor 1; TFAM, mitochondrial transcription factor A; Mfn2, mitofusin 2; OPA1, optic atrophy 1; Drp1, dynamin-related protein 1; si, small interfering RNA; NC, negative control; siAdipoR1, siRNA targeting AdipoR1; LPS, lipopolysaccharide; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Knockdown, Staining, Western Blot, Recombinant, Membrane, Small Interfering RNA, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Recombinant myonectin ameliorates sepsis-induced cardiomyopathy by alleviating mitochondrial dysfunction via the AdipoR1/AMPK pathway

doi: 10.3892/ijmm.2026.5900

Figure Lengend Snippet: Molecular mechanism by which rMyonectin ameliorates SIC. rMyonectin ameliorates SIC by alleviating mitochondrial dysfunction and inhibiting cardiomyocyte apoptosis via activation of the AdipoR1/AMPK pathway. rMyonectin, recombinant myonectin; SIC, sepsis-induced cardiomyopathy; OMM, outer mitochondrial membranes; IMM, inner mitochondrial membranes; AdipoR1, adiponectin receptor 1; AMPK, AMP-activated protein kinase; PGC-1α, peroxisome proliferator-activated receptor γ co-activator-1 α; NRF1, nuclear respiratory factor 1; TFAM, mitochondrial transcription factor A; Mfn2, mitofusin 2; OPA1, optic atrophy 1; Drp1, dynamin-related protein 1; I, mitochondrial respiratory chain complex I; III, mitochondrial respiratory chain complex III; p-, phosphorylated.

Article Snippet: NMCMs were pretreated for 2 h with rMyonectin or with the AMPK inhibitor Compound C (CC; 10 μ M; cat. no. HY-13418A; MedChemExpress) ( , ), followed by stimulation with 10 μ g/ml LPS for 24 h ( ).

Techniques: Activation Assay, Recombinant